Lupus (2010) 19, 496–546
http://lup.sagepub.com
ABSTRACTS
Abstract presentations Wednesday April 14th, 2010
A001
Induction of pathogenic antiphospholipid antibodies in vivo are dependent
on expression of MHC-II genes
Papalardo E1, Romay-Penabad Z1, Christadoss P1, Pierangeli S1
1 1
University of Texas Medical Branch, Galveston, TX
Background: Antiphospholipid Syndrome (APS) is an autoimmune
disease characterized by the presence of antiphospholipid (aPL)/anti-
b2Glycoprotein I (anti-b2GPI) antibodies (Abs) in association with
recurrent thrombosis and/or pregnancy losses. Pathogenic aPL auto-
antibodies can be generated by immunization of mice with human
b2GPI and a molecular mimicry mechanism has been proposed.
However, predisposing genetic factors have not been fully investigated
in APS. Material and Methods: Here, we studied the influence of
MHC-II and the expression of certain human haplotypes (DQ6,
DQ8, and DR4) on the production of pathogenic of aPL Abs in
mice. MHC-II deficient (-/-), DQ6, DQ8 and DR4 mouse MHC-II-
deficient-transgenic mice or their corresponding wild-type (WT) ani-
mals were immunized with human b2GPI or with control protein oval-
bumin (OA). The production of aCL and anti-b2GPI abs in the sera of
the mice was monitored weekly. Thrombus formation, tissue factor
(TF) activity in mouse peritoneal macrophages and in carotid artery
homogenates was examined in the mice. Results: WT mice produced
significantly higher titers of aCL and anti-b2GPI abs in mice immu-
nized with b2GPI when compared to controls and those correlated with
enhanced thrombus formation and TF activity. MHC(-/-) immunized
with b2GPI did not produce aCL or anti-b2GPI Abs and mean throm-
bus sizes as well as TF activities were significantly diminished, when
compared to WT animals. Pathogenicity of aPL Abs was fully
re-established in MHC (-/-) mice expressing DR4, DQ8 haplotype
and partially restored in animals expressing human DQ6.
aCL Titer a2 GPI Titer
(OD units) (OD units)
Mice/Treatment Mean  SEM Mean  SEM
WT þ OA *0.17  0.20 *0.31  0.03
WT þ b2 GPI 1.77  0.040 2.81  0.04
MHC (-/-) þ OA 0.13  0.03 0.29  0.03
MHC (-/-) þ b2 GPI *0.17  0.06 *0.433  0.28
*Statistically significant different from WT þ b2GPI group.
Conclusions: The data shows for the first time that generation of patho-
genic aPL Abs in vivo depend on expression of MHC-II genes and that
certain haplotypes may protect or make individuals more susceptible
to the development of APS.
Disclosure: Nothing to disclose.
! The Author(s), 2010. Reprints and permissions: http://www.sagepub.co.uk/journalsPermissions.nav
A002
Induction of anti-b2-GPI antibodies by Streptococcus pyogenes surface
protein H
van Os G1,2, Herwald H3, Derksen R1, Meijers J2, de Groot P1
UMC Utrecht, The Netherlands; 2AMC, The Netherlands;
3
Lund university, Sweden
Introduction: The antiphospholipid syndrome (APS) is characterized
by the presence of antibodies to b2-glycoprotein I (b2-GPI). These
auto-antibodies recognise a cryptic epitope within Domain I of
b2-GPI. We have found that b2-GPI interacts with Streptococcus pyo-
genes. To study if an infection with S. pyogenes can induce APS, mice
were injected with four S. pyogenes surface proteins to determine their
APS related pathogenic potential. Methods and Materials: BALB/c
mice (n¼8) were injected intraperitoneally with 25 mg M1 protein,
Protein H, Streptococcus collagen like protein A and B (SclA and
SclB) or human serum albumin in saline for a total of 6 boosts.
Plasmas were collected from mice and then every 4 weeks the proteins
were injected. Plasmas were collected two weeks after each protein
injection. Results: IgGs isolated from APS patients were not able to
recognize b2-GPI in solution. However, only after the addition of
Protein H the patient antibodies recognised soluble b2-GPI. Injection
of bacterial proteins did not lead to an apparent clinical phenotype.
Although, mice injected with M1 protein, SclA and SclB developed
antibodies directed against the injected protein, these mice did not
develop antibodies towards b2-GPI. In contrast, all 8 mice injected
with Protein H showed besides anti-Protein H antibodies also antibo-
dies directed against mouse b2-GPI from boost 2 on. Domain specific
ELISAs showed that all 8 mice developed antibodies directed only
against Domain I of b2-GPI.
Plasma from mice injected with Protein H showed a prolongation
of the APTT. IgGs were isolated from pooled plasma after the last
Thrombosis
boost from mice injected with Protein H. Addition of 10 mg/ml IgG to
(m2) human plasma prolonged the PTT-LA coagulation time from 39 to
Mean  SD 61 seconds. No prolongation was observed with an APTT activated by
Actin FS, a lupus anticoagulant insensitive reagent. Discussion/
*615  169 Conclusion: Our study showed that Protein H from S. pyogenes induces
2978  501 a conformational change within b2-GPI, that leads to the exposure of a
305  106 cryptic epitope on Domain I, the development of anti-b2-GPI antibo-
*767  145 dies and lupus anticoagulant activity. These data show that repeated
exposure of a cryptic epitope within b2-GPI is sufficient to induce an
autoimmune response.
Disclosure: Nothing to disclose.
10.1177/0961203310365200

A003
Influenza vaccination can induce new onset anticardiolipins but not
b2-glycoprotein antibodies among patients with systemic lupus
erythematosus
Vista E1, Crowe S1, Dedeke A1, Anderson J1, Thompson L1,2, Air G2,
James J1,2
1
Oklahoma Medical Research Foundation, Oklahoma City, OK;
2
University of Oklahoma Health Sciences Center, Oklahoma City, OK
Purpose: Report the association between antiphospholipid antibodies
and influenza vaccine among patients with systemic lupus erythemato-
sus (SLE). Methods: Sera of 80 patients fulfilling the SLE American
College of Rheumatology criteria and 88 matched healthy controls
who received seasonal influenza vaccine were tested by ELISA for
anticardiolipin (aCL) IgG at the time of vaccination and after 2, 6
and 12 weeks. Those with reactivity to aCL IgG were further tested
for B2-glycoprotein Independent (b2GPI) IgG antibodies. Vaccine
responses were ranked according to an overall anti-influenza antibody
response index. Results: More SLE patients (10 of 80) compared to
controls (2 of 88) developed new aCL reactivity (p 0.0324 by two-tailed
Fisher’s exact test). No new anti-b2GPI were detected in patients or
controls. Baseline median number of SLE criterion is 6 (95% CI 5.45
to 7). Differences were seen in vaccine response among aCL positive
individuals (p¼0.7968). Conclusion: This study shows transient
increases in aCLs but not anti-b2GPI after influenza vaccination sup-
porting the possibility that these changes in autoantibody levels may
not be clinically important for increased thrombosis risk.
Disclosure: This presentation was made possible by a full grant from
the National Institute of Allergy and Infectious Diseases
(HHSN266200500026C).
A004
Genetic variants associated with antiphospholipid antibodies
Wen YY1, Thiagarajan P1, Gibbs R1, Arnett F2
1
Baylor College of Medicine, Houston, Texas; 2The University of Texas
Health Science Center, Houston, Texas
Antiphospholipid syndrome (APS) is an acquired autoimmune disor-
der characterized by thrombosis, or recurrent pregnancy morbidity in
the presence of antiphospholipid antibodies. Given that genetic factors
have been suggested to influence the etiology of APS, there has been
great effort in determining the APS associated loci. The aim of the
present study was to investigate whether genes involved in homeostatic
pathways are associated with development of APS in patients with
lupus or allied autoimmune disorders. We selected 89 patients (all
North Americans of European descent) with positive antiphospholipid
antibodies or other autoimmune disorders. For a normal population
control, we selected 94 ethnically matched HapMap CEU samples. To
detect single nucleotide polymorphisms (SNPs) between patients and
normal controls, we studied 129 genes involved in the coagulation
pathway. Target regions were first enriched by Capture NimbleGen
solid surface arrays, followed by 454 GS FLX Titanium sequencing.
We further confirmed 27 SNPs identified from the 454 sequence by
Sanger sequencing. Minor allele frequency was calculated, permitting
identification of 3 loci of genetic susceptibility of APS. A genetic var-
iant in FCGR2B intron region (SNP rs2045572, p ¼ 7.22*106) was
detected in 10% of the APS patients with antiphospholipid antibodies,
and in none of control samples. However, this genetic variant is a SNP
in HapMap YRI population with minor allele frequency of 0.133. A
SNP (rs28395877, p ¼ 0.015) in PLCB3 exon region was associated
with the susceptibility of APS. Homozygous AA variant of rs28395877
increases disease risk more than the homozygous GG allele in the
Abstracts
497
normal population. In addition, a missense change (rs8386, p ¼
0.007) in the coding region in GNAS was identified as another APS
susceptibility locus in our study. We identified polymorphisms in
PLCB, guanine GNAS, and SNP rs2045572 in the FCGR2B intronic
region to be associated with the development of antiphospholipid anti-
bodies in the Caucasian population.
Disclosure: Nothing to disclose.
A005
Oxidation of antibodies as a trigger for autoimmunity
Omersel J1, Cucnik S1, Avbersek Luznik I2, Kveder T1, Rozman B1,
Bozic B1,3
1
University Medical Centre, Ljubljana, Slovenia; 2General Hospital
Jesenice, Jesenice, Slovenia; 3University of Ljubljana, Ljubljana,
Slovenia
Objectives: The aim of our study was to determine the binding capacity
alterations of oxidized IgGs and IgMs, to evaluate a subsequent bind-
ing to some autoantigens, and to evaluate their effects on released
cytokines in endothelial cells. Methods and Materials: IgG and IgMs
were purified from healthy blood donors and exposed to direct current
(15-300 sec) at different electromotive force (2-20 mA, 1.5–9V) in PBS
with or without retinol, ascorbic acid or Igs depleted serum. Altered
immunoreactivity was tested by anti-b2GPI, aCL, aCCP, aPR3 and
RF ELISAs, IIF and CIE. IL-6 mRNA and protein levels were deter-
mined by RT-PCR and ELISA. Results: Alterations in immunoreac-
tivity of oxidized IgGs were observed with all voltages used, with the
maximum binding to autoantigens after 15-60 s of oxidation. Addition
of retinol, vitamin C or IgG depleted serum during electro-oxidation
decelerated changes in specificity of natural IgG. Treatment of
HUVEC with an oxidized IgGs resulted in changed cell morphology
and in an increase in IL-6. Clinically important increase in immunor-
eactivity of IgM was induced with 6.5 mA, after 30-60 sec, giving low
to medium titers in aCL, and low positive titers in anti-b2GPI ELISA,
but negative on RF. Conclusions: Electro- or chemically induced oxi-
dation of IgMs and IgGs from healthy persons progresses to auto-
immunoreactivity. Due to a complex structure of IgM susceptibility
to the oxidative treatment was expectedly different from the deter-
mined IgGs’ oxidation pattern. Oxidative changes of IgG and IgM
support a hypothesis that oxidative stress could be a putative trigger
of autoimmunity.
Disclosure: The study has been partially financed by Ministry of
Higher Education, Science and Technology, Slovenia, Grant Nos:
P3-0314.
A006
Biodistribution of b2GPI in naive and immunized mice and in vivo pro-
thrombotic effect of an anti-b2GPI minibody isolated from human phase
display library
Tedesco F1
1
University of Trieste, Trieste, Italy
Background: b2GPI has been shown to bind to a variety of cells includ-
ing endothelial cells, platelets, monocytes and trophoblasts. Most of
the information has been obtained from in vitro and ex vivo studies.
The aim of our investigation was to evaluate the in vivo binding of
b2GPI to placenta and to the vascular endothelium prior and after
immunization, and also to assess the in vivo effect of a novel recombi-
nant human antibody to b2GPI. Methods: b2GPI purified from human
plasma was labeled with the fluorescent probe Cy 5.5 and injected iv
into female Balb-c mice either naive or immunized with human b2GPI.
Lupus
 Abstracts
498
The in vivo and ex vivo distribution of the labeled protein was evaluated
by time-domain eXplore Optix pre-clinical imager. Binding of b2GPI
to various organs was also checked by confocal analysis. The human
antibody to b2GPI was isolated as scFv from human phage display
library and further engineered to contain the hinge-CH2-CH3 domains
of human IgG1. Results: The labeled protein injected into naive mice
was cleared from the circulation within 48 hours mostly by the liver
and was eventually excreted in the urine. Ex vivo analysis performed 7
days after b2GPI injection confirmed the accumulation of the protein
in the liver. Interestingly, some staining was observed in the uterus.
The protein distribution was essentially similar in immunized animals
that produced high titer antibodies to human b2GPI non cross-reactive
with the mouse protein. The labeled protein injected into pregnant
naive mice was cleared mostly by the liver and placenta and no
major differences were observed in immunized pregnant animals
except that in this group of mice the presence of antibodies was asso-
ciated with an increased fetal resorption rate and local deposition of C3
and C9. We failed to see vascular thrombosis in any of the organs
examined suggesting that b2GPI does not bind to the endothelium
unless the endothelial cells are primed by activating stimuli. This find-
ing was confirmed using a human anti-b2GPI minibody, that was able
to induce thrombosis via complement activation only in mice primed
with LPS.
The results suggest that b2GPI interacts directly with the feto-pla-
cental unit while requiring prior cell activation to bind to the
endothelium.
Disclosure: Nothing to disclose.
A007
Lupus susceptible gene polymorphisms in patients with antiphospholipid
syndrome in Japanese population
Horita T1, Nakagawa H1, OKu K1, Kataoka H1, Yasuda S1, Atsumi T1,
Koike T1
1
Hokkaido University, Sapporo, Hokkaido, Japan
Purpose: Genome wide association studies in patients with SLE in
Caucasian population revealed several novel lupus susceptible genes
in addition to previously reported genes such as HLA, interferon reg-
ulatory factor 5 (IRF5), signal transducer and activator of transcrip-
tion 4 (STAT4) and so forth. In this study, we investigated the possible
association between these novel lupus susceptible gene polymorphisms
and APS in Japanese population. Method: Genomic DNA samples
were extracted from 341 SLE patients, 90 APS patients (41 primary
APS and 49 secondary APS complicated with SLE) and 428 ethnically
matched healthy controls. Single nucleotide polymorphisms (SNPs) of
B-cell scaffold protein with ankyrin repeats 1 (BANK1), B lymphocyte
specific tyrosine kinase (BLK), Phox homology domain containing
serine/threonine kinase (PXK), tumor necrosis factor alpha induced
protein 3 (TNFAIP3), tumor necrosis factor ligand superfamily
member 4 (TNFSF4) and rs10798269 in 1q25.1 region were genotyped
using TaqMan Genotyping Assay kit. Allele frequencies in each group
were compared using chi-square test and the related risk was approxi-
mated by the odds ratios. Results: BANK1 (rs10516487), BLK
(13277113) and rs10798269 in 1q25.1 region were associated with not
only SLE but also APS in Japanese population. TNFSF4 (rs844644)
association were found only in APS population whereas TNFAIP3
(rs2230926) association was found only in SLE. PXK (rs6445975)
was associated with neither SLE nor APS in Japanese population.
Conclusion: Our results suggest that APS and SLE, in part, share a
common genetic background.
Disclosure: Nothing to disclose.
Lupus
A008
HLA-DRB1 *0701 is associated with thrombosis but not
antiphospholipid antibody (aPL) production in systemic lupus
erythematosus (SLE)
Kaiser R1, Chung S1, Noble J2, Seldin M3, Taylor K1, Criswell L1
1
University of California, San Francisco, CA; 2Children’s Hospital
Oakland Research Institute, Oakland, CA; 3University of California,
Davis, CA
Purpose: We investigated whether HLA-DRB1 alleles were associated
with anticardiolipin (aCL) antibodies, the lupus anticoagulant (LAC)
or thrombosis. Methods: 909 Caucasian SLE patients fulfilling ACR
criteria for SLE were studied. Patients completed questionnaires doc-
umenting phenotypic, demographic, and medical information (con-
firmed by medical record review) and were genotyped for DRB1
using a PCR-sequence specific oligonucleotide probe methodology.
Global V2 tests were used to determine whether the DRB1 locus was
associated with the outcomes of interest and the relative predisposi-
tional effects method was used to identify associated alleles. Allelic V2
tests and logistic regression were used to further investigate associa-
tions between alleles and thrombosis, adjusting for ancestry and other
covariates. Results: The most common DRB1 alleles were *0301
(22%), *1501 (19%) and *0701 (14%). Global V2 tests for associations
between DRB1 and aCL (n¼630) and LAC (n¼297) were not signifi-
cant (p¼0.58 and p¼0.18), however the global V2 for thrombosis
(n¼757) was significant (p¼0.03), with *0701 contributing the most
to the overall V2. In allelic analyses, the OR for thrombosis and
*0701 was 1.42 (p¼0.03). In a multivariate model adjusting for aPL,
gender, smoking history, disease duration, nephritis and percent
Northern European ancestry, *0701 was associated with thrombosis
(OR 1.89, p¼0.04) as were aCL, LAC, smoking history, disease dura-
tion, and hydroxychloroquine (HCQ) treatment (HCQ was protective).
Conclusion: Although the HLA-DRB1 locus is strongly associated with
SLE, associations with certain autoantibodies and subphenotypes are
not well-established. Our results indicate that *0701 is associated with
thrombosis but not with aPL in SLE.
Disclosure: Nothing to disclose.
A009
Oxidation of monoclonal antibodies unmasks antiphospholipid
autoantibodies
McIntyre J1, Page Faulk W1
1
St. Francis Hospital, Beech Grove, IN
A novel family of antibodies recognizes autoantigens subsequent to
physiological oxidation-reduction (redox) reactions caused by hemin,
a normal product of erythrocyte aging and hemoglobin degradation.
The evolutionary persistence of this redox-reactive autoantibody (R-
RAA) family in animals implies that it is important for immunological
homeostasis. Initial information on R-RAA was obtained by studying
human blood and later human polyclonal immunoglobulin (IVIg) frac-
tions. Recently we asked if redox reactions had similar effects upon
monoclonal antibodies (mAb) and discovered that they did. Redox
reactions consistently unmask mAb to show specificities to phospholi-
pids and DNA. Despite the alterations in binding specificity induced by
oxidation, the mAb continues to bind the original autoantigen.
Experiments using antibody Fab fragments continued to show auto-
antibody unmasking, thus eliminating the participation of the antibody
Fc fragment in the observed oxidative changes. These findings raise
concerns that areas of oxidative stress in vivo could initiate redox-
mediated unmasking of mAb which could cause many of the
 Abstracts

untoward/unintended side effects known to occur with clinically
approved mAb therapies.
Disclosure: Nothing to disclose.
A010
APS clinical manifestations are associated with SNPs of inflammatory
genes
Broggini V1, D’Amelio F1, Raschi E2, Grossi C2, Tincani A4, Roubey R3,
Meroni PL1, Borghi MO1
1
Univ MI, Milan, Italy; 2Ist Auxologico It, Milan, Italy; 3Univ NC,
Chapel Hill, NC; 4Univ Brescia, Brescia, Italy
Background: There is sound evidence that aPL are pathogenic; how-
ever they display the thrombogenic effect in animal models only in the
presence of a second inflammatory hit. This finding fits well with the
clinical observation that aPL can be persistently positive but thrombo-
tic events occur only occasionally, frequently after an inflammatory
process. Animal models as well as preliminary studies on SNPs of
candidate genes of inflammatory mediators suggest that a proinflam-
matory phenotype may favour the clinical manifestations of APS. Aim:
to investigate the role of the host’s genetic background as a variable
critical for the clinical manifestations of the syndrome. Material and
Methods: 135 primary APS (PAPS) with thrombotic events, 270 NHS
and 200 aPL-negative patients with thrombosis (aPL-/thr) were
enrolled. An Illumina Golden Gate Assay for 768 SNPs in 62 genes
of inflammatory signaling pathways was performed. Statistics were
carried out with PLINK software while haplotype analysis with
Haploview4.1. Results: PAPS vs NHS analysis identified 31 significant
SNPs in 11 inflammation genes; particularly, 4 SNPs in nox3, 1 in
mapk8 and 2 in irak2 were strongly associated; the association
remained significant after FDR correction for nox3. Haplotype analy-
sis grouped 7 nox3 SNPs in one block in Linkage Disequilibrium and
the haplotype consistent with individual SNP analysis was statistically
significant. PAPS vs aPL-/thr analysis displayed similar results and no
remarkable data emerged when comparing aPL-/thr vs NHS. Analysis
of an additional 160 aPLþ asymptomatic subjects is under evaluation.
Conclusions: SNPs of proinflammatory genes are associated to thromb-
sis in APS. Nox3, MAPK8, IRAK2 play a role in TLR-mediated
signalling pathway. Our results may support the link between the
innate immunity responses and the clinical manifestations of APS.
499
controls were compared and correlations between the presence of
aPL with clinical manifestations were performed. Results: APS patients
and controls showed no differences in prevalence of both codons.
Heterozygous genotypes of Val247Leu polymorphism affected 18
patients and 7 cases of homozygous L/L genotype. Heterozygous
T/S genotype was showed in 14 patients and 10 controls and no
cases of homozygous (S/S) were found. No significant correlation
was found between the presence of a particular polymorphism and
the presence of aPL antibodies. Clinical variables as type of APS,
type of thrombosis, neurological and cutaneous manifestations as
well heart valve disease did not correlate with studied polymorphisms.
Conclusion: Polymorphisms at codons 247 and 316 of b2GPI gene are
similarly expressed in Spanish APS patients and the general popula-
tion. b2GPI gene polymorphisms seem not to be involved in the ser-
ological and clinical features of the disease in our population.
Disclosure: The authors have no potential conflict of interest.
A012
Association between CD36 single nucleotide polymorphism and
antiphospholipid syndrome
Kato M1, Horita T1, Atsumi T1, Fujieda Y1, Otomo K1, Yasuda S1,
Koike T1
1
Hokkaido University Graduate School of Medicine, Sapporo,
Hokkaido, Japan
Purpose: CD36, known as a scavenger receptor, is a transmembrane
glycoprotein expressed on monocytes, platelets and capillary endothe-
lial cells. CD36 recognizes multiple ligands, including phosphatidyl
serine, and is a mediator of both atherogenesis and thrombosis. The
purpose of this study is to investigate the association between CD36
gene polymorphisms and antiphospholipid syndrome (APS). Methods:
This study comprised 795 Japanese: 39 patients with primary APS, 69
with systemic lupus erythematosus (SLE) complicated with APS, 265
with SLE in the absence of APS, and 422 healthy subjects. Two follow-
ing CD36 gene polymorphisms were investigated in this population
using the TaqMan PCR genotyping method, rs3765187 a common
variation linked to CD36 deficiency, and rs3765187 on the 50 untrans-
lated region. Result: The minor allele frequency of rs3765187, pre-
sented in the table, was less frequent in APS patients than in healthy
subjects.

Disclosure: Nothing to disclose.

Odds
Minor allele ratio
A011 frequency p value (95% CI)

Healthy subjects 10.2% (43/422) - -

Lack of association between b2-glycoprotein I gene polymorphisms with
(n ¼ 422)
antiphospholipid antibodies in Spanish APS patients
All APS (n ¼ 108) 2.8% (3/108) 0.024 0.25 (0.08 to 0.83)
Castro-Marrero J1, Pardos-Gea J1, Balada E1, Corte´s-Hernández J1,
All SLE (n ¼ 334) 6.9% (23/334) 0.11 0.65 (0.38 to 1.11)
Vilardell-Tarre´s M1, Ordi-Ros J1
1 Primary APS (n ¼ 39) 2.6% (1/39) 0.15 0.23 (0.03 to 1.73)
Vall d’Hebron University Hospital Research Institute, Barcelona, Spain
SLE þ APS (n ¼ 69) 2.9% (2/69) 0.085 0.26 (0.06 to 1.11)
SLE/non-APS (n ¼ 265) 7.9% (21/265) 0.32 0.76 (0.44 to 1.31)
APS is a disease characterized by antiphospholipid antibodies (aPL)
and at least one clinical manifestation, the most common being venous
and arterial thrombosis and recurrent fetal loss. Objective: To deter-
There was no significant difference in the allele frequency of rs1049654
mine the presence of polymorphisms at codons 247 (Val247Leu) and
among those groups. Conclusion: The single nucleotide polymorphism
316 (Trp316Ser) of b2GPI gene in Spanish APS patients and healthy
linked to CD 36 deficiency was less frequent in APS patients, suggest-
controls and their possible correlation with the development of aPL
ing that impaired scavenger receptor function correlates with APS-
and associated clinical manifestations. Methods: PCR amplified both
resistant. This is a first report to show the link between CD36 gene
codons from b2GPI gene and the presence of polymorphisms was
and APS.
detected by PCR-RFLP. Antiphospholipid antibodies were detected
by ELISA. Allelles and genotype frequencies in APS patients and Disclosure: Nothing to disclose.

Lupus
 Abstracts

500
A013 40% in platelet adhesion compared to non-reduced b2GPI when vWF
was activated with ristocetin. Platelet adhesion was partially inhibited
Proteomic analysis shows different patterns of protein expression in by addition of the TRX-R inhibitor DNCB implying that reduction of
monocytes exposed to IgG from patients with different manifestations b2GPI by TRX-1 was responsible partially for the adhesion of the
of APS platelets. (figure 2) B). Reduced b2GPI resulted in a 42% increase in
Lambrianides A1, Heywood W1, Mills K1, Bell K1, Latchman D1, platelet adhesion to endothelial cells compared to non-reduced b2GPI
Isenberg D1, Pierangeli S2, Rahman A1, Giles I1 or HSA(p¼0.02) (figure 3).
1
University College London, London, United Kingdom; 2University of
Texas, Galveston, TX

A major mechanism of hypercoagulability in the Antiphospholipid

Syndrome (APS) is increased expression of tissue factor on monocytes
activated by antiphospholipid antibodies (aPL). Previously we found
differences in activation of p38MAPK and NFkB signaling pathways
in monocytes exposed to purified IgG from different clinical subgroups
of APS - patients with vascular thrombosis (VT) but no pregnancy
morbidity (PM) and those with PM but no VT. Using proteomic ana-
lysis, our aim was to identify other proteins that are differentially
affected. Polyclonal IgG was protein G purified from 14 patients
with APS (7 VTþ/PM- and 7 VT-/PMþ) and 7 aPL-negative healthy
controls. A single sample for each of the 3 groups was produced by
pooling the IgG from the 7 subjects in that group and determined to be
endotoxin-free. U937 cells were treated with 100mg/ml IgG from each
pooled sample for 6 hours and lysates examined by fluorescence 2D
difference gel electrophoresis (DIGE). The analysis revealed that 20
proteins were up-regulated in both APS samples compared to healthy
controls by between 2 and 3.4-fold. Five proteins were down-regulated
in both APS samples compared to controls by between 2 and 2.5-fold.
There were also clear differences in the expression/modification of
proteins in monocytes treated with IgG from patients with different Conclusions: b2GPI is involved in thiol exchange reactions with Trx-1
clinical manifestations of the APS. We identified 7 proteins that were and PDI, and through this mechanism can enhance the binding of
up-regulated between 2 and 3.1-fold and 3 proteins that were down- platelets to vWF and endothelial cells.
regulated between 2 and 2.2-fold in patients with VTþ/PM- compared Disclosure: No response indicated.
to patients with VT-/PMþ. Further analysis is ongoing to determine
the identity of these proteins. Identification of these proteins that are
significantly altered will provide further insight into pathogenic A015
mechanisms of the APS and may provide future alternative therapeutic
targets. b2glycoprotein-I(b2GPI)-related novel peptide attenuates anti-b2GPI-
activated -HUVEC via GSK3 inhibition and reduces experimental APS
Disclosure: Nothing to disclose.
Blank M1, Prof., Shoenfeld Y1, Eisenstein M2, Fridkin M2, Baraam L1,
Heldman Y2, Katchalski-Katzir E2
1
A014 Sheba Medical Center, Ramat-Gan, Tel-Hashomer, Israel;
2
The Weizmann Institute of Sciences, Rehovot, Rehovot, Israel
Participation of reduced beta 2 glycoprotein I by thioredoxin-1 (TRX-1)
in the regulation of platelet adhesion through binding of von Willebrand Background and Aim: In-vitro and in-vivo studied revealed that high
factor in a thiol dependent manner titers of anti-phospholipid/ b2GPI-binding-antibodies(Abs) are major
Qi M1, Freda Passam1, Qi JC1, Giannakopoulos B1, Krilis S1 contributors to activation of platelets and endothelial-cells (ECs)
1
University of New South Wales, St. George Hospital, Sydney, Australia in-vitro, as well as induction of fetal loss and thromboses in-vivo.
Patients with APS harbor diverse clusters of circulating anti-b2GPI
Abstract purpose: We demonstrate the novel finding that reduced Abs which targets different epitopes on the b2GPI. Several anti-
b2GPI by TRX-1 increases platelet adhesion to activated vWF and b2GPI-neutralizing synthetic peptides were identified. Our approach
endothelial cells. Methods: A). Platelet adhesion to b2GPI coated was to find a novel peptide for therapy, based on b2GPI –HUVEC-
microtitre wells in the presence of vWF. Plates were coated with binding-site (phospholipids and membrane), named EMBI. Results:
non-reduced or reduced rb2GPI (by activated TRX-1) and incubated EMBI inhibited differentially the binding of anti-b2GPI Abs from 7
with activated vWF. Platelets with or without activated TRX-1 or APS patients to ECs, up to 89% (9.3% inhibition by scEMBI)
DNCB (TRX-1 inhibitor) were aliquoted into the corresponding P<0.001. Furthermore, EMBI inhibited E-selectin expression 7 to 9.6
wells and incubated, platelets that had adhered to the wells were visua- times more than did scEMBI (p<0.002). The expression of tissue factor
lised with microscope and quantitated. B). Platelet adhesion to human (TF) was decreased up to 92% upon treatment with EMBI in compar-
umbilical endothelial cells (HUVECs). HUVECs were cultured on ison to 7% inhibition with scEMBI. The data were supported by real-
plates until confluence. b2GPI or HSA alone (or treated with activated time-PCR. Employing phospho-MAPK-arrays indicated an enhanced
TRX- 1) were added to the endothelial cells. Washed platelets were expression of phosphorylated JNK1/2, p38, and HSP27 and decreased
labeled with Calcein-AM and added to the endothelial cells containing expression of phospho-glycogen synthase kinase-3 (pGSK3).
the different b2GPI or HSA preparations. After incubation and wash- Knocking down the GSK3b expression by siRNA-GSK3b reduced
ing, the fluorescence in the wells was read by a fluorescence plate the TF expression by anti-b2GPI-activated-HUVEC from 2 APS
reader. Results: A). Reduced b2GPI caused a significant increase by patients. In-vivo, EMBI significantly decreased the percentage of fetal

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loss in naı̈ve mice infused with anti-b2GPI Abs. Conclusion: EMBI may
serve as a potential novel candidate peptide, to treat patients with APS.
Disclosure: Nothing to disclose.
A016
Histidine-rich-glycoprotein: a protective molecule in anti-b2GPI
mediated endothelial cell induced procoagulation state
Blank M, MD1, Buccarelli S2, Cervera R2, Espinosa G2, Altman A1,
Praprotnik S3, Shoenfeld Y1
1
Sheba Medical Center, Ramat-Gan, Tel-Hashomer, Israel;
2
Hospital Clinic, Barcelona Catalonia, Spain; 3University Clinical
Center, Ljublijana, Slovenia
Background and Aims: Histidine-rich glycoprotein (HRG) is an
a2-glycoprotein found in mammalian plasma (150 mg/ml). HRG
binds to cell surfaces and numerous ligands such as plasminogen, fibri-
nogen, thrombospondin, C1q, heparin, glycosaminoglycans (GAGs),
suggesting that it may act as an adaptor protein either by targeting
ligands to cell surfaces or by cross-linking soluble ligands. HRG has a
role in inflammation, during tumor metastasis and angiogenesis. HRG
and b2GPI are GAGs binding proteins. The interaction between
endothelial-cells, HRG, b2GPI and anti-b2GPI mediated endothelial
cell activation was analyzed. Methods: HRG was affinity purified from
healthy donor plasma on heparin and Nickel columns. Anti- b2GPI
Abs mediated endothelial cell activation (e.g. E-selectin, tissue factor
expression and IL-6 secretion) was analyzed by ELISA immunoblot
and real-time PCR. Results: Heparan sulfate is the predominant
human umbilical vein endothelial cell (HUVEC) surface ligand for
HRG since mammalian heparanase inhibit the HRG-endothelial cell
binding. HRG compete with b2GPI for endothelial cell binding by
86%. Therefore, HRG prevent the anti-b2GPI binding to b2GPI /
endothelial-cells ranging 75-90% P<0.001. HRG inhibits E-selectin
and tissue factor expression induced by anti-b2GPI Abs, depicted by
immunoblot and real-time PCR. Conclusions: Our in-vitro results raise
the posibility that HRG is a protective self molecule for anti-b2GPI
Abs mediated endothelial cell activation and can attenuate procoagu-
lation state.
Disclosure: Nothing to disclose.
A017
Two conformations of b2-Glycoprotein I
Ağar Ç1,2, van Os G2, Mörgelin M3, Sprenger R1, Marquart A1,
Urbanus R2, Derksen R2, de Groot P2, Meijers J1
1
Academic Medical Center, Amsterdam, The Netherlands;
2
University Medical Center, Utrecht, The Netherlands;
3
Lund University, Lund, Sweden
The antiphospholipid syndrome is defined by the presence of antipho-
spholipid antibodies in blood of patients with thrombosis or fetal loss.
There is ample evidence that b2-glycoprotein I (b2GPI) is the major
antigen for antiphospholipid antibodies. The auto-antibodies recognize
b2GPI when bound to anionic surfaces and not in solution, suggesting
that b2GPI bound to a surface has another conformation than b2GPI
in solution. Here we showed with three independent techniques that
b2GPI can exist in at least two different conformations, a circular
conformation and an ‘activated’ open conformation, identical to the
published crystal structure. We were able to convert the conformation
of b2GPI from the closed to the open conformation and back, by
changing pH and salt concentration. The closed, circular conformation
is maintained by interaction between the first and fifth domain of
b2GPI and is the major form of b2GPI in plasma. In the circular
Abstracts
501
conformation the epitope for the antibodies is shielded from recogni-
tion. After binding to anionic surfaces, b2GPI converts into the open
conformation. The cryptic epitope in the first domain becomes
exposed, enabling patient antibodies to bind and form an antibody-
b2GPI complex. We also demonstrate that the open conformation of
b2GPI prolonged the aPTT when added to normal plasma, with an
even further prolongation by addition of anti-b2GPI antibodies. The
conformational change of b2GPI and the influence of the auto-anti-
bodies may have important consequences for our understanding of the
antiphospholipid syndrome.
Disclosure: Nothing to disclose.
A018
Oxidative stress as a pathogenic link between aPL, thrombosis and
atherosclerosis development in antiphospholipid syndrome
López-Pedrera C1, Ruiz-Limón P1, Barbarroja N1, Rodrı´guez-Ariza A1,
Collantes-Este´vez E1, Velasco F1, Khamashta M2, Cuadrado MJ2
1
Reina Sofia Hospital-IMIBIC, Cordoba, Spain; 2St Thomas Hospital,
London, United Kingdom
The aim of this study was to investigate the association of oxidative
stress and mitochondrial membrane potential (MMP) in leucocytes
from 20 APS patients with the development of thrombosis and ather-
osclerosis. Increased expression of TF and PARs were found in mono-
cytes from APS patients, which also displayed higher plasma levels of
VEGF, tPA, sP-selectin, and MCP1. APS monocytes and neutrophils
showed increased peroxides and superoxide levels and more depolar-
ized mitochondria, while intracellular GSH was significantly decreased.
Healthy monocytes stimulated (6h) with APS, but not with control
IgG, also displayed increased peroxides and superoxide levels and
decreased MMP and GSH. Monocyte MMP and peroxides showed
significant negative and positive correlations, respectively, with TF
expression. aPL-IgG positively correlated with peroxides, TF, VEGF
and PAR2, and negatively with MMP and GSH. Carotid intimate-
media thickness was associated to increased TF and PAR2 expression
and markers of oxidative stress. Thrombosis development was further
associated with NO, peroxides, MMP, and GSH levels. Conclusions:
Oxidative stress could be a pathogenic link between aPL, thrombosis
and atherosclerosis development in APS. Antioxidant treatment might
have efficacy in preventing the clinical manifestations.
Disclosure: Supported by JA0042/2007, JA0246/2009, P08CVI04234
and PS09/01809.
A019
Clearance of Platelet Microvesicles and b2-glycoprotein I
Dasgupta S1, Abdel-Monem H1, Le A1, Prakasam A1, Thiagarajan P1
1
Baylor College of Medicine and Michael E. DeBakey Veterans Affairs
Medical Center, Houston, TX
Background: b2-Glycoprotein I is an anionic phospholipid-binding
50-kDa plasma protein. Its physiological role remains uncertain and
in vitro studies suggest a possible role in the clearance of anionic phos-
pholipid-containing membranes. A majority of the antiphospholipid
antibodies, present in patients with antiphospholipid syndrome, are
directed against conformational epitopes in b2-glycoprotein I.
Methods and Results: We labeled the carbohydrate moieties of b2-gly-
coprotein I with BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-inda-
cene)-hydrazide without affecting its phospholipid binding and
measured its effect on the phagocytosis of platelet microvesicles by
THP-1 cells derived macrophages. BODIPY- b2-glycoprotein I
bound to phosphatidylserine-expressing platelet microvesicles in a
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 Abstracts
502
concentration-dependent manner. Binding was inhibited by 50 fold
molar excess of unlabeled b2-glycoprotein I and annexin A5 and the
phosphatidylserine-binding C1C2 fragment of lactadherin. b2-glyco-
protein I also promoted the phagocytosis of platelet microvesicles by
THP-1 derived macrophages in vitro at physiological plasma concen-
trations with a half maximal effect at 10 ug/ml. b2-glycoprotein
I-stimulated phagocytosis is inhibited by annexin V and the C1C2
fragment of lactadherin. Furthermore, immunoaffinity purified
b2-glycoprotein I-dependent antiphospholipid antibodies from
5 patients inhibited the phagocytosis in a concentration dependent
manner. Conclusion: These studies suggest b2-glycoprotein I binding
to phosphatidylserine-expressing procoagulant platelet microvesicles
promotes their clearance by macrophages and autoantibodies to
b2-glycoprotein I inhibit the clearance to induce a procoagulant state
associated with these antibodies.
Disclosure: Nothing to disclose.
A020
The fine specificity of Anti-b2glycoprotein I antibodies in systemic
autoimmune diseases is preferentially directed against Domain I
Andreoli L1, Nalli C1, Motta M2, Norman GL3, Shums Z3, Encabo S3,
Binder W3, Avcin T4
1
Spedali Civili, Brescia, Italy; 2Neonatology and NICU, Brescia, Italy;
3
INOVA Diagnostics, Inc., San Diego; 4University Children’s Hospital,
Ljubljana, Slovenia; 5Ist. Gaetano Pini, Milan, Italy
Background: Anti-b2glycoprotein I antibodies (a-b2GPI) are detectable
not only in the Antiphospholipid Syndrome (APS), but also in systemic
autoimmune diseases (SAD) and healthy children (cld). A-b2GPI
against Domain1 (D1) associate with thrombosis, whereas those to
Domain4/5 (D4/5) were described in non-thrombotic conditions.
We evaluate the fine specificity of a-b2GPI in different groups of
adults and cld.
Patients: A) 59 with APS; B) 18 with SAD, no APS; C) 57 one-year-
old healthy cld born to mothers with SAD; D) 33 cld with Atopic
Dermatitis (AD). No thrombotic events in groups C&D. Methods:
All subjects were IgG a-b2GPI positive with routine home-made
ELISA. Sera were studied for IgG ab2GPI D1, D4/5 using research
ELISAs containing recombinant b2GPI antigens (QUANTA LiteTM,
INOVA Diagnostics). Results are in optical density (OD) values.
Results: Median OD is shown in the table.
a-2GPI IgG D1a-2GPI D4/5a-2GPI
(whole molecule) IgG IgG
A)APS 1.156 0.404 0.144
B)SAD,no APS 1.256 0.517 0.165
C)1-year-old cld 0.655 0.140 0.311
D)AD cld 0.726 0.148 0.224
Conclusion: Anti-b2GPI IgG in subjects with no systemic autoimmu-
nity nor APS (groups C&D) mostly recognize D4/5. This could
account for their innocent profile. A-b2GPI IgG with prevalent speci-
ficity for D1 seem to cluster in autoimmune conditions, including APS.
In patients without APS the pathogenic potential of IgG a-b2GPI D1
might have been mitigated by adequate prophylaxis or absence of
additional risk factors.
Disclosure: Norman, Shums, Encabo and Binder are employees of
INOVA Diagnostics.
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A021
Naturally occurring free thiols within b2-glycoproein i in vivo: functional
implications in the regulation of oxidative stress induced endothelial cell
injury
Ioannou Y1,2, Zhang JY1,3, Passam FH1, Rahgozar S1, Qi J1,
Giannakopoulos B1, Qi M1, Yu P1,3, Yu D3, Hogg P1, Krilis S1
1
UNSW, Sydney, Australia; 2UCL, London, United Kingdom;
3
TMU, Tianjin, China
Purpose: Biochemical study of the cysteine residues within b2-glyco-
protein I (b2GPI) reveals that some harbour an allosteric configuration
suggesting that under specific conditions, free cysteine thiols may exist.
We set out to investigate whether b2GPI ex vivo harbours free thiols
and aimed to assess the functional implications of this. Methods: A
novel capture ELISA employing a thiol specific reagent was developed
to detect b2GPI with free thiols within serum or plasma. The effect of
reduced versus non-reduced b2GPI on H202 induced cell death was
then quantified on both EAhy926 cells of endothelial cell lineage and
primary HUVEC. Results: Using this novel ELISA, it is shown for the
first time that b2GPI exists in vivo in a reduced state, upon screening 18
healthy human serum samples. Multiple negative controls, including
the use of b2GPI -/- serum confirmed excellent specificity for this
ELISA. Both the intra-plate and inter-plate coefficient of variation
was less than 10%. Incubating both EAhy926 cells and HUVEC
cells with H202 in the presence of reduced b2GPI resulted in a signifi-
cant abrogation of H202 induced cell death as compared to cells incu-
bated with H202 and non-reduced b2GPI or buffer alone. Conclusion:
This novel finding, that b2GPI exists in vivo in a reduced state, repre-
sents a paradigm shift as hitherto studies investigating the biology of
this autoantigen have employed purified protein which spontaneously
oxidises when fractionated from blood. With respect to endothelial
cells, reduction of b2GPI protects against oxidative stress induced
cell injury.
Disclosure: Nothing to disclose.
A022
Antibodies against domain I of b2-glycoprotein I are not associated with
arterial thrombosis in the general population
De Angelis V1, Siegerink B2, Algra A1, Rosendaal FR2, van Os G1,
de Groot PG1, Derksen RHWM1, Meroni PL3, Doggen CJM1,
Urbanus RT1
1
UMC Utrecht, Utrecht, The Netherlands; 2UMC Leiden, Leiden,
The Netherlands; 3University of Milan, Milan, Italy
Background: Clinically relevant antiphospholipid antibodies (aPL) are
mainly directed against b2-glycoprotein I (b2GPI). Several studies sug-
gest pathological aPL are directed against domain I (DI) of b2GPI.
Anti-DI antibodies were recently associated with an increased risk of
thrombosis in patients with aPL, but the contribution of anti-DI anti-
bodies to thrombotic risk in the general population remains unclear.
Aim: To determine the contribution of antibodies directed against DI
of b2GPI to the risk of arterial thrombosis in the general population.
Methods: We determined the risk of arterial thrombosis in relation
with anti-DI antibodies in two large population-based case-control
studies: The RATIO study, among 175 women with ischemic stroke
(IS), 203 with myocardial infarction (MI) and 628 healthy controls
who were all below age 50, and the SMILE study, which included
560 men with MI before the age of 70 and a control group of 646
men. We then measured anti-domain I antibodies in a cohort of anti-
phospholipid syndrome (APS) patients, including 39 patients with

primary APS and 9 with APS secondary to autoimmune disease. Fifty-
five autoimmune disease patients were controls. Results: In the RATIO
study, we found odds ratios (OR)(95%-confidence interval) of 1.26
(0.69-2.31) for MI and 0.72 (0.34-1.53) for IS. In the SMILE study
we found an OR of 0.95 (0.64-1.41) for MI. We then validated our
assay in APS patients and found that seven patients with APS (11%)
had anti-DI IgG, six (86 %) of whom had experienced thrombotic
events. One (1.8%) patient in the control group had anti-DI antibo-
dies. Conclusions: Anti-DI antibodies seem specific for patients with
the antiphospholipid syndrome and are not associated with an
increased risk of arterial thrombosis in the general population.
Disclosure: Nothing to disclose.
A023
Human endothelial cells secrete oxidoreductases on the cell surface and
can both nitrosylate and modulate the redox state of b2-glycoprotein I
Ioannou Y1,2, Zhang JY1,3, Passam F1, Rahgozar S1, Qi J1,
Giannakopoulos B1, Qi M1, Yu P1,3, Yu D3, Hogg P1, Krilis S1
1
UNSW, Sydney, NSW, Australia; 2UCL, London, United Kingdom;
3
TMU, Tianjin, China
Purpose: To investigate whether human endothelial cells may affect the
redox state of b2GPI and determine whether free cysteine thiols within
reduced b2GPI may be nitrosylated. Methods: The EAhy926 cell line of
human endothelial lineage and HUVEC were incubated with either
reduced b2GPI or non-reduced b2GPI (1 mM). The supernatant was
then labelled with a thiol specific reagent, transferred to a membrane
and blotted for thiol-labelled b2GPI. Western blot was also employed
to detect the presence of numerous oxidoreductase proteins in the
supernatant secreted by endothelial cells. Cysteine thiol nitroyslation
within b2GPI was confirmed by the established ‘biotin’ switch method
and also by using a specific anti-nitrosothiol antibody. Results: Both
EAhy926 cells and HUVEC were shown to significantly enhance free
thiol generation within b2GPI. Evidence for constitutive secretion by
HUVEC of PDI and other oxidoreductases, previously thought to only
reside in the ER, were demonstrated - notably TRX-R, ERp46 and
ERp72. b2GPI, once reduced, could then undergo cysteine nitrosyla-
tion directly by LPS stimulated HUVEC and also by direct incubation
with S-nitrosoglutathione. Conclusions: These findings support the
concept of a dynamic complex interplay between cell membrane
bound proteins and oxidoreductases on the surface of endothelial
cells. Given that reduced b2GPI serves to protect endothelial cells
against oxidative stress, the ability of these cells to enhance the reduc-
tion of b2GPI and nitroyslate free cysteines suggests a regulatory role
for b2GPI in modulating the oxidative micro-environment at the cell
surface.
Disclosure: Nothing to disclose.
A024
Beta2-glycoprotein I directly modulates the level of plasma cholesterol
in vivo
Giannakopoulos B1, Qi JC1, Miao Q1, Matthaei K2, Rye KA3, Krilis S1
1
STGPH, UNSW, Sydney, NSW, Australia; 2JCSMR, ANU,
Canberra, ACT, Australia; 3HRI, Sydney, NSW, Australia
Aim: The aim of this study is to delineate the function of b2GPI with
regards to lipid metabolism using b2GPI knockout (KO) mice.
Method: Our lab has generated b2GPI KO mice on a C57BL6/J back-
ground. At 4 months of age, 10 of each sex; b2GPI KO males and
Abstracts
503
females, and control males and females were commenced on a high fat
diet. At 8 months the mice were weighed and then sacrificed. Total and
HDL cholesterol and triglyceride (TG) levels were determined. Non-
HDL cholesterol was calculated by subtracting HDL from total cho-
lesterol levels. Results: a) b2GPI deficient females weigh significantly
more than controls: P value ¼0.01 b) b2GPI deficient female livers
weigh significantly more than those of controls: P value ¼0.003
c) b2GPI deficient females have significantly higher total cholesterol
levels compared to controls: 3782 þ/-327.6 vs 2547 þ/- 86.7 mM, P
value ¼0.006 d) b2GPI deficient females have significantly higher HDL
cholesterol levels compared to controls: P value ¼0.0367 e) b2GPI
deficient females have significantly higher non-HDL cholesterol levels
compared to controls: P value¼0.0032 f) b2GPI deficient females do
not have statistically significant different TG levels compared to con-
trols: P value ¼0.891 g) b2GPI does not modulate weight, cholesterol,
and TG levels in the males of this strain. Conclusion: In female mice
(C57BL6/J strain) fed a high fat diet, b2GPI deficiency leads to higher
levels of total, HDL and non HDL cholesterol, accompanied by
greater weight gain, and larger livers. These results, in conjunction
with previous in vitro studies that demonstrate that b2GPI can directly
bind to receptors of the LDL receptor family including megalin, and
previous findings that b2GPI is contained in plasma lipid fractions,
suggest that b2GPI partakes a physiological role in lowering choles-
terol levels in the plasma, perhaps by enhancing uptake of LDL and
HDL cholesterol via b2GPI- LDL receptor interactions. Whether this
mechanism is disrupted in APS patients predisposing to accelerated
atheroma formation requires further investigation.
Disclosure: Nothing to disclose.
A025
Beta 2 glycoprotein I is a substrate of thiol oxidoreductases
Passam F1, Rahgozar S1, Giannakopoulos B1, Qi M1, Tanaka K1,
Hogg P2, Wong J2, Raftery M2, Krilis S1
1
University of New South Wales, St. George Hospital, Sydney, NSW,
Australia; 2University of New South Wales, Wallace Wurth Building,
Sydney, NSW, Australia
Purpose: Analysis of the crystal structure of b2GPI predicted disulfide
bonds with the potential to participate in reactions with thiol oxidor-
eductases. In this study we determined cysteines in the b2GPI molecule
that can be involved in thiol exchange reactions. Methods: b2GPI was
reduced with thioredoxin-1 (TRX-1) and protein disulfide isomerase.
Free thiols formed in b2GPI were labeled with Na-(3-maleimidylpro-
pionyl) biocytin (MPB), a biotin thiol specific probe. The labeled
cysteine residue(s) in the b2GPI molecule were detected by mass spec-
trometry. Results: b2GPI contains 11 disulfide bonds and no unpaired
cysteine. The fifth domain has an extra 20 amino acid tail with an
unusual termination in a cysteine, that forms a loop-back disulfide
link with an extra cysteine found midway between the standard
second and third cysteine residue positions. The introduction of free
thiols into b2GPI was shown by the labeling with MPB after reaction
with TRX-1 and PDI. Biotin labeling of reduced beta 2 glycoprotein I
identified Cys326 in domain V as the predominant cysteine involved in
the thiol exchange reaction with TRX-1. Conclusion: This study is the
first to show that b2GPI can be involved in thiol exchange reactions.
The identification of a reactive cysteine in the fifth domain is in agree-
ment with the majority of biological functions of b2GPI being attrib-
uted to this domain (i.e phospholipid, thrombin and gpIba binding).
Disclosure: Nothing to disclose.
Lupus
 Abstracts
504
A026
Reduction of beta 2 glycoprotein I by thioredoxin-1 promotes binding
to von Willebrand factor
Giannakopoulos B1, Rahgozar S1, Passam F1, Gemmell R1, Qi M1,
Qi JC1, Krilis S1
1
St. George Hospital, Sydney, NSW, Australia
Purpose: Our group has shown that beta 2 glycoprotein I (b2GPI) can
be reduced by thioredoxin-1-and protein disulfide isomerase. The pur-
pose of this current study was to determine if the reduced form of
b2GPI could affect its binding to vWF. Methods: Binding of reduced
b2GPI by TRX-1 to immobilized vWF was performed on 96 well
microtiter plates coated with human vWF. vWF activated with risto-
cetin was added to immobilized b2GPI. Recombinant gpIba was
added to vWF coated wells in the presence of TRX-1 reduced
b2GPI. Results: Reduced b2GPI by TRX-1 showed significantly
higher binding to vWF than non reduced b2GPI. The increased
binding of reduced b2GPI to vWF was inhibited when free thiols intro-
duced into b2GPI by TRX-1 had been blocked with Na-(3-maleimidyl-
propionyl) biocytin, a biotin thiol specific probe. The amount of b2GPI
bound to vWF was significantly reduced when the TRX-1 reductase
activity had been inactivated with 1-chloro-2,4-dinitrobenzene.
Ristocetin-activated vWF compared to non-activated vWF had a
marked increase in binding to reduced b2GPI. Binding of recombinant
gpIba to immobilized vWF was increased in the presence of reduced
b2GPI compared to non-reduced b2GPI. Conclusion: Given the evi-
dence of the pivotal role of thiol oxidoreductases, such as protein
disulfide isomerase, in platelet thrombus formation we propose a
potential regulatory role where beta 2 glycoprotein I in its reduced
state may promote gpIba binding to vWF.
Disclosure: Nothing to disclose.
A027
Exploring the interaction between anti b2-Glycoprotein I (b2GPI)
antibodies and Domain I of b2GPI in high risk patients with APS
Banzato A1, Pozzi N1, De Filippis V1, Bison E1, Bontadi A1, Hoxha A1,
Ruffatti A1, Pengo V1
1
University of Padova, Padova, Italy
Background: Antiphospholipid syndrome (APS) comprises an hetero-
geneous group of patients with thrombosis or pregnancy morbidity
and different antiphospholipid (aPL) antibody profiles (Pengo V,
T&H 2005). As the possible pathogenic antibody in APS is that direc-
ted toward the Domain I of b2-Glycoprotein I (b2GPI), this study is
aimed to explore the interaction of anti b2GPI antibodies with syn-
thetic Domain I (DmI) in high risk patients. Methods: We chemically
synthesized the full-length correctly-folded b2GPI/DmI (1-64) and
linear DmI peptides (Gly40-Leu52 and Thr57-Ile70) to set up direct
binding and competitive inhibition enzyme-linked immunosorbent
assays (ELISA). The interaction was then tested by using dot blot
analysis. Plasma of 23 patients with APS and triple positivity
(LACþ, IgG aCLþ, IgG ab2GPIþ) were evaluated. Results: Direct
binding experiments with b2GPI/DmI coated into ELISA plates did
not prove any interaction with anti-b2GPI autoantibodies. The same
was found using linear peptides. Competition ELISA with b2GPI-
coated plates and b2GPI/DmI in solution phase, demonstrated a
dose-dependent inhibition of anti b2GPI autoantibodies binding in
patients with triple positivity at the b2GPI:b2GPI/DmI molar ratio
of 1:0.1, 1:0.5, 1:1, 1:5, respectively. Percentage inhibition at a molar
ratio 1:5 varied from 10% to 50%. No inhibition was observed by
using the nonspecific protein antithrombin or DmI linear peptides.
Anti-b2GPI autoantibodies binding to DmI was confirmed in dot
Lupus
blot experiments where optical density in chemiluminescence between
triple positive APS patients and normal controls was significantly dif-
ferent (ratio patients/controls¼ 4, p¼0.02). Conclusions: Anti-b2GPI
autoantibodies bind to correctly folded synthetic DmI in solution and
in a dot blot system but not to DmI adsorbed into ELISA plates. DmI
linear peptides do not retain this inhibition capacity.
Disclosure: Nothing to Disclose.
A028
Low levels of IgM antibodies to oxidized cardiolipin but not to
cardiolipin increase but high levels decrease risk of cardiovascular
disease: pro-inflammatory effects of oxidized cardiolipin
Hua X1, Su J1, deFaire U1, Frostegård J1
1
Karolinska Institutet, Stockholm, Sweden
Objective: Cardiolipin CL is present in mitochondria and bacteria, as
well as in lipoproteins and can easily oxidized, with two double bonds.
Here we study potential pro-inflammatory properties of oxidized CL
(oxCL) and inhibitory effects of Annexin A5, an antithrombotic agent
interfering with phospholipids. Further, little is known about the prop-
erties of oxCL and a putative role of antibodies against CL (aOxCL).
Method: Sixty-year olds from the Stockholm County were included
in a screening (2039 men and 2193 women), 211 incident cases of
CVD (myocardial infarction, ischemic stroke, or hospitalized angina
pectoris) and 633 age- and sex-matched controls were identified
through a 5-7 year follow-up. Serum levels of antibodies was deter-
mined by ELISA (in house method for aOxCL and commercial kit
for aCL. Effects of oxCL were determined by FACS and ELISA.
Result: oxCL but not CL induced activation of EC, effects inhibited
by Annexin A5. aOxCL levels below 50th to 10th percentile, adjusted
for smoking, BMI, type II diabetes, hypercholesterolaemia, and high
blood pressure were associated with increased risk of CVD up to three
times (p<0.01). Maximal risk was determined with cut-off at 30th
percentile. On the contrary, high levels were associated with decreased
risk of CVD, reaching significance over 82% with a RR at 0.57
(p¼0.01). In subgroup analysis, MI in both men and women showed
similar results, with increased risks at low levels. For men, there was a
striking association with low levels of anti-oxCL increasing the risk
independent of other risk factors 12 fold (p<0.01). In contrast, aCL
did not function as a protection factor at high levels or risk factor at
low levels. Conclusions: OxCL has novel inflammatory properties.
aOxCL could be a novel risk marker novel risk marker for CVD.
Active or passive immunization to raise antibody levels are of interest
as novel therapeutic concepts.
Disclosure: UdF and JF are named as inventors on patents relating to
oxidized phospholipids.
A029
The role of oxidized-LDL/B2-glycoprotein I complexes and anti-B2GPI
antibodies in autoimmune-mediated atherosclerosis
López LR1, Greco TP2, Conti-Kelly AM2, Anthony JR2, Boisen M1,
Kojima K3, Matsuura E4
1
Corgenix, Inc., Broomfield, CO; 2Saint Mary’s Hospital, Waterbury, CT;
3
Medical and Biological Laboratories Co, Ltd., Ina, Nagano, Japan;
4
Okayama University Graduate School of Medicine, Okayama, Japan
Oxidized-LDL/b2-glycoprotein I complexes (oxLDL/b2GPI) and anti-
phospholipid antibodies (aPL) have been implicated in atherogenesis.
aPL predict cardiovascular adverse events (AE) and high titers of
oxLDL/b2GPI are associated with severe disease and EA in patients
with acute coronary syndrome (ACS). OxLDL/b2GPI and aPL

(anti-b2GPI and anti-oxLDL/b2GPI) were measured to evaluate their
role on the severity of coronary artery disease (CAD) and AE in 339
ACS patients. Patients with severe CAD (graded angiography) had
significantly higher mean titers of oxLDL/b2GPI and more AE than
patients with normal coronaries. The occurrence of AE increased sig-
nificantly in aPLþ patients with severe disease (grades IV-VI) than
aPL- patients (30.6% vs. 9.0%, p<0.005). The relative risk (RR) for
AE of all aPLþ versus aPL- patients was 3.66 (95% CI 1.88-7.14,
p<0.001). aPLþ patients with oxLDL/b2GPI in the 2 upper quartiles
had significantly more AE than aPL- patients in the same quartiles
(26.1% vs. 6.0%, p<0.05) with a RR of 3.2 (95% CI 0.95-10.86,
p<0.05). RR in aPLþ patients in the 2 upper quartiles was 6.72
(95% CI 2.4-19.0, p¼0.01) when compared to aPL- patients in the 2
lower quartiles. The RR of aPLþ patients in the upper oxLDL/b2GPI
quartile increased to 13.9 (95% CI 1.8-102.4, p<0.001) when compared
to aPL- patients in the lowest quartile. The co-presence of oxLDL/
b2GPI complexes and aPL (anti-b2GPI and anti-oxLDL/b2GPI) is
associated with CAD severity and significantly enhances the risk of
cardiovascular AE. These findings support the concept that autoimmu-
nity accelerates atherosclerosis.
Disclosure: L Lopez and M Boisen are employees of Corgenix, Inc. K
Kojima is employee of Medical and Biological Laboratories Co, Ltd.
A030
Structural and binding studies of b2GPI with lipoprotein receptors and
cardiolipin
Lee CJ1, De Biasio A1, Beglova N1
1
Harvard Medical School, Boston, MA
Background: b2GPI is the major antigen for APS-related antibodies. It
is known that b2GPI binds all receptors of the LDLR family and the
binding site for receptors is located in domain V of b2GPI (b2GPI-
DV). ApoER2, a member of the LDLR family, is involved in platelet
activation by b2GPI/antibody complexes. Other lipoprotein receptors
besides ApoER2 are expressed on endothelial cells and monocytes and
might contribute to the activation of these cells by b2GPI/anti-b2GPI
complexes in APS. Questions to answer: The ligand-binding domains of
lipoprotein receptors are formed by multiple LA modules. Where is the
binding site for LA modules on b2GPI? Is the binding site the same for
different LA modules? How can LA modules that bind b2GPI be dis-
tinguished? Methods and Results: We measured and compared the
binding affinity of different LA modules for b2GPI-DV and mapped
the binding site for LA modules on b2GPI-DV by solution NMR
spectroscopy. Using molecular docking guided by NMR-derived
restraints, we built a model of a complex between b2GPI-DV and an
LA module. We also measured the binding of b2GPI-DV to cardiolipin
and observed that LA modules interfere with this binding.
Disclosure: This presentation was made possible by a support from
the American Society of Hematology and the American Heart
Association.
A031
Binding Characteristics of Anti-b2GPI Paratopes Revealed by Phage
Display
Zager U1, Lunder M2, Cucnik S1, Strukelj B2, Kveder T1, Rozman B1,
Bozic B1,2
1 staging AD.
University Medical Centre, Ljubljana, Slovenia; 2University of
Ljubljana, Ljubljana, Slovenia
Abstracts
505
Objectives: It was reported that hydrogen bonds are important for
antib2GPI-b2GPI interaction. Furthermore, sequence analysis of
antib2GPI has shown the presence of positively charged residues
(R, K) in their CDRs, suggesting an important role for charge inter-
actions in the binding of antib2GPI to antigen.
The aim of our study was to determine the binding characteristics
of antib2-GPI paratopes by phage display method. Materials and
Methods: A linear heptamer phage display library (NewEngland
BioLabs) was screened with b2GPI. Monoclonal anti-b2GPI, purified
high and low avidity antib2GPI IgG derived from plasma of an APS
patient and oxidized native IgGs from blood donors were used as
specific eluents. Single-stranded DNA from amplified selected phage
clones was isolated and sequenced (MWG Biotech, Germany). Results:
Selection of b2GPI binding peptide sequences by phage display yielded
a series of similar sequences regardless of the antib2GPI IgG subgroup
used for elution. The common motif was established. Binding affinity
of selected peptides was confirmed with ELISA. Conclusions: Motifs
with high affinity for b2GPI were revealed by screening of the linear
heptamer peptide phage display library using of antib2GPI IgG as
eluents. They represent putative amino acids in paratopes of mono-
clonal antib2GPI, purified high and low avidity antib2GPI IgG and
oxidized native IgGs, and demonstrate importance of electrostatic
bonds for their interactions with b2GPI.
Disclosure: Nothing to disclose.
A032
Serum redox-reactive antiphospholipid antibodies may serve as a blood
biomarker for Alzheimer’s disease
Wagenknecht DR1, Ramsey C2, McIntyre JA1,3
1
St. Francis Hospital, Beech Grove, IN; 2Alternate Hypothesis,
Indianapolis, IN; 3Indiana University-Purdue University Indianapolis,
Indianapolis, IN
Increased oxidative stress in the central nervous system (CNS) of
Alzheimer’s disease (AD) patients results in oxidation of proteins,
lipids and DNA. Oxidative changes may be a consequence of an abnor-
mal enrichment of redox-reactive metals in AD brains. Redox-reactive
antibodies (R-RAA) are a family of autoantibodies detectable only
after exposure to oxidizing agents. We reported significantly decreased
or absent R-RAA in the cerebrospinal fluids of autopsy-confirmed AD
patients (Autoimmunity 2007; 40:390). We next queried if R-RAA in
the blood of AD patients differ from R-RAA levels in age-matched
controls. We compared 16 AD patients to 17 volunteer blood bank
donors. Each specimen was tested before and after hemin-induced
oxidation by using an in-house ELISA to detect IgG, IgM and IgA
to 4 aPL specificities; aCL, aPS aPE and aPC. Significant aPE differ-
ences were found between the R-RAA in blood from AD patients
compared to normal donors. In-sample Fisher’s linear discriminate
analysis found 88% sensitivity and 94% specificity for predicting AD
from serum. In-sample Classification and Regression Tree analysis
(CART) found a sensitivity of 84% and a specificity of 100% for
predicting AD. Decreased aPE R-RAA in AD serum is consistent
with decreased circulating levels of ethanolamine plasmalogen in AD
patients (Goodenowe, JLR; 2007; 48:2485). We are currently testing 18
blinded NIH-ADNI serum samples from AD, mild cognitive impair-
ment (MCI) and normal individuals to determine if an R-RAA blood
test can be used as a laboratory biomarker for diagnosing and/or
Disclosure: Nothing to disclose.
Lupus
 Abstracts
506
A033
Downregulation of KLF2 by Antiphospholipid Antibodies Affects NF-iB
Signaling and is Modulated in Part by CBP
Allen K1, Kawanami D1, Jain M1, McCrae K1
1
Case Western Reserve University, Cleveland, OH
Antiphospholipid syndrome (APS) is characterized by recurrent
thrombosis and pregnancy loss in the presence of antiphospholipid
antibodies (APL). b2-glycoprotein I (b2GPI) is a primary target of
APL. Anti-b2GPI antibodies activate endothelial cells, enhancing the
expression of adhesion molecules and tissue factor, and the secretion of
proinflammatory cytokines. Kruppel-like factors (KLF) regulate
endothelial cell inflammatory responses. KLF2 mediates anti-athero-
sclerotic and anti-inflammatory effects in endothelial cells; it is induced
by laminar flow, inhibited by inflammatory cytokines, and its over-
expression blocks endothelial cell expression of VCAM-1 and E-selec-
tin caused by TNF. We showed in vitro that endothelial cell activation
induced by APL/anti-b2GPI antibodies results in downregulation of
KLF2 and, as also reported by others, activation of NF-kB.
Overexpression of KLF2 in endothelial cells partially blocks cell acti-
vation by APL as measured by E-selectin ELISA and confirmed by
E-selectin luciferase reporter assay. Western blot analysis shows that
overexpression of KLF2 does not prevent the phosphorylation of
NF-kB p65 subunit serine 536 in endothelial cells activated by APL.
However, KLF2 overexpression studies in APL-activated endothelial
cells show that KLF2 inhibits NF-kB transcriptional activity as mea-
sured by luciferase reporter assay. The inhibition of NF-kB signaling
by KLF2 appears to be due to recruitment of the CBP cofactor away
from NF-kB by KLF2 as preliminary data shows that restoring the
pool of available CBP by overexpression tilts the KLF2/NF-kB bal-
ance back toward NF-kB. These data suggest that APL influence
KLF2 expression in vitro and and that modulation of KLF2 by APL
may contribute to the prothrombotic and proinflammatory phenotype
seen in APS.
Disclosure: Nothing to disclose.
A034
Variable effects of an Inhibitor of Nuclear Factor-Kappa B on
antiphospholipid (aPL) antibody-mediated effects in vivo
Montiel-Manzano G1, Romay-Penabad Z1, Papalardo E1,
Aguilar-Valenzuela R1, Pierangeli SS1
1 phages when compared to IgG-APS þ CP group(p¼ 0.034,0.036,
UTMB, Galveston, TX
Background: aPL are a heterogeneous group of antibodies. It has been
shown that endothelial cell (EC) activation and tissue factor (TF) up
regulation in EC and monocytes by aPL leads to a pro-thrombotic
state and involves translocation of NF-kB. Here, we studied the effects
of a NF-kB inhibitor on the thrombogenic effects of aPL antibodies
in vivo. Methods: C57BL/6J mice were injected with IgG (IgG-APS,
n¼5) or IgM-APS, n¼2) isolated from seven patients with APS or
from healthy control subject (IgG-NHS or IgM-NHS). Two groups
of mice were injected intraperitoneally (i.p.) with either 10 mM of
MG132 (an NF-kB inhibitor), or with 60/40 DMSO/PBS (media) 30
min before the Ig’s injections. Each of these two initial groups was
equally divided in two further subsets to receive i.p. either 500 mg of
IgG-APS or IgM-APS or IgG-NHS or IgM-NHS twice (at time 0 and
48 h later). Seventy-two hours after the first injection, the following
procedures were done in each mice: a) dynamics of thrombus forma-
tion (thrombus size), b) TF function in homogenates of carotid arteries
and peritoneal cells, c) TF expression in the aortas using quantum dot
nanocrystal and two-photon excitation laser scanning microscopy, d)
soluble E-selectin by ELISA. Results: All IgG-APS and IgM–APS
Lupus
increased significantly thrombus size, TF activity (carotid arteries
and peritoneal macrophages and TF expression in aorta of mice
when compared to control Ig-treated mice. sE-selectin was increased
in the serum of all mice treated with Ig-APS when compared to con-
trols. Importantly, MG132 significantly inhibited thrombus size, TF
expression and TF activity in homogenates of aortas and in peritoneal
macrophages and sE-selectin in mice injected with four out of the seven
Ig-APS. Conclusions: Some but not all thrombogenic effects of aPL
antibodies in vivo are mediated by NF-kB. These studies underscore
the heterogeneous nature of aPL antibodies and their pathogenic
effects. Utilizing NF-kB inhibitors may be a novel strategy to amelio-
rate the pathogenic effects of some subgroups of aPL antibodies.
Disclosure: Nothing to disclose.
A035
Abrogation of aPL pathogenic effects by a C5a receptor (C5aR)
antagonist
Aguilar-Valenzuela R1, Carrera-Marin AL3, Romay-Penabad Z1,
Papalardo E1, Garcia E3, Lambris J2, Qu HC2, Pierangeli SS1
1
University of Texas Medical Branch, Galveston, TX; 2University Of
Pennsylvania, Philadelphia, PA; 3Instituto Polite´cnico Nacional,
Ciudad de Mexico, Distrito Federal, Mexico
Background: The complement system has been shown to be involved in
antiphospholipid antibodies (aPL) pathogenicity, obstetrics and
thrombotics. Complement split products (C5a and C3a) may be pro-
duced when aPL antibodies bind to target cells. Those in turn may
enhance endothelial cell activation and a pro-infllammatory/procoagu-
lant state. Objective: To examine the effects of a C5a receptor antago-
nist peptide (C5aR-AP) on aPL-mediated thrombosis, in vivo.
Methods: C57BL/6J mice were injected i.p. twice with 500 mg of IgG
from a patient with Antiphospholipid syndrome (APS) (IgG-APS) or
with control IgG (IgG-NHS) preceded by i.p. injection of 25 mg of
C5aR-AP (Ac-Phe-c[Orn-Pro-DCha-Trp-Arg] or control peptide
(CP) (Ac-Phe-c[ORn-Pro-DCha-Ala-DARg]). Seventy-two hours
after the first injection, the size of an induced thrombus was measured
as well as tissue factor activity was determined in carotid arteries
homogenates and in peritoneal macrophages. Results: IgG-APS þ
CP induced significantly larger thrombi, TF activity in carotids and
macrophages compared to IgG-NHS þ CP treated mice (p¼0.027,
0.045, 0.063). The IgG-APS and C5aR-AP treated group had signifi-
cantly smaller thrombi and lower TF activity in carotid and macro-
0.045). Conclusions: The data support the involvement of complement
system in APS thrombotic complications and underscore the possibi-
lity of utilizing complement inhibitors as alternative to ameliorate
them.
Disclosure: Nothing to disclose.
A036
Anti-b2 glycoprotein I antibodies from leprosy patients do not show
thrombogenic effects in an in vivo animal model
Forastiero R1, Martinuzzo M2, de Larrañaga G3, Vega-Ostertag M4,
Pierangeli SS4
1
Favaloro University, Buenos Aires, Argentina; 2Favaloro Foundation,
Buenos Aires, Argentina; 3Hospital Muñiz, Buenos Aires, Argentina;
4
The University of Texas Medical Branch, Galveston, TX
Background: The APS-associated aPL are autoantibodies directed
against b2GPI and prothrombin. Patients with leprosy present high
frequency of IgM aPL that bind b2GPI, but they do not develop

thrombosis. Anti-b2GPI from APS mainly bind to domain I of b2GPI
while in leprosy are directed against domain V. There is convincing
evidence that aPL/anti-b2GPI from APS are pathogenic in vivo and in
vitro. Objective: To investigate the thrombogenic and pro-inflamma-
tory effects of aPL from leprosy and to compare with aPL from APS.
Methods: Sera from 6 patients with APS and 6 with leprosy were used
as the source of IgM. All APS and 5 leprosy patients had strong LA
activity, and high titers of IgM aCL/anti-b2GPI. The remaining
leprosy patient was aPL negative (control). We treated CD1 mice, in
groups of 5, at 0 hours and 48 hours later with IgM aPL/anti-b2GPI
isolated from patients with leprosy (IgM-leprosy) or APS (IgM-APS)
or with IgM from 2 healthy controls (IgM-NHS) or 1 leprosy aPL
negative control. Seventy-two hours after the first injection, the adhe-
sion of leukocytes (#WBC) to endothelial cells (EC) in cremaster
muscle (as an indication of EC activation in vivo), as well as the size
of an induced thrombus in the femoral vein of the mice were examined.
Results: IgM-APS significantly increased the thrombus size (in mum2)
when compared to IgM-NHS or IgM-leprosy treated mice (p<0.001).
There was no difference in mice injected with IgM-leprosy, IgM-NHS
or IgM-leprosy control. IgM-APS increased the #WBC adhering to
EC, when compared to IgM-NHS or IgM-leprosy (p<0.001).
IgM-leprosy
IgM-APS IgM-leprosy IgM-NHS control
Mean thrombus size 4446 1378 748 851
#WBC 5.1  2.2 2.0  1.0 3.5  1.6 1.3  0.5
Conclusions: Our data shown that aPL/anti-b2GPI from leprosy
patients have not thrombogenic and pro-inflammatory effects in vivo
when compared with aPL derived from APS.
Disclosure: Nothing to disclose.
A037
Autoantibodies against the Fibrinolytic Receptor, Annexin A2, in
Cerebral Venous Thrombosis
Cesarman Maus G1, Cantú C2, Barinagarrementerı´a F3, Villa R2,
Reyes E4, Hajjar K5, Garcı´a-Latorre E4
1
INCan, D.F., Mexico; 2INCMNSZ, D.F., Mexico;
3
Angeles, Queretaro, Mexico; 4Polite´cnico Nacional, D.F, Mexico;
5 Disclosure: Nothing to disclose.
Weill Medical College, New.York, NY
Background: Antibodies against annexin A2 (a pro-fibrinolytic
endothelial cell surface receptor that binds plasminogen, its tissue acti-
vator, and is the high affinity receptor for b2-GPI), are associated with
antiphospholipid syndrome (APS), and are able to activate monocytes,
umbilical vein endothelial cells, and inhibit endothelial cell surface-
localized plasmin generation independently of b2-GPI. Here we evalu-
ate the prevalence of anti-A2 antibodies in a cohort of individuals with
CVT. Methods: Serum samples from 185 individuals, including 40(33
women/ 7 men) with CVT and 145 healthy subjects, were analyzed by
ELISA and by immunoblot for classical antiphospholipid antibodies
(aPLA) (anticardiolipin, lupus anticoagulant, anti-b2-GPI) and anti-
A2. Patients were also studied for hereditary and acquired prothrom-
botic factors. Results: One or more well characterized prothrombotic
risk factors were present in 85% of individuals with CVT (pregnancy/
post-partum 57.5%, aPLA 22.5%, hereditary risk factors 20%). Anti-
A2 antibodies (titer >3SD) were significantly more prevalent in
patients with CVT (12.5%: IgG 7.5% and IgM 5%), than in healthy
individuals (2.1% of subjects: IgG 1.4% and IgM 0.7%; p<0.01).
Forty percent of individuals with anti-A2 had APS, while anti-A2
antibodies were present as the sole prothrombotic risk factor in 7%
Abstracts
507
of CVT patients. Conclusions: This study shows that antibodies against
the fibrinolytic receptor annexin A2 are significantly associated with
CVT, may be detected in the absence of classical aPLA, and may be a
marker of underlying immune-mediated thrombosis.
Disclosure: Nothing to Disclose.
A038
Involvement of C6 and the MAC on in vivo antiphospholipid-mediated
pathogenic effects
Carrera Marin AL1, Aguilar-Valenzuela R1, Romay-Penabad Z1,
Papalardo E1, Martinez-Martinez LA1, Pierangeli SS1
1
University of Texas Medical Branch, Galveston, TX
Background: Complement activation plays a role in pathogenesis of the
Antiphospholipid Syndrome (APS). However whether C5b-9 MAC is
also involved, is unknown. We addressed that question by studying the
effects of human aPL antibodies on thrombosis and tissue factor (TF)
upregulation in C6 deficient (-/-) mice. Methods: C6 deficient (-/-) mice
and the corresponding wild-type (WT) mice C3H/HeJ (C6þ/þ) were
injected with IgG isolated from a patient with APS (IgG-APS) or with
the corresponding control IgG-NHS. Then, the size of induced
thrombi in the femoral vein, IgG anticardiolipin (ACL) activity in
the sera were determined. TF activity was determined in homogenates
of carotid arteries and in peritoneal macrophages. Results: Thrombus
sizes were significantly larger in WT mice C6 (þ/þ) treated with IgG-
APS when compared with C6 (þ/þ) mice treated with IgG-NHS
(p¼ <0.001) The sizes of thrombi were significantly smaller in the C6
(-/-) mice injected with IgG-APS compared to their WT C6 (þ/þ)
counterparts (p¼ <0.001) showing an important abrogation of throm-
bus formation in mice lacking C6. Thrombus sizes in C6 (-/-) mice
injected with IgG-NHS were not different when compared to C6
(þ/þ) mice treated with IgG-NHS (p¼ 0.786). Similarly, upregulation
of TF activity in carotid arteries homogenates and in peritoneal macro-
phages in C6 (þ/þ) mice treated with IgG-APS were significantly
diminished in C6 (-/-) mice (p¼0.036 and p¼0.041, respectively). All
mice injected with IgG-APS had medium-high titers of aCL in their
serum. Conclusions: These data indicate that the C6 component of the
complement system mediates aPL-thrombogenic effects, underscoring
the importance of complement activation by aPL antibodies.
A039
Elevated Cytokines and Chemokine Levels in Antiphospholipid Antibody
(aPL)-Positive Patients
Martı´nez-Martı´nez LA1, Aguilar-Valenzuela R1, Doan E1,
Papalardo E1, González E1, Murthy V1, Seif A1, Erkan D2,
Pierangeli SS1
1
University of Texas Medical Branch, Galveston, TX; 2Hospital for
Special Surgery, New York, NY
Background: Certain cytokines and chemokines are believed to be
associated with antiphospholipid syndrome (APS). Purpose:
Compared to controls, to examine the levels of cytokines/chemokines
in aPL-positive patients. Methods: Interleukin (IL)-1b, IL6, IL8, TNF-
a, VEGF, (interferon inducible protein (IP)-10, sCD40L were mea-
sured in serum using a Multiplex Assay; sE-sel, and sTF were detected
by ELISA in the serum of aPL þ patients and in controls. Results:
Table demonstrates the biomarker levels of 22 aPL-positive patients
compared to controls. In a subgroup analysis, SLE patients with
Lupus
 Abstracts

508
aPL/APS (n¼10) had significantly higher levels of TNF-a, IP10 and complement inhibitory effects. AP inhibition occurred at 5-10 fold
sCD40L compared to aPL/APS patients without SLE (n¼12) (p¼ 0.05, therapeutic levels, while pathways using the classical components
0.029 and 0.05, respectively). may potentially be inhibited by adjustment of current clinical thera-
peutic target levels (in excess of 2 IU/ml).
Means of aPL þ Means of Disclosure: Nothing to disclose.
Biomarkers samples controls p value

IL-1b 7.65 0.35 <0.0001 A041

IL-6 42.15 0.71 <0.0001
Il-8 27.44 40.9 0.1763
TNF-a 14.71 0.46 <0.0001
VEGF 208.78 113.59 0.9778
IP-10 683.43 106.7 <0.0001
sCD40L 1070.19 24.67 <0.0001
sTF 449.59 13.05 <0.0001
sE-sel 20.78 42.04 0.0006
Conclusions: Our results underscore the importance of biomarkers in
aPL-positive patients that may help better understand pathogenic
mechanisms and develop targeted treatments.
Disclosure: Nothing to disclose.
A040
Inhibition of Complement Activation by Heparins But Not Fondaparinux
in the Alternative, Mannose Binding Lectin, and Classical Complement
Pathways
Ramirez AA1, Myones BL1
1
Baylor College of Medicine, Houston, TX
Background: Complement activation has been implicated in
Antiphospholipid Syndrome disease pathogenesis. LMWH have been
used to treat thrombosis and microangiopathic processes.
Unfractionated heparin (UFH) and LMWH have also been implicated
in classical pathway complement inhibition. Purpose: Evaluate effects
of UFH, LMWH and direct thrombin inhibitor, fondaparinux, on cell
membrane-associated classical (CP), mannose/mannan binding lectin
(MBL), and alternative pathway (AP) activation in vitro and in vivo.
Methods: AP-MBL-CP modified hemolytic assays. Unsensitized rabbit
erythrocytes (RaE) (direct activator human AP) in GVB/MgEGTA
(blocks CP activation). Sheep erythrocytes (ShE) sensitized with
mannan by CrCl3 in MBL assay (GVB/Caþþ/Mgþþ). Hemolysin
(anti-ShE IgM) sensitized ShE in CP hemolytic assay. Varying doses
of UFH, enoxaparin (0.5-10 IU/ml final conc) and fondaparinux (1.25-
6.25 mg/ml) with normal human serum as complement source.
Incubated 15-30 min @370C. Supernant OD541 expressed as % lysis
against controls. Results: Escalating doses of enoxaparin or UFH
resulted in increased % inhibition of complement-mediated RaE or
ShE lysis (1mg enoxaparin  100 IU anti-Factor-Xa activity) whereas
fondaparinux had no appreciable effect. AP: 0.5 IU/ml enoxaparin or
UFH < 1.5%, 2 IU/ml < 5%, 10 IU/ml < 55% inhibition of RaE lysis.
MBL - CP: 0.5 IU/ml UFH or enoxaparin < 30%, 2 IU/ml < 67%, 10
IU/ml < 95% inhibition of ShE lysis. UFH and LMWH curves super-
impose above 5 IU/ml. A small % inhibition of membrane-associated
complement activation was noted at anti-factor-Xa activity levels
achievable with standard therapeutic anticoagulant regimens of UFH
or LMWH, but LMWH > UHF achieved significant inhibition at
higher (>2 IU/ml) dosing levels for CP > AP components.
Preliminary data utilizing serum from patients on standard anticoagu-
lant doses showed minimal inhibition. Conclusions: UFH and LMWH
(enoxaparin) inhibit membrane-associated complement activation via
all 3 pathways in vitro while fondaparinux, a direct thrombin inhibitor
has no effect. Anticoagulation effects appear discordant from
Toll-Like-Receptor 7 mediates inflammatory activity of
Antiphospholipid Antibodies in plasmacytoid dendritic cells
von Landenberg P2, Prinz N1, Lackner LJ1
1
Institute of Clinical Chemistry and Laboratory Medicine, Mainz,
Rheinland Pfalz, Germany; 2Institute of Laboratory Medicine, Olten,
Solothurn, Switzerland
The Antiphospholipid syndrome (APS) is an autoimmune disease char-
acterized by thrombosis, recurrent fetal loss and the presence of anti-
phospholipid antibodies (aPL). Recent data support the idea that the
thrombotic activity in APS patients is attributed to enhanced TNFa
release. To investigate this mechanism more precisely, we employed
two human monoclonal aPL and APS patients’ IgG fractions for sti-
mulation of human plasmacytoid dendritic cells (pDC). All these sti-
muli induced an enhanced expression and secretion of TNFa, caused
by specific upregulation of TLR7-mRNA and protein levels. The direct
dependency between TLR7 upregulation and the increased cytokine
production was confirmed using specific TLR7 agonists and a TLR7
specific inhibitory DNA-oligonucleotide. The TNFa enhancement
could be further increased by TLR7 ligands and nearly completely
neutralized by the TLR7 inhibitor. Furthermore, the addition of
TLR7 ligands to the stimulation assay facilitated maturation process
of pDCs and their ability to activate CD4þ T cells. Since the increased
expression of TLR7 could be confirmed in PBMCs of APS patients we
assume that endogenous stimulation of TLR7 in APS patients and
consecutive elevation of TNFa promote a proinflammatory environ-
ment which can prime T cell activation.
Disclosure: nothing to disclose.
A042
Investigation of role of IL-17 in antiphospholipid syndrom
Popovic-Kuzmanovic D1, Novakovic I2, Trajkovic V3, Stojanovic L1,
Lukovic L2, Aksentijevic I4
1
Medical Center ‘‘Bezanijska kosa’’, Belgrade, Serbia and Montenegro;
2
Institute of Human Genetics, Belgrade, Serbia and Montenegro;
3
Institute of Immunology, Belgrade, Serbia and Montenegro;
4
NIH, Arthritis and Rheumathism Branch, Bethesda, MD
Objectives: Cytokines play an important role in the inflammatory
development and progression of autoimmune diseases. The aim of
present study was to evaluate the role of IL-17 level and IL-17A
gene polymorphism as susceptibility markers for Antiphospholipid
Syndrome (APS). Methods: One hundred Serbian patients with APS
classified according to the Sydney criteria and thirty healthy control
subjects from the same geographic area were recruited. In all subjects
we measured the level of IL-17 in sera by ELISA. Genotyping of
single-nucleotide polymorphism (SNP) in IL-17A gene (rs2275913)
was performed by PCR/SSCP method. Results: The level of IL-17
was significantly higher in bought primary and secondary APS patients
compared with control group (p<0.05). The frequency of rs2275913 G
allele was also higher in APS group than in controls (0.6, 0.69 and 0.45
in primary APS, secondary APS and controls, respectively). The gen-
otype rs2275913 GG was observed in 54%, 52% and 36.6% subjects in

Lupus
 Abstracts

primary APS, secondary APS and control group, respectively.
Conclusion: These results indicate that IL-17 plays an important role
in APS and suggest further studies on IL-17/IL-23 axis involving
immunological and genetic studies.
Disclosure: Nothing to disclose.
A043
Clinical relevance of nitric oxide and nitrative stress in thrombotic
primary antiphospholipid syndrome
Ames PRJ1, Batuca JR2, Ciampa A3, Ferrara F4, Iannaccone L4,
Delgado Alves J2
1
Airedale General Hospital, Steeton, West Yorkshire, United Kingdom;
2
New University of Lisbon, Lisbon, Portugal; 3Moscati Hospital,
Avellino, Italy; 4Cardarelli Hospital, Naples, Italy
Objective: To assess the clinical relevance of nitric oxide (NOx) and
nitrative stress in thrombotic primary antiphospholipid syndrome
(PAPS). Methods: we investigated 46 thrombotic PAPS patients, 21
persistent asymptomatic carriers of antiphospholipid antibodies
(PCaPL), 38 inherited thrombophilia (IT) patients, 33 patients with
systemic lupus erythematosus (SLE) and 29 healthy controls (CTR).
IgG anticardiolipin (aCL), IgG anti-beta-2-glycoprotein I (b2GPI)
crude nitrotyrosine (NT) (indicator of nitrative stress) and high sensi-
tivity C-reactive protein (CRP) were measured by immune-assays.
Plasma nitrite (NO 
2 ), nitrate (NO3 ) and total antioxidant capacity
(TAC) were measured by colorimetric assays. Results: Plasma NO 2
was lowest in PAPS and NO 3 highest in SLE whereas NT was highest
in PAPS and SLE
PAPS PCaPL IT SLE CTR p
509
(VCAM-1). In this study, we measured both surface marker expression
and endothelial microparticle (EMP) release from cultured human
umbilical vein EC (HUVEC) in the presence of IgG from patients
with APS and healthy controls (HC). Methods: 500mg/ml polyclonal
IgG, purified from 28 APS (APS-IgG) and 8 HC (HC-IgG) sera and
confirmed to be endotoxin free, was used to treat fresh HUVEC for
4hours (in triplicate). Cells were stained with fluochrome-conjugated
antibodies to E-selectin, VCAM-1 and tissue factor (TF); cell super-
natant was spun to pellet EMP and stained with AnnexinV (AnV) and
antibodies to a selection of EC surface markers. HUVEC and EMP
were analysed by flow cytometry. EMP were defined as AnV-positive
particles 1.5mm in size, aided by employing 3mm latex beads as a size
cut-off during analysis. Results: 14/28 APS-IgG stimulated expression
of one or more of E-selectin, VCAM-1 or TF, whereas 19/28 stimu-
lated EMP release. Mean expression of cell surface markers did not
significantly distinguish APS-IgG and HC-IgG, while mean values of
EMP generated were significantly increased in the presence of APS-
IgG (2.11.1million EMP/ml) versus HC-IgG (0.70.8 EMP/ml)
(p¼0.0036). EMP also stained for endoglin, VE-cadherin, ICAM-1
and E-selectin. Significantly more E-selectin-positive EMP were gener-
ated in the presence of APS-IgG (mean percentage of total EMP
6.2%4.0) versus HC-IgG (3.4%2.0) p¼0.0172). Conclusions: aPL
can induce EC surface marker expression and EMP generation in vitro.
EMP release, which is increasingly recognised as a biomarker of vas-
cular dysfunction, was a better predictor of aPL-mediated EC activa-
tion, suggesting their potential importance in the APS. The
prothrombotic potential however of these EMP remains to be defined.
Disclosure: Nothing to disclose.
A045

NO2 8.15.6 9.75.1 10.47.3 22.110 30.523.1 <0.0001 b2 Glycoprotein I (b2GPI) Binds Platelet Factor 4 (PF4): Implications
(umol/L) for the Pathogenesis of Antiphospholipid Syndrome
NO3 15.610.3 28.814.5 19.319.6 41.938.5 17.815.1 <0.0001 Sikara MP1, Routsias JG1, Samiotaki M2, Panayotou G2,
(umol/L) Moutsopoulos HM1, Vlachoyiannopoulos PG1
NT (nM) 1.80.6 1.30.4 1.20.7 1.71.0 1.40.7 0.01 1
National University of Athens, Athens, Attiki, Greece;
CRP 0.670.85 0.360.27 0.260.22 1.91.5 0.210.29 <0.0001 2
Al.Fleming BSRC, Vari, Attiki, Greece
(mg/dl)
TAC 10.33 6.92.4 61.5 3.72 7.91.7 0.03 Antiphospholipid syndrome (APS) is an autoimmune thrombophilia
(umol/L)
characterized by arterial/venous thrombosis and/or pregnancy morbid-
ity in the presence of aPL antibodies which mainly recognize b2glyco-
protein I (b2GPI). In order to investigate potential platelet ligands of
In PAPS IgG aCL titre and thrombosis number negatively predicted
b2GPI, platelet membrane proteins from healthy individuals and APS
NO 2 (p¼0.03 and p¼0.001 respectively), arterial thrombosis and patients were passed through a b2GPI-affinity column. Using mass
smoking positively predicted NO 3 (p¼0.05 and p¼0.005 respectively) spectrometry, platelet factor 4 (PF4) appeared as the dominant
and CRP positively predicted NT (p¼0.004). Conclusion: Decreased
b2GPI binding protein. PF4 could bind in vitro, with high affinity,
NO 2 , low grade nitrative stress and low grade inflammation are recombinant b2GPI and the binding was abrogated by soluble
linked phenomena in PAPS and may have implications for thrombosis
b2GPI. Co-precipitation experiments further confirmed this interac-
and atherosclerosis.
tion. In silico molecular docking revealed that PF4 tetramers can
Disclosure: Nothing to disclose. bind two b2GPI molecules simultaneously. Size exclusion chromato-
graphy confirmed that anti-b2GPI antibodies selectively interact with
complexes comprised of (b2GPI)2-(PF4)4. In addition, as demonstrated
A044 by the b2GPI antigenicity evaluation, the reactivity of APS sera was
higher against PF4-b2GPI complex than against b2GPI alone. Upon
Release of endothelial microparticles from human umbilical vein complex formation, anti-b2GPI-b2GPI-PF4 significantly induced pla-
endothelial cells is stimulated by purified IgG from patients with the APS telet p38MAPK phosphorylation and TXB2 production, mainly
Pericleous C1, Clarke LA1, Ioannou Y1, Brogan P1, Latchman D1, through F(ab0 )2 fragments of antibodies. In summary, this study
Isenberg D1, Giles I1, Rahman A1 makes evident that b2GPI forms stable complexes with PF4, leading
1
UCL, London, United Kingdom to the stabilization of b2GPI dimeric structure which facilitates the
antibody recognition. This interaction can probably be involved in
Purpose: Previous studies demonstrated that some aPL activate
the procoagulant tendency of APS.
endothelial cells (EC) in vitro as judged by surface expression of activa-
tion markers such as E-selectin and vascular cell adhesion molecule-1 Disclosure: Nothing to disclose.

Lupus
 Abstracts
510
A046
Binding of phosphatidylserine-targeting antibodies to Junı́n virions
and infected cells
Grant A1, Sbrana E1, Popov V1, Wakabayashi M2, Schlunegger K2,
Freimark B2, Brideau-Andersen A2, Peters C1
1
UTMB, Galveston, TX; 2Peregrine Pharmaceuticals, Tustin, CA
Peregrine Pharmaceuticals, Inc. has developed a broad-spectrum anti-
viral treatment against viral hemorrhagic fevers (VHFs). This therapy
is based upon monoclonal antibodies that target anionic phospholi-
pids, primarily Phosphatidylserine (PS), that are usually present on the
inner leaflet of the plasma membrane in mammalian cells. It has been
hypothesized that most enveloped viruses induce flipping of PS to the
outer leaflet of the plasma membrane during infection. Chimeric and
fully human monoclonal antibodies that target PS, including PGN401
and PGN635, have been shown to bind to a wide variety of virally
infected cells and virions. PGN401 and PGN635 have been shown to
bind to purified Pichinde virions, infected cells, and to protect guinea
pigs upon lethal challenge. Junı́n virus, is serologically distinguishable
from a pathogenic Pichinde, uses a different cellular receptor, and is
the cause of Argentine hemorrhagic fever, which results in 30% mor-
tality in humans. The purpose of this study was to determine whether
PGN401 and PGN635 bind to Junı́n virions and Junı́n infected cells.
Through magnetic bead depletion studies we observed a significant
decrease in number of virions after incubation with the magnetic
beads coated with PGN401 and PGN635, compared to beads coated
with isotype control antibodies. Virions were incubated with PGN401
and PGN635 and labeled with immunogold were observed by TEM.
Purified virus was coated onto plates and PGN401 and PGN635 bind-
ing to the virions was measured compared to control antibodies.
PGN401 and PGN635 binding to virus-infected cells was measured
by confocal immunofluorescent microscopy. In this study, we demon-
strated that antibodies that target PS bind to Junı́n virions by magnetic
bead depletion, immunogold labeling of virions viewed by TEM, and
ELISA. We have also demonstrated binding to infected cells through
IFA. Future studies include pharmacokinetics and efficacy testing in
guinea pigs.
Disclosure: No response indicated.
A047
Anti-serine protease antibodies are distinguishable from antiphospholipid
antibodies in patients with the APS
Turner-Stokes T1, Pericleous C1, Lambrianides A1, Bell K1, Ioannou Y1,
Latchman D1, Isenberg D1, Rahman A1, Giles I1
1
UCL, London, United Kingdom
Purpose: We examined the prevalence of antibodies against procoagu-
lant (thrombin (Thr)) and anticoagulant (activated protein C (APC))
serine proteases (SP) in patients with different clinical manifestations of
the APS, disease and healthy controls (HC) and correlated their pre-
sence with aPL. Methods: Serum was obtained from 61 patients and
22 HC. 32 patients had APS (11 with vascular thromboses (VT) alone,
9 pregnancy morbidity (PM) alone and 12 both VT&PM), whilst
29 patients had SLE but no APS - 16 aPL positive (SLE/aPLþ) and
12 aPL negative (SLE/aPL-). Serum anti-Thr and anti-APC IgG were
measured by ELISA and binding expressed as absorbance units (AU)
by comparison to a human monoclonal anti-Thr/anti-APC IgG, and
positivity was defined as AU greater than the 99th centile of HC.
Serum anti-cardiolipin (aCL) and anti-b2GPI IgG titres were also
Lupus
measured by ELISA. Results: Anti-Thr IgG were positive in
10(31.3%) APS and 5(38.5%) SLE/aPLþ patients, compared to only
1(6%) SLE/aPL- patient and 1(4.3%) HC. Patients with APS had
significantly elevated anti-Thr levels compared to HC (24.1AU vs.
13.6AU, p<0.05). Anti-Thr levels were slightly higher in APS patients
with PM (25.8AU) or VT&PM (26.0AU) compared to patients with
VT alone (20.8AU), and in SLE/aPLþ (31.3AU) compared to SLE/
aPL- (14.6AU) and HC (13.6AU, p<0.05) groups. In contrast, there
was no significant difference in anti-APC IgG levels between any of the
groups studied. There was no significant correlation between anti-Thr
and aCL or anti-b2GP1 titres or between anti-Thr and anti-APC
levels. Conclusions: Our findings show that anti-Thr and anti-APC
do not clearly distinguish patients with the APS from SLE/aPLþ
patients and that these anti-SP are distinct from other aPL. Despite
the homologous catalytic sites shared by these two SP, our results
suggest that anti-Thr do not necessarily cross-react with APC.
Disclosure: Nothing to disclose.
A048
In Vivo Involvement Of Apolipoprotein E Receptor2 (apoER20 ) On
Thrombogenic Effects Of A Dimer Of b2glycoprotein I (b2GPI)
Aguilar-Valenzuela R1, Romay-Penabad Z1, Urbanus R2, Shilagard T1,
Papalardo E1, Vargas G1, Derksen R2
1
University of Texas Medical Branch, Galveston, TX; 2University
Hospital Utrecht, Utrecht, The Netherlands
Background: In vitro, dimeric b2GPI mimics the effects of b2GPI/anti-
b2GPI antibodies complexes on target cells, binds to apoER2’ in plate-
lets, increases their adhesion to collagen and induces the activation of
endothelial cells. Those effects can be diminished by soluble binding
domain I of apoER2’ (sBD1). However, the role of dimeric b2GPI and
apoER20 in vivo remains unknown. Methods: apoER2/-or apoER2þ/þ
mice were injected with dimeric b2GPI or with control. Then, several
procedures were done in each mice: a) thrombus size, b) tissue factor
(TF) function in homogenates of carotid arteries and in peritoneal
macrophages. Results: Dimeric b2GPI induced significantly larger
thrombi and increased TF activity in apoER2þ/þ mice when compared
to apoER2/-. Those effects were significantly diminished in apoER2þ/þ
mice treated with dimeric b2GPI þ sBD1
Thrombus TF in carotid TF in peritoneal
size artery macrophages
Mice/treatment (m2) (pM/ mg protein) (pM/ mg protein)
1. apoER2þ/þ/ Dimer *37571624 *34624 *18736
2. apoER2/-/ Dimer 1365502 17.59.1 13.93.0
3. apoER2þ/þ/ **533148 **12315 **2.11.7
Dimer þ sBD1
*statistically significant different from 2. ** statistically significant different
from 1.
Conclusions: Altogether these data show that prothrombotic properties
of dimeric b2GPI are mediated through apoER20 in vivo contributing
to our understanding of the pathogenesis of anti-b2GPI antibodies
in APS.
Disclosure: Nothing to disclose.
 Abstracts

511
A049 More specifically, we found that interneurons (Basket cells) in the
hippocampus were intensely stained for IgG in APS mice. This staining
Apolipoprotein E Receptor (apoER2) Mediates Thrombogenic Effects was highly specific for the 5 ePAS brains stained compared to 5 adju-
Of Antiphospholipid (aPL) Antibodies vant immunized control mice. Both in vitro and in vivo binding patterns
Romay-Penabad Z1, Shilagard T1, Vargas G1, Aguilar-Valenzuela R1, are compatible with the neurobehavioral effects: The Basket cell inter-
deGroot P2, Pierangeli S1 neurons are inhibitory and secrete GABA. We suggest that a loss of
1
University of Texas Medical Branch, Galveston, TX; 2University such neurons can account for the hyperactive behavior in eAPS mice
Hospital Utrecht, Utrecht, The Netherlands and possibly also account for the learning and memory deficit.
Furthermore, similar selective loss of inhibitory neurons in the brain
Background: It has been suggested that b2GPI-APL antibody complex may account for the increased incidence of epilepsy associated with
binds to apoER2 in cell surface, inducing activation of target cells. APS in humans. Thus, the eAPS model provides a unique tool to
Moreover, a soluble binding domain of apoER2 (sBD1) inhibits the define the anatomical, physiological, and signaling effects of the aPL
binding of b2GPI to apoER2 in vitro. Here, we examined the involve- in vitro and in vivo.
ment of apoER2 and the effects of sBD1 on aPL-mediated pathogenic
effects in vivo in mice. Methods: Several procedures were done in Disclosure: Nothing to disclose.
apoER2/-or apoER2þ/þ mice injected with aPL (IgG-APS) or with
control IgG (IgG-NHS) or with IgG-APS þ sBD1 or IgG-NHS þ A051
sBD1: a) thrombus size, b) TF function in homogenates of carotid
arteries and in peritoneal macrophages, and c) TF (macrophages) Protein Z Gene Polymorphisms in Systemic Lupus Erythematosus
expression using quantum dot nano crystals and two-photon excitation Taib S1, Mant C2, Cason J2, Bertolaccini ML1, Hughes GRV1,
laser scanning microscopy. Results: Significantly larger thrombi and Khamashta MA1
increased TF activity in carotid artery homogenates and in peritoneal 1
The Rayne Institute, King’s College London School of Medicine,
macrophages in apoER2þ/þ mice when compared to apoER2/-mice London, United Kingdom; 2Department of Infectious Diseases, King’s
treated with aPL. Importantly, thrombogenic effects of IgG-APS were College London School of Medicine, London, United Kingdom
significantly abrogated in apoER2þ/þ mice treated with aPL þ sBD1.
Background: Low ProtZ levels have been reported in patients with
TF in TF in TF
thromboembolic diseases, adverse obstetric history (AOH) and in
carotid peritoneal expression patients with aPL and antibodies to ProtZ. Polymorphisms in the
Thrombus artery macrophages (fluorescence ProtZ gene may affect ProtZ levels and thus play a role in thrombosis.
size pM/ mg pM/ mg intensity, We investigated the prevalence and clinical significance of ProtZ gene
Mice/treatment (m2) protein protein AU) a-13g and g79a polymorphisms in patients with SLE. Patients and
Methods: 90 SLE patients (mean age 44  12.3 years; disease duration
apoER2þ/þ/aPL *40941037 *21.48.4 *10.11.2 *30.14.2 11.7  8.5 years; 87 female), and 49 age and sex matched healthy
apoER2/-/aPL 1675781 14.54.0 5.91.5 15.08.9 individuals were genotyped for a-13g and g79a polymorphisms by
apoER2þ/þ/ **665173 **14.81.0 **1.51.2 **18.74.8 PCR-RFLP analysis and DNA sequencing. IgG and IgM antibodies
aPL þ sBD1
to ProtZ were tested by an in-house ELISA. Plasma ProtZ was mea-
sured by a sandwich ELISA. Results: The prevalence of a-13g and g79a
*statistically significant different from 2. ** statistically significant different
genotypes was not significantly different between SLE patients and
from 1.
healthy controls (18.9% vs. 8.3%, p ¼ 0.1 for a-13g AG/GG; 40%
vs. 32.7%, p ¼ 0.39 for g79a GA/AA). These polymorphisms did not
Conclusions: Altogether these data show that apoER2’ is a mediator of differ between SLE patients with aPL, thrombosis, AOH or antibodies
aPL thrombogenic effects in vivo.
to ProtZ when compared to those without. Plasma concentration of
Disclosure: Nothing to disclose. ProtZ was higher in patients with rare type a-13g (GG) variant com-
pared to those with wild type variants (a-13g [GG]: 2.11  0.01 mg/ml
[95% CI 1.98-2.24] vs. a-13g [AA]: 1.43  0.74 mg/ml [95% CI 1.08-
A050 1.77], p ¼ 0.0006). The prevalence of a-13g (AA) variant was asso-
ciated with thrombosis in 10/28 patients in whom ProtZ levels were
Selective involvement of inhibitory neurons in the pathogenesis of neural measured (p ¼ 0.04). Conclusion: ProtZ gene polymorphisms are not
injury in experimental Antiphospholipid Syndrome associated with SLE, APS, or ProtZ antibodies. The a-13g AG/GG
Katzav A1,2, Menachem A1,2, Chapman J1,2 variant may be protective against thrombosis by predisposing to
1
Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel; 2Tel Aviv elevated ProtZ levels.
University, Ramat Aviv, Tel Aviv, Israel
Disclosure: Maria Laura Bertolaccini is funded by the Louise Gergel
The antiphospholipid syndrome (APS) is characterized by central Fellowship.
nervous system (CNS) dysfunction associated with antibodies to a
complex of phospholipids and b2-glycoprotein I (b2-GPI) causing A052
hyper-coagulation and immune mediated neurodegenerative mechan-
isms. Mice immunized with b2-GPI develop systemic and CNS mani- Blood Brain Barrier Dysfunction and Central Nervous System Pathology
festations of experimental APS (eAPS). The aim of the present study in Experimental Antiphospholipid Syndrome
was to evaluate the pathogenic mechanisms involved, in particular Katzav A1,2, Menachem A1,2, Chapman J1,2
1
direct effects of antiphospholipid antibodies (aPL) on the brain. Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel; 2Tel Aviv
Histological stainings for IgG binding were performed in vitro on University, Ramat Aviv, Tel Aviv, Israel
normal brain slices and in vivo on APS and control brain slices. aPL
from APS mice bind in vitro to specific brain structures, most of them Circulating antiphospholipid antibodies (aPL) are associated with cen-
white matter tracts in the limbic system. Specific accumulation of aPL- tral nervous system dysfunction in antiphospholipid syndrome (APS)
IgG in brain neuronal cells was demonstrated in vivo in APS mice. patients and in a mouse model of APS. The well-defined behavioral

Lupus
 Abstracts
512
and cognitive effects of experimental APS (eAPS) in mice offer a
unique model in which to study the pathogenic mechanisms involved,
in particular direct effects of aPL on the brain. An important step in
the pathway leading to direct antibody effects on the CNS is their
passage through the blood–brain barrier (BBB) of which brain vascu-
lar endothelial cells are the principal component. In the present study
we assessed the BBB function in eAPS. Integrity of BBB was quantita-
tively evaluated by measurement of extravasated Evans blue dye in
brain parenchyma by both fluorescence microscopy and by spectro-
scopy of dye extracted from brain homogenates. All mice underwent
perfusion through the heart to minimize intravascular dye in the brain.
We found that BBB disruption was enhanced in eAPS (n¼5) compared
to naı̈ve mice (n¼5). Significant disruption of the BBB was also found
in adjuvant immunized mice (n¼5) compared to naı̈ve mice. IgG con-
tent in the brain homogenates was however higher in eAPS compared
to both control groups and histological studies confirmed significantly
higher levels of IgG accumulated in cells compatible with neurons
in vivo in eAPS. These results indicate that although the BBB is dis-
rupted in both eAPS and adjuvant controls, only in eAPS this asso-
ciated with IgG accumulation in the brain, possibly due to the presence
of high levels of brain binding IgG in the eAPS serum. These results
suggest that lowering the levels of aPL is an important therapeutic goal
in APS.
Disclosure: Nothing to disclose.
A053
Further analyses of antiphospholipid antibodies that impair antithrombin
inactivation of procoagulant factors
Meifang Wu M1, FitzGerald J1, Grossman J1, Hahn B1, Chen P1
1
UCLA, Los Angeles, CA
This study is to further characterize the antiphospholipid antibodies
(aPL) that react with procoagulant factors (thrombin, FXa and FIXa)
and hinder inactivation of the target factors by antithrombin (AT).
First, we used fibrinogen to analyze effects of the thrombin-reactive
aPL on hindering AT inactivation of thrombin. Second, we developed
an activated-partial-thromboplastin-time (APTT)-based assay to study
the overall effects of the reactive aPL on AT inactivation of procoa-
gulant factors in plasma. In addition, to avoid the problem that some
reactive aPL possess lupus anticoagulant activity (that prolong clotting
time) and thus could obscure aPL-mediated impaired AT function (in
shortening coagulation time), we assessed the effect of each test aPL on
AT function by comparing the clotting time of the aPL in plasma
containing heparin (with AT function) with that in plasma containing
no heparin (without AT function). Then we divided the clotting time
with heparin by the clotting time without heparin, and expressed the
results as the clotting ratio. The results showed that three of the eight
patient-derived IgG monoclonal aPL (i.e., B2, CL15 and CL24) at the
physiological concentration hindered AT inactivation of thrombin.
Moreover, IgG was purified from blood samples of three patients
with APS, and two of these IgG preparations similarly hindered AT
inactivation of thrombin, confirming the pathogenic significance of
Lupus
AT-interfering anti-thrombin antibodies in the antiphospholipid syn-
drome (APS). At the global level in plasma, two of the eight mono-
clonal aPL (i.e., B2 and CL24) at the physiological concentration
reduced clotting ratios from 250% for normal human IgG to 144 %
(p < 0.01) and 171% (p < 0.05). Moreover, two of the above three
polyclonal IgG also reduced clotting ratio to 153% and 145%, as
compared with a clotting ratio of 175% for normal human IgG.
These reduced clotting ratios reflected that these monoclonal and poly-
clonal aPL accelerated the clotting time by impairing AT inactivation
of procoagulant factors in plasma. Taken together, these results show
the AT-interfering IgG aPL are present in some APS patients and
could contribute to thrombosis in such patients.
Disclosure: Nothing to disclose.
A054
Prolactin: does it have a role in the pathogenesis of antiphospholipid
syndrome?
Praprotnik S1, Agmon-Levin N2, Porat-Katz B3, Blank M2, Rozman B1,
Meroni P4, Cervera R5, Miesbach W6, Scyper-Kravitz M2,
Stojanovich L7, Shoenfeld Y2
1
University Clinical Center, Ljublijana, Slovenia; 2Sheba Medical
Center, Tel-Aviv, Tel-Aviv, Israel; 3University of Jerusalem, Jerusalem,
Israel; 4University of Milan, Milan, Italy; 5Hospital Clinic, Barcelona,
Catalonia, Spain; 6University of Frankfurt, Frankfurt, Germany;
7
University of Belgrade, Serbia, Serbia and Montenegro
Background and Aim: Increased levels of serum prolactin were reported
in patients with various autoimmune diseases and have been associated
with lupus disease activity. Currently, there is lack of data regarding
hyperprolactinemia (HPRL) in patients with the antiphospholipid syn-
drome (APS). The aim of the present study was thus to evaluate the
prevalence and clinical significance of HPRL in APS. Methods: 172
European patients with APS and 100 geographically and sex-matched
healthy controls were included, none had obvious causes of HPRL. All
patients underwent clinical assessment for disease manifestations in
addition to laboratory assessment for serum prolactin, other biomar-
kers of autoimmune diseases and for common infectious agents. The
tests were performed on the LIAISONÕ Analyser (DiaSorin, Sallugia
Italy). Results: Hyperprolactinemia (HPRL) was detected in 21/172
APS patients and 0/100 controls. This significant difference was present
in both genders and was obvious even after sub-grouping the patients
in primary and secondary APS. When clinical features were compared,
HPRL was associated to reproductive failure, including early and late
pregnancy loss, as well as intrauterine growth retardation. HPRL was
negatively related to arthralgias, venous thrombosis, pulmonary
thrombosis, pulmonar hypertension and neurological manifestations.
HPRL was not related to infections or other biomarkers for autoim-
mune diseases. Conclusion: These data indirectly imply that prolactin
may play a role in the pathogenesis of APS, especially APS related
reproductive failure.
Disclosure: Nothing to disclosure.
 Abstracts

513

Abstracts presentations Thursday April 15th, 2010

B055 unexplained pregnacy loss (UPL) both, recurrent miscarriage (RM)

Maternal and fetal outcome in antiphospholipid syndrome after stroke
Alba P1, Alvarez A1, Palena C1, Ropato L1, Guarneri F1, Buteler MM1,
Crespo Roca F1
1
Materno Neonatal hospital, Córdoba, Argentina
Introduction: Women with antiphospholipid syndrome (APS) can
experience maternal and fetal complications. Although it is widely
accepted that antithrombotic treatment and a close follow up improve
outcome of pregnancies of APS, a controversy exists regarding the
timing of stroke in pregnancy. Objective: We prospectively evaluated
12 pregnancies in 9 women with diagnosis of APS with previous stroke
according to Sydney criteria who attended Materno Neonatal Hospital
since to January 2007 to December 2009. All the patients received
treatment with aspirin plus anticoagulation dose of low molecular
weight heparin (LMWH) delivery. The timing and mode of delivery
was assessed individually. Maternal complications evaluated were:
death, arterial thrombosis (AT), venous thrombosis (VT),
Preeclampsia (PE) and others. Fetal outcome was considered as: live
birth, fetal weight at birth and neonatal death. Results: Mean age of
patients was 29 years old. Mean previous pregnancies were 2 and a
mean live birth 1. Mean time from previous stroke to the pregnancy
was 44 months (6-78). Only 1 patient had AT, and 1patient PE and
there were not VT or maternal death. The caesarean section rate was
81%. Main indications were: previous cesarean (5), Failure induction
(1), and fetal distress (4). Live birth rate was 90 % and 31% had low
birth weight without neonatal death. Conclusion: Successful maternal
and fetal outcome can be achieved in APS pregnancies with previous
stroke after 6 months of the stroke event.
Disclosure: Nothing to disclose.
B056
and fetal loss (FL). Patients and Methods: 89 pregnant women divided
in two groups. Study group: 51 women (53 cases) with UPL and 38
healthy pregnant control group. 20 healthy non-pregnant women acts
as a control. cMP were analyzed by means of flow cytometry methods.
aPL detection were analysed with standard ELISA methods and coa-
gulometric assays (LA). Results: Platelet (p¼0.05) and endothelial
(p¼0.002) cMP increased in healthy pregnant women. PE/FGR
whole group showed an increase of endothelial (p¼0.032) and CD41-
negative (p¼0.001) but not CD41þ cMP. Comparing PEFGR vs.
controls, increasing levels of endothelial (p¼0.024) and CD41-
(p¼0.001) cMP were found. Comparing FGR to controls, slight differ-
ences in leukocyte cMP were obtained. Comparing PE group to
isolated FGR, significant CD41- (p¼0.002) and endothelial
(p¼0.080) cMP decrease were obtained. Analyzing differences between
cMP in PEFGR according its aPL positivity, a decrease in leukocyte
and endothelial cMP levels were found. Conclusions: Normal preg-
nancy increases levels of platelet and endothelial cMP. When compar-
ing UPL, RM or FL groups with controls, increased number of
cMP were not observed. On the contrary, significant endothelial cMP
decrease in UPL and RM were obtained. No differences in cMP number
were seen when comparing aPLþ vs. aPL - populations. Complement
levels were decreased in aPLþ subgroup.
Disclosure: No response indicated.
B057
Circulating cell-derived microparticles in severe Preeclampsia, HELLP
syndrome and in Fetal Growth Restriction
Zarzoso C1, Garcia-Ruiz I1, Lopera A1, Farran-Codina I1,
Alijotas-Reig J1
1
Vall d’Hebron University Hospital. UAB, Barcelona, Spain

Circulating cell-derived microparticles and antiphospholipid antibodies in Aim: To analyze type and number of different circulating, cell-derived
unexplained pregnancy loss microparticles (cMP) in severe PE and FGR. Patients and Methods:
Zarzoso C1, Garcı´a-Ruiz I1, Farran-Codina I1, Alijotas-Reig J1 cMP of 70 cases of severe PE/HELLP syndrome or FGR were
1 compared to cMP of 38 healthy pregnant women.20 healthy non-preg-
Vall d’Hebron University Hospital. UAB, Barcelona, Spain
nant women were used as controls. cMP were analyzed by means
Aim: To analyze type and number of different circulating, cell-derived of flow cytometry methods. Results are given as total (ANXA5þ),
microparticles (cMP) and antiphospholipid antibodies (aPL) in platelet (CD41þ), leukocyte (CD45þ), endothelial (CD144þ;

Table 1

aPL UPL Control p FL Control p RM Control p

LA 5/48 10.4% 0/38 0 0.040 3/20 15% 0/38 0% 0.014 4/35 11,4% 0/38 0% 0.032
aCL IgG 8/48 16,7% 1/38 2,6% 0.034 5/21 23,8% 1/38 2,6% 0.010 5/34 14,7% 1/38 2,6% 0.064
IgM 7/48 14.5% 1/38 2,6% 0.054 4/21 19.04% 1/38 2,6% 0.025 5/34 14,7% 1/38 2,6% 0.064
IgG/IgM 9/48 18.7% 1/38 2,6% 0.019 5/21 23.8% 1/38 2,6% 0.007 6/34 17,6% 1/38 2,6% 0.031
ab2GPI-ab IgG 6/43 14% 1/38 2,6% 0.070 2/19 10.5% 1/38 2,6% 0.188 4/29 13,8% 1/38 2,6% 0.085
IgM 4/43 9,3% 1/38 2,6% 0.213 3/19 15.7% 1/38 2,6% 0.056 1/29 3,44% 1/38 2,6% 0.844
IgG/IgM 7/43 16,2% 1/38 2,6% 0.039 3/19 15.7% 1/38 2,6% 0.066 4/29 13,8% 1/38 2,6% 0.085

aCL IgG/IgM: anticardiolipin antibodies IgG/IgM isotypes; anti-b2GP-I-ab IgG/IgM isotypes: anti b2glycoproteinI antibodies; aPL: antiphospholipid
antibodies; LA: lupus anticoagulant.

Lupus
 Abstracts
514
CD31þ/CD41-), and CD41-negative c-MP per mL of plasma. Absolute
number of microparticles was calculated by means of the specific for-
mula. Results: Platelet (p¼0.05) and endothelial (p¼0.002) cMP were
increased in healthy pregnant vs. non-pregnant healthy women. PE/
FGR whole group showed an increase of endothelial (p¼0.032) and
CD41-negative (p¼0.001) but not CD41þ cMP. When comparing
PEFGR vs. controls increasing levels of endothelial (p¼0.024) and
CD41- (p¼0.001) cMP were also seen. Comparing FGR with controls,
slight differences in leukocyte cMP were obtained (p¼0.006).
Comparing PE group with isolated FGR, significant decrease of
both CD41-negative (p¼0.002) and endothelial (p¼0.080) cMP levels
were obtained. When we analyzed the differences between cMP in the
study group according its positivity for aPL, a decrease in leukocyte
(p¼0.07) and endothelial (p¼0.06) cMP levels were observed, specially
in LA þ women. Conclusions: Normal pregnancy increases levels of
platelet and endothelial cMP. Neither total nor platelet cMP increase
were seen in any study groups. Increasing CD41 negative and endothe-
lial cMP levels were seen in PE groups but isolated FGR group. PE or
FGR women positive for aPL showed lower leukocyte and endothelial
cMP levels than aPL negative.
Disclosure: No response indicated.
B058
Antiphospholipid Syndrome (APS) in Pregnancy: Results from the
Serbian National APS Registry
Stojanovich L1, Djokovic A1, Damnjanovic´-Pazin B2, Mikovic Z2,
Marisavljevic D1, Elezovic I3
1
Clinical Center Bezanijska kosa, Belgrade, Serbia and Montenegro;
2
Clinic of Ginecology and obstetrics ‘‘Narodni Front’’, Belgrade, Serbia
and Montenegro; 3Clinical Center of Serbia, Belgrade, Serbia and
Montenegro
Introduction: Antiphospholipid Syndrome (APS) patients suffer from
venous or/and arterial thrombosis and repeated fetal losses. APS may
manifest itself as a primary disease (PAPS), or as a secondary disease,
coexisting most commonly with the Systemic Lupus Erythematosus
(SLE). Patients and Methods: The project started in January 2006.
Data currently comprises total of 256 patients: 162 PAPS patients
(94 female and 68 male; mean age 45.2 þ 12.9 years) and 94 SLE
patients with secondary APS (83 female and 11 male; mean age 43.1
þ 15.4 years); aPL analysis includes analysis of aCL (IgG/IgM),
b2GPI, and LA. Total of 156 pregnant women: 95 PAPS patients
and 61 SLE patients were analyzed. In all patients echocardiography
study was performed and data considering acute myocardial infarc-
tion, unstable angina, coronary artery bypass grafting (CABG) or
percutaneus coronary artery angioplasty (PTCA) as well as manifesta-
tions of chronic or acute heart failure were also collected. All patients
were treated with anticoagulant and antithrombocyte therapy (aspirine
with low molecular fractioned heparin). Results: Echocardiography
changes were not revealed in patients with PAPS, only one patient in
SLE group had valvular changes and the same patient had sympthoms
of unstable angina (p¼0.391). The results of serology testing for aPL
between PAPS and SLE, showed no statistically significant differences.
Conclusions: According to data of the Serbian National APS Registry
women with antiphospholipid syndrome and adequate treatment do
not experience significant cardiac manifestations as well as echocardio-
graphy changes during pregnancy.
Disclosure: Nothing to disclose.
Lupus
B059
Pregnancy outcome in APS patients
Ljudmila Stojanovich L1, Damnjanovic Pazin B1, Mikovic Z1,
Marisavljevic I1
1
University Medical Center ‘‘Bezanijska Kosa’’, Belgrade, Serbia and
Montenegro
Introduction: APS patients suffer from venous and/or arterial throm-
bosis and repeated fetal losses in the presence of antiphospholipid
antibodies (aPL). APS may manifest it self as a primary disease
(PAPS), or as a secondary disease, coexisting most commonly with
the Systemic Lupus Erythematosus (SLE). Goals: Demonstration of
the initial results from the Serbian National APS Registry, related to
implications of APS to the pregnancy outcome. Patients and Method:
The project started in January 2006. Data currently comprises total of
256 patients: 162 PAPS (94 female; 68 male; mean age 45.2þ12.9 y.); 94
SLE patients with secondary APS (83 female;11 male; mean age
43.1þ15.4 y.). Currently data include total of 156 women who had
positive pregnancy anamnesis (95 PAPS; 61 SLE). APS analysis
include detection of aCL (IgM/IgG); ab2GPI (IgM/IgG) and LA.
Results: Among 156 women with positive pregnancy anamnesis, 88
of them (56,4%) had fetal loss(p¼0.034). The statistically significant
difference (p¼0.002) was observed comparing 60 (63.2%) PAPS
patients with 28 (45.9%) SLE patients, respectively. The results of
serology testing for aPL between PAPS and SLE showed no statisti-
cally significant difference:58% PAPS with positive aCL-IgG had fetal
loss vs. 53,6% SLE aCL-IgG positives with fetal loss (p¼0,593).
Results of aCL-IgM in both groups of women were similar (PAPS
vs. SLE:50.8% and 60.0% of fetal losses(p¼0.259)), so as ab2GPI-
IgG and IgM (52% PAPS vs. 64,3% SLE:p¼0,138))and LA (54,3%
PAPS vs. 58.7% SLE:p¼0.584). Conclusion: The Serbian National
APS Registry allowed us to ascertain a significantly increased incidence
of fetal losses in PAPS patients, compared to patients with secondary
APS, as far as statistically increased risk for fetal losses, comparing to
good fetal outcome among all APS women. Comparative analysis of
aPL did not show a statistically significant difference between fetal
losses and the type of antiphospholipid antibodies.
Disclosure: Nothing to disclose.
B060
Obstetric outcome in patients with antiphospholipid syndrome (aps) and
in patients with low titer antiphospholipid antibodies
Barros V1, Bortolotto MR1, Oliveira A1, Baptista F1, Zugaib M1
1
Clinics Hospital, University Of Sao Paulo, Sao Paulo, Brazil
Background: The new criteria to define patients with APS exclude
many patients with clinical criteria and persistent low titer antipho-
spholipid antibodies. Objectives: to compare the outcome of the preg-
nancies of patients with new criteria and those with low titer
anticardiolipin antibodies. Methods: APS group comprised 43 patients
and the APS low titer (LT) group had 73 patients. Anticoagulation:
enoxaparin in 98% of cases. Results: APS group had 47 pregnancies/
47 fetuses. APS-LT group had 85 pregnancies/ 86 fetuses. Patients
older than 35 year were more frequent in group APS-LT (22% vs.
10% APS group). Both groups had a similar poor obstetric past his-
tory (32% vs. 40% APS-LT). Recurrent abortions were more frequent
in APS-LT(48% vs. 18% APS). APS-LT group had a higher incidence
of another thrombophilia (27% vs. 9%) but the finding of abnormal
levels of lipoprotein-A were more frequent in the SAF group(23% vs.
12%). The reporting of a previous DVT was similar in both groups
(32%), although the incidence of DVT during pregnancy was higher in
the SAF group(18% vs. 9%). Another risk factors as chronic

hypertension, diabetes, smoking, obesity (BMI > 30) were similar in
both groups. Higher doses of LMWH in the third trimester were neces-
sary in the APS group (44% vs. 20%). The pregnancies outcomes were
worse in the APS-LT group: restricted fetal growth(APS 4% vs. APS
LT 21%), fetal death(4% APS-LT vs. 0% APS), premature birth
< 32nd week(APS-LT 7% vs. 0% APS). The incidence of placental
histologic abnormalities was similar in both groups (16%).
Conclusions: low titer anticardiolipin pregnant patients had a worse
obstetric outcome, probably associated with low doses of heparin pre-
scribed. New protocols of anticoagulation need to be established.
Disclosure: Nothing to disclose.
B061
Antiphospholipid syndrome and detachment of chorion
Makatsaria A1, Kuneshko N1, Samburova N1
1
The I.M.Sechenov Moscow Medical Academy, Moscow, Russian
Federation
As is known, the antiphospholipid syndrome causes obstetric compli-
cations: pregnancy loss, premature detachment of placenta, gestosis.
We studied a role of antiphospholipid antibodies (APA) at patients
with detachment of chorion. It has been examined 157 pregnant
women with detachment of chorion on term from 4 till 12 weeks of
pregnancy. 126 (80.3%) patients had a syndrome of fetus loss, 40
(25.5%) - premature detachment of placenta in the anamnesis. To all
patients it was performed hemostasis research, detection of antipho-
spholipid antibodies and genetic thrombophilia. Results: Circulation
of APA has been detected at 102 (64.9%) patients. APA included lupus
antibodies (LA) (37.6%), anticardiolipin antibodies (39.5%), antibo-
dies to cofactors (26.8%). 86 (54.8%) women had a combination of an
APA circulation and genetic thrombophilia. All patients had high
levels of markers of thrombophilia (RAR, D-dimer). Treatment
included antihaemorrhagic (Rraneksam) and antithrombotic medicine
(LMWH), vitamins, antioxidants. Pregnancy has been kept at 94.3%
of women. Conclusion: Antiphospholipid antibodies can cause a
detachment of chorion. We recommend to determine the circulation
of APA at all patients with thrombotic and obstetric complications in
the anamnesis. Detection of APA permits to use antithrombotic drug
in treatment and preventive detachment of chorion that improve effi-
ciency of therapy.
Disclosure: No response indicated.
B062
The prevalence of antiphospholipid antibodies 3-16 years after a
pregnancy-related venous thrombosis
Bergrem A1, Jacobsen EM1, Skjeldestad FE1, Flem Jacobsen A1,
Morten Sandset P1
1
Oslo University Hospital, Ullevaal, Oslo, Norway
Background/Methods: In this population-based case-control study we
explored the association of antiphospholipid antibodies (APAs) with
pregnancy-related venous thrombosis (VT). From 1990 through 2003,
615 pregnant women were identified at 18 hospitals in Norway with a
diagnosis of first time VT. In 2006, 531 of 615 eligible cases and 1092 of
1230 eligible controls were invited to further investigations. The final
study population completed blood sampling and returned a compre-
hensive questionnaire during 2006, 3-16 years after index pregnancy.
Lupus anticoagulant (LA) was identified by using two validated in-
house Lupus Ratio tests, APTT-based and RVVT-based, respectively.
Anticardiolipin (aCL) IgG and IgM were analyzed by validated in-
house standards. Anti-b2 glycoprotein 1 (anti-b2GP1) IgG and IgM
Abstracts
515
were determined with commercial ELISA kits. The 99 percentiles of
the control group were used as upper reference limits. Results: The
final study-population comprised 313 cases and 353 controls. 29
(9.3%) of the cases and 24 (6.8%) of the controls had at least one
positive test for APAs (OR 1.4, 95% CI 0.8-2.5). No differences were
found in prevalence of LA, anti-b2GP1, and aCL antibodies among
cases and controls. Nine cases (2.8%) and no controls had more than
one positive test for APAs. There were too few APA positive women in
order to do meaningful interaction analysis of clinical risk factors and
APAs in association to VT. Conclusions: The prevalence of positive
tests for APAs, 3-16 years after index pregnancy, were not significantly
different between women with a history of pregnancy-related first time
VT and controls. The presence of more than one positive test for
APAs was weakly associated with a history of pregnancy-related first
time VT.
Disclosure: Nothing to disclose.
B063
Obstetrics Outcomes Associated with an Increased Level of
Lipoprotein-A (Lp-A) in Thrombophilic Patients
Barros V1, Baptista F1, Bortolotto MR1, Igai A1, Oliveira A1,
Zugaib M1
1
Clinics Hospital University of Sao Paulo, Sao Paulo, Brazil
An increased level of Lp(a) can be a new marker for adverse pregnancy
outcome and/or thromboembolism risk during pregnancy. We studied
69 patients diagnosed with deep vein thrombosis (current or past) and /
or thrombophilia. They were divided into 2 groups: patients with
increased Lp(a)(Group A), and patients with negative values (Group B).
Group A(GA) had 47 patients and Group B(GB) had 22 patients.
Patients black or brown equaled 42% of the GA, compared with
22% of GB population. A past history of adverse pregnancy outcome
was present in 36.2% of GA and recurrent abortions in 12%, with
incidences respectively 18% and 0% in GB. The prevalence of deep
venous thrombosis (DVT) was similar in both groups (42.5% and
54.5%), but 25.5% of cases of GA occurred in previous pregnancies
compared to 9% in GB. Another significant finding was the presence
of recurrent DVT in 10.6% of GA patients compared with 4.5% of the
GB. Smoking, obesity (BMI>30) and patients older than 35 years had
similar results in both groups. The antiphospholipid antibodies syn-
drome (APS) was more frequent in GA(21.3%) than in GB(13.6%),
but the prevalence of genetic thrombophilia was similar in both
groups. Abnormal placental findings occurred in 27.6% in GA, against
13.6% in GB.Conclusions: these findings suggest that elevated Lp(a) is
a new thrombophilic risk factor in pregnancy.
Disclosure: Nothing do disclose.
B064
Outcome of the pregnancies of patients with Antiphospholipid Syndrome
(APS) and combined thrombophilia (ct)
Barros V1, Baptista F1, Bortolotto MR1, Oliveira A1, Zugaib M1
1
Clinics Hospital University of Sao Paulo, Sao Paulo, Brazil
Background: There are no treatments protocols of anticoagulation for
APS associated with other thrombophilias. Objectives: To compare the
outcome of the pregnancies of patients with APS and CT. Methods:
APS group comprised 116 patients and the CT group had 22 patients.
Anticoagulation: enoxaparin in 90, 15% and 96% of cases respectively.
The thrombophilia investigated were: Protein S deficiency (52%),
Prothrombin Mutant (24%), Leiden Factor (8%), Protein C and
Antithrombin deficiency (4% each one). Results: APS group had 132
Lupus
 Abstracts
516
pregnancies/ 134 fetuses. CT group had 25 pregnancies/26 fetuses.
Both groups had a similar poor obstetric past history (41.6% vs.
36% CT). Recurrent abortions were more frequent in APS group
(28.8% vs. 16% CT). The reporting of a previous DVT was similar
in both groups (25%), although the incidence of DVT repetition was
higher in the APS group (12% vs. 4% CT), as well as stroke (9% vs.
0% CT).Diabetes were more frequent in CT group (20% vs. 10%
APS). Other risk factors as chronic hypertension, smoking, obesity
(BMI > 30), patients older than 35 year, were similar in both
groups. History of hypertension and hypertensive disorders of preg-
nancy were more frequent in APS group(20,5% and 6,8%), compared
to the CT group(4% and 0%), although the incidence of family history
of thrombosis was higher in CT group (28% vs. 13.6% APS) The
pregnancies outcomes were worse in the APS group: fetal death(3%
APS vs. 0% CT), premature birth < 37nd week(APS 35.6% vs.
27% CT), oligohydramnios(APS2,2% vs. 0% CT), fetal distress
(APS12,1% vs. 4% CT), eclampsia (1,5% vs. 0% CT). The incidence
of placental histological abnormalities was higher in the APS group,
especially infarction (6% vs. 0%CT) and massive intervilous fibrin
deposition (4.5% vs. 0% CT). Conclusions: The obstetrics worst out-
comes and placental histological abnormalities were more frequent in
the APS group, probably due to more aggressive anticoagulation in the
CT group.
Disclosure: Nothing to disclose.
B065
Hypocomplementemia and obstetric complications in women with
Primary APS: An italian multicentric study
Ziglioli T1, Mosca M2, Ramoni V3,4, Iuliano A5, Gerosa M6, Andreoli L1
1
Brescia University, Brescia, Italy; 2Pisa University, Pisa, Italy;
3
IRCCS Policlinico San Matteo, Pavia, Italy; 4Ospedali Riuniti di
Bergamo, Bergamo, Italy; 5Siena University, Siena, Italy; 6Ist.
Auxologico Italiano, Milano, Italy
Purpose: To evaluate the association between complement (C) serum
levels and pregnancy complications in women with PAPS. Methods: 41
pregnancies in 29 women with PAPS (cases) and 43 pregnancies in 35
women with Sjogren S.or UCTD (controls) were studied. Levels of C3
and C4 were tested by nephelometry before and during pregnancy.
Cases and controls were subdivided in subgroups according to C
level. Subgroups were tested by 2 test for differences in obstetric
complications prevalence. Pregnancy outcomes were compared accord-
ing to C levels. Results: The prevalence of pregnancy complications
was higher in women with PAPS (31,7% vs 9,3%; p¼0,01).
NormalC3/C4 LowC3/C4 NormalC3/C4 LowC3/C4
PAPS PAPS Controls Controls
Complications (n¼33) (n¼8) (n¼30) (n¼13)
Spontaneous 2 1
abortions
Fetal death 1 1
Preeclampsia 1 1
IUGR 1 2
PROM 1 1
Oligoidramnios 2
Increase uterine 2 1
artery resistance
Lupus
Hypocomplementemia was found in 19,5% of APS pregnancy and in
30,2% of controls. Among cases, the complication prevalence was
higher in women with hypocomplementemia (62,5%) in comparison
with women with normal C (24,2%)(p¼0,03). In the control group
there was no significant difference in the prevalence of pregnancy com-
plications between women with hypocomplementemia (7,7%) and
women with normal C (10%) (p¼1). Conclusion: Complement con-
sumption was significantly associated with pregnancy complications
in PAPS. No significant correlation was observed between C level
and pregnancy outcome in the control group. Our data suggest that
hypocomplementemia might be a negative prognostic factor in PAPS
pregnancies, supporting the experimental evidence that C is a mediator
of aPL damage to the developing fetal–placental unit.
Disclosure: Nothing to disclose.
B066
C’ involvement in aPL-mediated placental damage: prospective study in
APS pregnant women
Meroni PL1, Tedesco F2, Tripodo C3, Borghi MO1
1
University Milan, Milan, Italy; 2University Trieste, Trieste, Italy;
3
University Palermo, Palermo, Italy
Background: Recurrent fetal losses are associated with pathogenic aPL.
Still debated are the mechanisms involved. C’ activation was reported
to play a key role in murine models of aPL-induced fetal loss but still
debated is whether comparable findings can be found in pregnant APS
patients. Aims: to investigate C’ deposition in placental tissues from
PAPS pregnant women in a prospective study and its relationship with
outcome, histological lesions & therapy. Methods: PAPS were classi-
fied according to the Sydney criteria. Placentae at delivery or after
abortion were collected together with clinical & laboratory data.
C’ deposition was evaluated by immunohistochemistry. Results: 14
PAPS patients and 5 normal pregnancies were enrolled; 17 pregnancies
(1 twin pregnancy) in the PAPS and 5 in the control group. 11 placen-
tas after term pregnancy & 6 abortive specimens (5 at 20th & 1 at 25th
wg) were collected in the PAPS group. LMWH & low dose ASA were
used in 15/17 pregnancies, low dose ASA only in 2/17 as first pregnan-
cies, IVIg and low dose corticosteroids were also used in 1 patient
because of no response to LMWH & ASA. Decidual vasculopathy
was detectable mainly in PAPS. Intervillous thrombi were found in
PAPS cases, while villous infarcts and foci of villitis were detected
also in controls. C’ deposits were constantly detected in PAPS while
absent in controls. C1q & C3 were both detected on the endothelium of
decidual vessels showing signs of IgG/IgM deposition, as well as in the
interstitium among decidual stromal cells at sites of inflammation.,C9/
TCC showed a subendothelial distribution in decidual vessels.
Similarly to Igs, C1q, C3 and C9/TCC were also detected on the sur-
face of the villous syncitiotrophoblast, with C9/TCC deposits being
more conspicuous in infarcted villi. Conclusions: i) histological analysis
does not show any specific pattern and no signs of widespread inflam-
mation in agreement with previous reports; ii) C’ activation was
reported for the first time in human APS placentas obtained in a pro-
spective study; iii) C’ deposition was found both in abortive
specimens and in placentas at term, and there was no relationship
with therapy.
Disclosure: Nothing to disclose.
 Abstracts

517
B067 Conclusions: Patients with APS plus another thrombophilia were full
anticoagulated from the beginning of the pregnancy and had the low
A longitudinal study of antiphospholipid antibodies in women at risk of rates of placental abnormalities. Both APS and APS LT had similar
preeclampsia rates of placental abnormalities.
Bertolaccini ML1, Murru V1, Cuckson C2, Caleyachetty R1, Seed P2,
Disclosure: Nothing to disclose.
Poston L2, Khamashta MA1
1
The Rayne Instituite, King’s College London School f Medicine,
London, United Kingdom; 2Maternal and Fetal Research Unit, King’s
College London School of Medicine, London, United Kingdom B069

Purpose: Preeclampsia in APS pregnancies has been reported in 32% Fetal inherited thrombophilia and malformations of vascular origin:
to 50% of the cases. However, the prevalence and clinical importance preliminary data
of aPL in the general obstetric population presenting with preeclamp- Couto E1, Zaccaria R1
1
sia remains uncertain. We evaluated the prevalence and behaviour of Unicamp, Campinas, Sao Paulo, Brazil
aPL in a cohort of pregnant women in whom preeclampsia developed
during follow up and compare the results with a group of women who The objective was to evaluate the association between fetal inherited
did not develop this condition. Patients and Methods: 79 women were thrombophilia and malformations of vascular origin. The mutations
divided into 2 groups: 1) high risk women (HRW), identified by a Factor V Leiden, G20210A in FII gene and C677T in the methylene
previous history of preeclampsia (n¼45, mean age 31.96) and 2) tetrahydrofolate reductase gene were investigated in two groups of
low risk women (LRW) comprising those who attended the hospital fetuses. In the first group there were 50 fetuses with abdominal wall
antenatal clinics (n¼34, mean age 31.46). IgG/IgM aCL and anti- or neural tube defects, porencephaly, hydranencephaly or limb reduc-
b2GPI and LA were measured over gestation. Results: aPL were pre- tion defects, whose mothers were outclinic patients at the Fetal
sent in 6/79 women (3/45 HRW and 3/34 LRW). Fifteen women devel- Medicine Prenatal Clinic from UNICAMP - Campinas University.
oped preeclampsia, all from the HRW group. 2/15 patients with The DNA for the detection of thrombophilic mutations was obtained
preeclampsia had persistently positive aPL (1 IgG aCL and anti- from routine cordocentesis for fetal cariotyping, realized from 2001 to
b2GPI and 1 LA). Prevalence of aPL was not different between 2005. In the second group, 100 morphologically normal newborns
patients who did and those who did not develope preeclampsia (2/15 delivered at UNICAMP - Campinas University Maternity Hospital
vs. 4/64, OR 2.3 [95%CI 0.4-14], p¼0.3). IgG aCL and anti-b2GPI were included. The blood samples for detection of thrombophilic
mutations were obtained immediately after placental detachment.
remained positive all through pregnancy. LA became negative in most
The mutation G20210A in FII gene was not detected in any of the
of the positive cases (3/4) during follow up. Conclusion: The presence
studied blood samples. No significant difference was encountered in the
of aPL does not predict the development of preeclampsia in the general
detection of C677T mutation between both groups (8% in Group 1
obstetric population.
and 7.3% in Group 2). The Factor V Leiden was significantly more
Disclosure: Maria Laura Bertolaccini is funded by the Louise Gergel common in Group 1 (14%) than in Group 2 (3.1%). In conclusion,
Fellowship. fetal thrombophilia, in special Factor V Leiden, may play a role in fetal
malformations of vascular origin. More studies in this direction are
necessary to confirm this hypothesis.
B068
Disclosure: Nothing to disclose.
Placental histological abnormalities in patients with Antiphospholipid
Syndrome (APS)
Barros V1, Schultz R1, Baptista F1, Bortolotto MR1, Oliveira A1, B070
Zugaib M1
1
Clinics Hospital University of Sao Paulo, Sao Paulo, Brazil Late Obstetric Complications in Antiphospholipid Syndrome: Clinical
Presentations and Management
The histological abnormalities of the APS placentas reflect the success Cohen H1, Hills J1, O’Brien P1
of the anticoagulation in this thombophilic pathology. We have retro- 1
University College London Hospitals, London, United Kingdom
spectively studied 27 placentas. The APS placentas were studied in
three groups: APS (Sapporo’s criteria), APS low titer (APSLT, antic- Late obstetric complications are recognised to occur in antiphospholi-
ardiolipin antibodies < 40) and APS with a combined thrombophilia pid syndrome (APS) although data supporting diagnostic criteria and
(CT). The treatment employed was enoxaparin in 98% of patients. management remain limited. The Sydney criteria for late obstetric
Patients with APSLT and no previous thrombosis received 40mg/day morbidity in APS include one or more preterm births of a morpholo-
enoxaparin, APS enoxaparin 1mg/kg/day (recent thrombosis 2mg/kg/ gically normal neonate at 34 weeks of gestation because of severe pre-
day) and APSCT (2mg/kg/day). eclampsia or eclampsia or severe placental insufficiency. We analysed
107 pregnancies in 30 women who had a history of late obstetric com-
plications associated with persistent antiphospholipid antibodies
TYPE OF LESION APS APS LT APS CT TOTAL
(aPL). aPL positivity was based on testing for lupus anticoagulant
Number of placentas/number 13/47 12/85 2/25 27/157 (LA) according to ISTH guidelines and IgG and IgM aCL and
pregnancies studied ab2GP1 by standardised ELISA. In 64 untreated pregnancies, out-
infarction 3(6.3%) 5(5.8%) 0 8(5%) comes were: live births 13 (23%); pregnancy losses >20 weeks/perina-
massive intervilous fibrin 3(6.3%) 3(3.5%) 0 6(3.8%) tal mortality 16 (25%), intrauterine growth restriction (IUGR) 9
deposition (14%), pre-eclampsia 7 (11%), placental abruption 3 (5%), HELLP
other thrombotic lesions 3(6.3%) 2(2.3%) 1(4%) 5(3.18%) 2 (3%) and 35 (55%) pregnancy losses <20 (18 <10 and 17 between 10-
acute deciduitis 2(4.2%) 1(1.2%) 0 3(1.9%) 20) weeks (some women had more than one manifestation). The mean
retroplacental hematoma 2(4.2%) 1(1.2%) 1(4%) 3(1.9%) gestation of live births was 34 (range 21–40) weeks and 7/17 (41%)
babies were delivered at 34 weeks gestation. 7/30 (23%) women also

Lupus
 Abstracts
518
had a thrombotic history (5 venous thromboembolism and 2 a transi-
ent ischaemic episode (TIA)). 20 of the 30 women (1 with a history of a
TIA) had 43 pregnancies treated in our unit with prophylactic dose low
molecular weight heparin (LMWH) and low dose aspirin (LDA) 75 mg
daily. The outcomes of these index pregnancies were: live births 31
(72%); pregnancy losses >20 weeks/perinatal mortality 2 (5%),
IUGR 1 (2%), pre-eclampsia 2 (5%), placental abruption 3 (7%)
and (8 (19%)) pregnancy losses <20 (mean 10; range 5-19) weeks.
The mean gestation of live births post-treatment was 39 (range 28–
40) weeks and 1/31 (3%) babies were delivered at 34 weeks gestation.
Our data suggest that Sydney clinical criteria for late obstetric APS
appear generally applicable, however, a proportion of patients would
be excluded. A live birth was achieved in >70% with prophylactic
LMWH and LDA; however, some women may benefit from more
intensive regimens.
Disclosure: Nothing to disclose.
B071
Diagnostic Criteria for Obstetrical Antiphospholipid Syndrome (OAPS).
Results of a questionnaire of the European Forum on Antiphospholipid
Antibodies
Boffa MC1, Cohen H2, Tincani A3
1 pair’, ‘confused’, ‘heart wrenching’, ‘panic’, and ‘devastated to describe
Hopital Jean Verdier, AP Hopitaus de Paris, Bondy, France;
2 their reactions to their losses and the unexpected diagnosis.
University College London Hospitals, London, United Kingdom;
3 Delayed Diagnosis: The women in this study reported the challen-
Spedali Civili di Brescia, Brescia, Italy
Recently OAPS prognosis has been improved to investigate which
patients we are treating and on the criteria we are selecting them.
OAPS appears to represent a specific entity which differs markedly
from thrombotic APS. It is a clinically heterogeneous syndrome.
Early pregnancy losses should be distinguished from late obstetric
complications as well as purely obstetric forms from those associated
with thrombosis or autoimmune diseases. We suspected a dissociation
between the clinical and laboratory diagnosis of OAPS based on the
Sydney criteria and the criteria used by experts to underpin therapeutic
decisions in clinical practice. We therefore, prior to the Marseilles
meeting of the European Forum, sent a questionnaire to the 18
APL-expert participating teams to enquire about which symptoms
and laboratory data they favour to treat patients. We summarise
here responses to the part of the questionnaire that focused on scenar-
ios where APS diagnostic criteria did not fulfil Sydney criteria. To the
first question: are the Sydney criteria appropriate to define OAPS 14/18
answered ‘no’; to the following questions: do you consider and treat as
OAPS a pregnant woman: A) with Sydney obstetric criteria and i) low
titre APL: 17/18 answered ‘yes’ (11 with both low dose aspirin (LDA)
and LMW heparin); and ii) previously APL positive, and negative at the
time of pregnancy: 15/18 answered ‘yes’; B) with Sydney laboratory
criteria and i) IUGR at term (and not with prematurity) 17/18 ‘yes’;
ii) with HELLP syndrome 18/18 answered ‘yes’; iii) two early miscar-
riages 15/18 answered ‘yes’. Finally, experts were asked do you treat
during pregnancy aPL-positive women with no clinical symptoms: 17/18
stated that they would treat, 13 with LDA alone, 4 with both LDA and
LMW heparin. On the basis of the experts’ answers, it appears that the
Revised Classification criteria are not so useful to identify OAPS
patients for therapeutic purposes. The results suggest that, for
OAPS, the specific classification criteria should be reviewed or new
diagnostic criteria be established.
Disclosure: Nothing is disclosed.
Lupus
B072
Explaining ‘‘Unexplained’’ Loss: Women’s Experiences of Perinatal
Losses with Antiphospholipid Syndrome
Mathew S1
1
University of Texas medical Branch, Galveston, Texas
Purpose: To explore women’s experiences of pregnancy losses with
APS. Methods: The participants included thirty-eight women from 5
countries, who were members of perinatal loss online support groups,
and had 1 or more perinatal losses with APS. The eligibility criteria for
participation in the study included the presence of a medically diag-
nosed primary or secondary APS and the experience of1 or more
pregnancy losses during the past 15 years. Women who felt they met
the eligibility criteria were invited for semistructured email interviews.
Summary of the Results: The analysis of the data yielded two over-
arching themes: Existence in Bewilderment and Persistence in the
Quest for Knowledge and Information. The first major theme has
two sub themes: Delayed Diagnosis and Living in Uncertainty.
Existence in Bewilderment: Women in this study described the
entire experience of perinatal losses, before and after diagnosis of
APS, as having to exist in a somewhat confused state of mind, mixed
with a perplexity of psychosocial emotions. The emotions of losing
their babies and learning that those losses were caused by their own
blood disorder were inseparable. In general, they used the terms ‘des-
ging experiences of getting diagnosed with APS and having the mystery
of their unexplained losses revealed.
Living in Uncertainty: Women in this study reported the feeling of
living in uncertainty in regards to perinatal outcome and other APS
complications, even after the diagnosis. From their reports, several
physicians seemed not to be knowledgeable about the extent of
damage APS could cause in women’s lives.
Persistence in the Quest for Knowledge and Information: The parti-
cipants in this study had the curiosity to learn about APS using dif-
ferent sources of information to get a better understanding of this
condition. Seeking information became a necessity rather than a
desire. Conclusion: The themes and the subthemes are interrelated
and give a comprehensive picture of what it is like living with APS
during and beyond pregnancy. Delayed diagnosis of APS, treatment
failure, and challenges of living in uncertainty led the women to an
existence in bewilderment and persistence in the quest for knowledge.
Disclosure: Nothing to disclose.
B073
Antiphospholipid syndrome and pregnancy: comparative results with non
A.P.S population, Buenos Aires, Argentina
Latino JO1, Pechini S2, Fernandez Romero D1, Huespe M2,
Casavilla JJ1, Luca R2
1
Durand Hospital, Buenos Aires, Argentina; 2Santojanni Hospital,
Buenos Aires, Capital Federal, Argentina
Introduction: The A.P.S is the most frequently acquired thrombophilia
and is linked to adverse gestational events such as recurrent abortion,
prematurity and intrauterine death. Purpose: Evaluate the efficacy of
the implementation of a protocol for the clinical, obstetric and
therapeutic care of patients with A.P.S during the gravid puerperal
state regarding the ABORTION, PREMATURITY and

INTRAUTERINE DEATH variables, comparing them with a NON
A.P.S CONTROL GROUP. Material and Methods: Design:
Observational prospective cohort study. Population: Ø A.P.S Group:
138 patients, Ø Non A.P.S Group: 155 pregnant women. Care and
Follow-up Protocol: 1) Monthly general and immunologic laboratory,
2) Strict fetus checkup: monthly ecography, doppler ecography from
week 24th on, fortnightly, and NST from week 28th on (one a week)
and 2 NST/week, from week 32 on. Treatment: It consists of the
administration of:ASA _(Acetylsalicylic Acid),
LMWH (Low-
Molecular-Weight Heparin): Enoxaparin, and
Gamma globulin
(g globulin). Considered variables: Abortions, Prematurity, and
Intrauterine Death. Results: A.P.S patients treated: Abortions 10%,
Prematurity 17.7%, Intrauterine Death 6.5%. Control Group:
Abortions 14.8%, Prematurity 4.5% and Intrauterine Death 3%.
Conclusions: a) The abortions rate in the treated patients group
(10,1%) and in the control group (14.8%) ¼ (p¼0.23) is similar. b)
The prematurity rate is higher in the group of patients with A.P.S
(17.7%) than in the control group (4,5%) (p¼0,001). c) The intrauter-
ine death rate in the group of patients with A.P.S (6.5%) and in the
control group (3%) (p¼0.20) is similar. The application of the pro-
posed protocol determines results similar to the NON A.P.S popula-
tion with respect to the ABORTION and INTRAUTERINE DEATH
variables.
Disclosure: No response indicated.
B074
Characterization and study of systemic lupus erythematosus associated
with secondary antiphospholipid syndrome
Rodrı´guez Moreno M1, Cervera R2, Espinosa G2, Miranda Leon MT3,
Guzmán Úbeda MA1, Rodrı´guez Moreno I4
1
Virgen de las Nieves University Hospital, Granada, Spain;
2
Hospital Clı´nic, Barcelona, Spain; 3University of Granada, Granada,
Spain; 4Granada, Spain
Design: See title. Methods: 44 patients with SLE-APS.
– Variables: See results
– Diagnostics criteria: Revised criteria: ACR SLE 1999, APS cri-
teria 2006.
– Statistical Method: SPSS 15.0
Results: Number of APS criteria were correlated with anticardiolipin
antibodies (aCL) (P¼0.005). Subjects with 2 APS criteria, 66.7% have
negative aCL. Patients with 3 or 4 APS criteria, 100% had positive
aCL. Number of SLE criteria plus APS criteria were correlated nega-
tively with Complement C3 values(P¼0.01). Value of DNA antibody
was correlated negatively with BUN value (P¼0.0017). In patients with
SLE-SAPS: Complement C3 and C4 values were correlated
positively(rS¼ 0.502; P¼0.015). Renal function parameters BUN and
ANA were negatively correlated (rS¼ -0.580; P¼0.009). And finally,
kidney function parameters, BUN and Creatinine were correlated
positively(rS¼ 0.803; P¼0.000012). Conclusion: 1- Patients with SLE-
SAPS that have  3 APS criteria have positive aCL in 100% of the
cases.(OR¼ 63; 95%C.I.(2.55; 1558.4). 2- In patients with SLE-APS,
Complement C3 values are correlated negatively with number of SLE
criteria and also with number of SLE plus APS criteria. 3- DNA
autoantibodies were correlated positively with ANA and negatively
with BUN values, in patients with SLE-SAPS. 4- Complement C3
and C4 values in patients with SLE-SAPS were correlated positively.
5- Renal function parameters BUN and ANA were negatively corre-
lated in patients with SLE-SAPS. 6- And finally, in patients with
Abstracts
519
SLE-SAPS, renal function parameters BUN and Creatinine were cor-
related positively.
Disclosure: Nothing to disclose.
B075
Comparative study between hypergammaglobulinaemia and monoclonal
gammopathy of undetermined significance in presence of systemic lupus
erythematosus associated with secondary antiphospholipid syndrome
Rodrı´guez Moreno MDR1, Cervera R2, Espinosa G2, Miranda MT3,
Guzman MA1, Rodriguez Moreno I4
1
Virgen de las Nieves University Hospital, Granada, Spain;
2
Hospital Clı´nic, Barcelona; 3University of Granada, Granada, Spain;
4
Granada, Spain
Design: See Title. Methods: 44 patients SLE-APS (normal, hypergam-
maglobulinaemia and MGUS).
– Variables: See results
– Diagnostics criteria: Revised criteria:ACR SLE (1999), APS 2006
– Statistical Method: SPSS 15.0.
Results: Prevalence MGUS in SLE-APS 9% with 95% C.I (1.52%;
29.51%), MGUS type Ig G Kappa was the most frequent. Age
in patients with SLE-APS and MGUS 61.67þ/-17.78; SLE-APS
and Hypergammaglobulinaemia 53þ/-12.18; SLE-APS without
Hypergammaglobulinaemia or MGUS 39.56þ/-11.42 (p¼0.022). In
patients with SLE-APS and MGUS: Rheumatoid Factor (RF) have
a higher value (p¼0.004). DNA antibody are lesser (p¼0.08). Anti-Ro/
SSA 100% (p¼0.11). Anti-La/SSB 66.7% (p¼0.039). Anticardiolipin
antibody of Ig M are related with MGUS (p¼0.106). In patients with
SLE-APS and Hypergammaglobulinaemia we have more patients
treated with immunosuppressant drugs (p¼0.087). The only 3 cases
of death are happened in patients with SLE-APS and Hypergamma-
globulinaemia, and it not exist any case in the others. If we bracket in 2
categories (Hypergammaglobulinaemia opposite MGUS plus normal)
(p¼0.093) Conclusion: 1- Patients with SLE-APS and MGUS have
higher age than without MGUS or with Hypergammaglobulinaemia.
2- Acute phase reactants RF are increased in all patients with SLE-
APS and MGUS and have higher values than in the others. And we
haven’t observed differences in terms of the ESR and CRP. 3- Mean
values of DNA antibody are lesser in patients with SLE-APS and
MGUS respecting the others and this result is near to the significance.
4- Anti-Ro/SSA and Anti-La/SSB autoantibodies was present in 100%
and 66.7% of the patients with SLE-APS and MGUS respectively.
Anti-La/SSB was more frequent in patients with SLE-APS and
MGUS than in the others. 5- Hyper-gammaglobulinaemia in patients
with SLE-APS may be a risk factor of mortality. 6- It’s necessary to
follow the evolution of MGUS each 6 months.
Disclosure: Nothing to disclose.
B076
Risk factors to affect the mortality and morbidity in patients with
antiphospholipid syndrome: a longitudinal follow-up study of 95 Japanese
patients
Fujieda Y1, Atsumi T1, Otomo K1, Kato M1, Amengual O1, Yasuda S1,
Horita T1, Koike T1
1
Hokkaido University Graduate School of Medicine, Sapporo,
Hokkaido, Japan
Objective: To clarify the risk factors to affect the morbidity and
mortality in patients with antiphospholipid syndrome (APS).
Lupus
 Abstracts
520
Patients and Methods: This study comprised 95 patients with APS
(female 81, mean age 45). The mortality rate, recurrence of thrombotic
events, and bleeding episodes during the follow-up period were com-
pared with the clinical profiles of the patients. Kaplan-Meier method
was introduced to define risk factors for recurrent thrombosis and
mortality. Results: The mean follow up period was 8 years (2-21).
Thrombotic events recurred in 27 (28%) patients and recurrence rate
was 4.1 per 100 patient-years. The mean time between the initial
thrombosis and the first recurrence was 5.5 years (0.3-18). The arterial
events recurred in 20 patients and venous events in 7. A total of 5 (5%)
had serious bleeding events and 9 (10%) patients died during the
follow-up period. In Kaplan-Meier analysis, Diabetes Mellitus (DM)
had significantly affected the recurrence of thrombosis and mortality
(p¼0.01), but other factors investigated (antibody profile, age, gender,
complication of SLE) did not correlated with the prognosis.
Conclusions: Patients with APS developed frequent recurrent throm-
bosis with high mortality despite current anticoagulant treatment. DM
is a risk for recurrent thrombosis and mortality in APS patients.
Disclosure: Nothing to disclose.
B077
Characterization and study of systemic lupus erythematosus associated
with secondary antiphospholipid syndrome and monoclonal gammopathy
of undetermined significance
Rodrı´guez Moreno MDR1, Cervera R2, Espinosa G2, Miranda
Leon MT3, Guzmán Úbeda MA1, Rodrı´guez Moreno I4
1 phospholipid profiles. Patients with antiphospholipid syndrome have
Virgen de las Nieves University Hospital, Granada, Spain;
2 increased arterial stiffness, in particular patients with arterial throm-
Hospital Clı´nic, Barcelona, Spain; 3University of Granada, Granada,
Spain; 4Granada, Spain
Design: See title. Methods: 44 patients with SLE-APS.
– Variables: See results
– Diagnostics criteria: Revised ACR criteria for SLE (1999),
Preliminary classification criteria APS of the international work-
shop (1999).
– Statistical Method: SPSS 15.0
Results: In patients with SLE-APS: Prevalence of MGUS was 9% with
95% C.I (1.52%; 29.51%) and MGUS type IgG Kappa was the most
frequent. Age in patients with SLE-APS and MGUS had a meanþ/-
SEM of 61.67þ/-17.78 and without MGUS of 46.95þ/-13.42(p¼0.05).
In patients with SLE-APS and MGUS Rheumatoid Factor (RF) had a
higher value than in the others (p<0.005). Anti-Ro/SSA was present in
100% of the cases (p¼0.063). Anti-La/SSB was present in 66.7% of the
cases (p¼0.041). Anticardiolipin antibody of IgM isotype in serum of
the patients with SLE-APS was related with the presence of MGUS
(p¼0.10). Conclusion: 1- The prevalence of MGUS in patients with
SLE-APS was 9% with 95% C.I (1.52%; 29.51%) higher which in
general population or SLE; and MGUS type IgG Kappa was the
most frequent observed in our study. 2- The patients with SLE-APS
and MGUS had higher age than without MGUS. Differences in
terms of the sex, smoking, number of SLE criteria or number of
APS criteria were not observed. 3- Acute phase reactants RF were
increased in all patients with SLE-APS and MGUS and had higher
values than in the others. Differences in terms of the ESR and CRP
were not observed. 4- Anti-Ro/SSA and Anti-La/SSB autoantibodies
were present in 100% and 66.7% of the patients with SLE-APS and
MGUS respectively. 5- It’s necessary to follow the evolution of MGUS
each 6 months.
Disclosure: Nothing to disclose.
Lupus
B078
Vascular stiffness is increased in antiphospholipid syndrome: significance
and clinical associations
Wahl D1,2,3, Salvi P1, K-Schwartz A2, Bravetti V2, Moline T2,
Farese E2, Regnault V3, Lacolley P3, Valla M2, Zuily S1, Guillaume C2,
Lecompte T1, Benetos A1
1
Nancy University, Vandoeuvre, Lorraine, France; 2CHU, Vandoeuvre,
Lorraine, France; 3INSERM U961, Vandoeuvre Les Nancy, Lorraine,
France
Vascular stiffness is an independent risk factor of cardiovascular events
in the general population. In order to determine whether arterial stiff-
ness was increased in patients with antiphospholipid antibodies we
studied 53 patients with antiphospholipid antibodies and 109 controls
matched for age and sex. Patients were: 46.02.3 years-old, 40 female
and 13 male. Twenty-six had primary antiphospholipid syndrome,
16 antiphospholipid syndrome associated with SLE. Clinical manifes-
tations included arterial thromboembolic events n¼14, venous throm-
boembolism n¼28, obstetrical manifestations of APS n¼9 and
11 patients were asymptomatic. Pulse wave velocity (PWV) was used
to investigate arterial stiffness and was determined by aplanation tono-
metry. PWV was increased in patients: 8.14  0.40 m/s and 7.24  0.20
m/s in controls (p¼0.036). PWV was correlated with age
(r¼0.75;p¼0.0001) and systolic blood pressure (r¼0.62;p¼0.0001).
PWV was significantly increased in patients with arterial thrombosis
than in controls (9.541.20 m/s and 7.230.20 m/s, respectively;
p¼0.0021) or patients without arterial thrombosis (7.630.40 m/s;
p¼0.0416). PWV was not significantly different among various anti-
botic events.
Disclosure: Nothing to disclose.
B079
Cardiovascular Disease (CVD) Prevention Counseling Program for
Lupus and/or Antiphospholipid Antibody (aPL) Positive Patients
Haiduc VF1, Richey M1, Tzakas S1, Konstantellis L1, Erkan D1
1
Hospital for Special Surgery, New York, NY
Purpose: To determine the satisfaction of lupus and/or aPL-positive
patients who participate in a counseling program that has been devel-
oped to increase the awareness of CVD risk factors as well as throm-
bosis prevention strategies. Methods: A free-of-charge counseling
program, which is partially supported by NY Community Trust, has
been developed to provide a basic assessment of and education about
the CVD and thrombosis risk factors in lupus and/or aPL-positive
patients. The assessment phase includes the evaluation of blood pres-
sure and glucose, cholesterol profile, waist circumference, BMI, diet,
exercise, and smoking habits, family history of CVD, homocysteine
and hs-CRP levels, Framingham CVD risk, aPL-profile, and medica-
tions. The education phase includes discussion of the above risks as well
as CVD and thrombosis prevention strategies. At the end of the coun-
seling, patients: a) receive tailored lifestyle recommendations; b) are
referred to the nutrition and physical therapy departments as needed;
and c) complete an anonymous survey to evaluate the program.
Patients are followed every 3–6 months. We report the results of
patients’ initial visit satisfaction surveys (Likert Scale; 1: best to 5:
worst). Results: Since the launch of the program in March 2009, 64
patients have received counseling and 55/64 (86%) completed the
survey. 45% of the participants were not aware of their CVD risk.
 Abstracts

521
The quality of the counseling was reported as (mean  SD) 1.2  0.4; Disclosure: Nothing to disclose.
quality of the educational materials 1.3  0.7; likelihood of recom-
mending the program to others 1.3  0.6; improvement in patient’s
knowledge about CVD risk factors 1.7  0.8; likelihood of the pro-
gram improving patients’ diet 1.8  1; and likelihood of the program B081
improving patients’ exercise pattern 2.1  1.2. Conclusion: Lupus and/
or aPL-positive patients are well satisfied with a free CVD prevention A comparison of the prevalence of antiphospholipid antibody subtypes in
counseling program. The three-year longitudinal analysis of program lupus and in stroke
patients will determine the true effectiveness of the program with Broder A1, Putterman C1
1
respect to decreasing the prevalence of cardiovascular disease risk Albert Einstein College of Medicine, Bronx, NY
factors.
Background: The prevalence of antiphospholipid antibody (aPL Ab)
Disclosure: Nothing to disclose. subtypes in different diseases has not been well studied. We compared
the prevalence of aCL, aPS, and anti-b2GPI IgA, IgG, and IgM anti-
bodies in SLE and CVA. Methods: We included all patients with ICD9
B080 for SLE or CVA diagnosed between 2006 and 2009. Patients with both
diagnoses were excluded. APL Ab positivity (aPL Abþ) was defined as
A Cross-Sectional Analysis of Patients Referred for Evaluation of APS  40 GPL. Results: 217/1136 (19%) of SLE and 97/5827 (1.6%) of
Barbhaiya M2, Keles F3, Erkan D1,2, Lockshin M1,2 CVA patients had all 9 aPL subtypes measured. 185 SLE and 79 CVA
1
Hospital For Special Surgery, New York, NY; 2Cornell Medical patients were included. Compared with the CVA group, the SLE group
College, New York, NY; 3Hacettepe University, Ankara, Turkey was younger and included more women. There was no difference in the
time to aPL measurement between the 2 groups (257 days IQR (59,
Background: The Updated Sapporo Criteria (USC) provide a uniform
591) vs. 279 days IQR(15, 727). The prevalence of different aPL Abþ
approach for research and are also important in practice, in which
subtypes, shown in Table 1, was between 0% and 8.7%. Only the
APS ‘‘over-diagnosis’’ is not uncommon. Purpose: To analyze the
prevalence of anti-b2GPI IgGþ was statistically significantly higher
aPL and clinical characteristics of patients referred to our center
in SLE vs. CVA (7.6% vs. 1.3%, P¼0.045).
within the last 5 years with at least one aPL result and evaluation of
APS. Methods: We retrospectively recorded demographics as well as
aPL tests and criteria/non-criteria (NC) manifestations based on USC.
Table 1 The prevalence of various aPLþ subtypes in SLE and CVA
aCL/ab2GPI were grouped as: negative (0-19U); low-titer (20-39U);
and moderate-high titer (>40U). ‘‘Significant aPL’’ was defined as a N(%) aPL Abþ SLE 185 CVA 79 p-value*
positive LA test and/or moderate-high titer IgG/M aCL or ab2GPI
tested twice  12w apart. Results: 45 patients (female 87%, mean aPS IgA 0 (0) 1(1.3) 0.3
age: 41.312.5; and other autoimmune diseases: 20%) were analyzed. aPS IgG 4 (2.2) 1(1.3) >0.999
At the time of referral: 16 (36%) had vascular events (VE); 9 (20%) aPS IgM 4 (2.2) 3 (3.8) 0.4
pregnancy morbidity (PM); 5 (11%) VE & PM; 9 (20%) NC features aCL IgA 3 (1.6) 2 (2.5) 0.6
only; and 6 (13%) aPL only. Based on the referral aPL, 27/45 (60%) aCL IgG 10 (5.4) 4 (5.1) >0.999
had at least one-time positive LA and/or aCL/ab2GPI > 20U; however aCL IgM 3 (1.6) 3 (3.8) 0.37
only 11/45 (24%) of the referrals were based on at least two aPL tests Anti-b2GPI IgA 16 (8.7) 2 (2.6) 0.11
at two different time points. Based on final diagnostic assessment, 21/45 Anti-b2GPI IgG 14 (7.6) 1 (1.3) 0.045
(47%) had ‘‘significant aPL profiles’’ and 8/45 (18%) fulfilled USC Anti-b2GPI IgM 5 (2.7) 4 (5.1) 0.46
(Table). *Fisher’s exact test.

Final Assessment N: 45 The prevalence of at least one aPL Abþ was 29/185 (16%) in SLE
group, and 7/79 (9%) in the CVA group. Among patients who had at
– APS; Significant aPL 8 (18%) least one aPL Abþ, the prevalence of anti-b2GPI IgGþ and anti-
– Possible APS; Significant aPL (>5y between aPL & 5 (11%) b2GPI IgAþ was higher in SLE vs. CVA: 48.3% vs. 14.3%, p¼0.1;
event)
55.2% vs. 28.6%, p¼0.2 respectively. The prevalence of aCL IgM and
– Possible APS; Significant aPL (LA [þ] on anticoagu- 3 (7%)
lation & aCL/ab2GPI [-])
aPS IgM was lower in the SLE vs. CVA: 10.3% vs. 42.9%, p¼0.07;
– Asymptomatic aPL or NC features only; Significant 5 (11%) 13.8% vs. 42.9%, p¼0.12, respectively. We found no statistically sig-
aPL nificant associations between aPL Abþ and age, gender, ethnicity or
– Possible APS; Low-titer aCL/ab2GPI & LA (-) 3 (7%) season in bivariate or multivariate analysis. The total number of aPL
– LA/aCL/ab2GPI (-) 21 (47%) Ab positive subtypes was associated with ethnicity. Conclusion: The
prevalence of clinically significant aPL Abþ is very low even in this
highly selected subgroup of SLE and CVA patients. Different aPL Ab
Conclusion: 47% of patients referred for evaluation of APS have nega- subtypes may be associated with these diseases. Several clinically sig-
tive LA test and aCL/ab2GPI (<20U) based on their final diagnostic nificant differences did not reach statistical significance because of the
assessment. Although manifestations in the remaining of patients may relatively small sample size.
be attributable to aPL, only one-fifth truly fulfill the Updated Sapporo
Criteria. Our results have implications for both physician training and Disclosure: Nothing to disclose.
research.

Lupus
 Abstracts
522
B082
Hematologic Manifestations Among Patients Who Fulfill the Sidney
Laboratory Criteria for APS: a 6 Year Follow-up
Comellas-Kirkerup L1, Molina-Hernández G1, Cabral AR1
1
INCMNSZ, Mexico City, DF, Mexico
Background: To study patients with thrombocytopenia (TP), hemolytic
anemia (HA) or both who meet the APS Sidney laboratory criteria but
do or do not fulfill the clinical criteria. Methods: We reviewed the
medical charts of 187 patients with primary APS from our Registry
(Jan 1986- Dec 2008) and selected those with: a) one or more Sidney
laboratory criteria, and, b) thrombocytopenia (PLTs<100,000 ml) or
HA (Coombsþ, corrected reticulocyte count >2%, elevated indirect
billirrubin and LDH) or Evans’syndrome (ES). Patients with SLE and/
or positive anti-dsDNA (ELISA) were excluded. Age at onset, follow-
up until last medical appointment (LMA), treatment and clinical APS
status were assessed. We defined response to treatment (RT) as PLTs
100,000 ml at LMA if basal PLTs were >50,000 ml, or PLTs >50,000
ml at LMA if basal PLTs were 50,000 ml; for HA: hemoglobin >10
mg/dl without hemolysis. Relapse was considered when RT (any point
at follow-up) could not be maintained. We used X2, Student t test or
U-Mann Whitney test. Results: 55 patients (44 women) met the inclu-
sion criteria: mean age at disease onset 40  12 years with a median
follow-up of 6.8 years (0.14-36.9). 35 had TP, 14 HA and 6 ES.
25 patients (45%) also met one Sidney clinical criteria (13 thrombosis,
12 pregnancy morbidity). In 4 patients (16%) the hemocytopenia and
the APS clinical criteria started simultaneously, in 6 (24%) the former
preceded the clinical criteria (3 years (0.5-12)), in 15 patients (60%) the
hemocytopenia started 4.4 years later (0-32). 30 patients did
not develop thrombosis or pregnancy morbidity during follow-up
(16 TP, 8 HA, 1 ES). aCL and anti-b2GPI were equally prevalent
amongst the studied groups. A higher prevalence of LAC (63% vs.
30%, p¼0.04) was found in patients with full APS (73% vs. 33%,
p¼0.01; OR¼5.6, 95% CI 1.5- 20.4). There were no differences in
age, type of hemocytopenia, use of prednisone, immunosuppressives,
aspirin, splenectomy, RT at LMA, relapses, basal PLTs or Hb among
studied groups. Conclusion: Patients with aPL related hemocytopenias
may or may not develop an APS clinical criteria. This suggests that
aPL associated with hemocytopenia appear to be pathogenetically dif-
ferent from those associated with thrombosis.
Disclosure: Nothing to disclose.
B083
Thrombocytopenia (TP) in antiphospholipid syndrome
Reshetnyak T1, Vasilenko I2, Kastrikina I2, Nasonov E1
1
Institute of Reumatology of RAMS, Moscow, 115522, Russian
Federation; 2Russian Gerontological Scientific Clinical Center, Moscow,
Russian Federation
Aim: To study the relationship between TP, antiphospholipid antibo-
dies (aPL), thromboses and morphofunctional peculiarity of platelets
in APS. Material and Methods: There were 286 patients with SLE (190
female, 96 male) and 84 patients with primary APS (59 female and 25
male). We determined the morpho-functional status of living platelet
by express-method of vital computer morphometry using computer
phase-inteference microscope (CPM) ‘‘Cytoscan’’ (Moscow, Russia)
in 34 patients. Results: 161 from 286 (56%) patients with SLE had
aPL and 142 (88%) from them had symptoms of APS. During the
disease TP was registered in 16% (20/125) of patients with SLE with-
out aPL and in 46% (75/161) - with aPL. 44 out of 75 cases with TP
were associated with reliable APS, 15 – with probable APS and 16 –
with doubtful APS. There were no significant demographic
Lupus
characteristics between patients with and without TP. In 32/84
(38%) patients with PAPS TP was registered in past history and
during the examination TP was detected in 12 patients, in 12/38
(38%) cases it was combined with thromboses and fetal loss. In 2
patients with PAPS, who had high-positive level of aPL and TP only
avascular necrosis of bone were developed. In multiple-factor analysis
TP showed to be a prognostically unfavourable symptom (p¼0.002).
TP attended CAPS in 14/33 and 2/10 SLE and PAPS cases respec-
tively. A high degree of heterogeneity in platelet populations was dis-
covered: the cell size variations (increasing of geometrical parameters
by 12-29% and decreasing of optical parameters by 7-15%). In APS
patients there was found a higher level of platelet activating state: 48
and 53% - the resting platelets; 49 and 39% - activated forms; 3 and
8%-degenerated forms, respectively. Conclusion: TP in combination
with aPL and SLE is a prognostically unfavourable symptom. APS
and SLE patients had the high level of platelet activating state.
Disclosure: Nothing to disclose.
B084
Total and Free levels of Protein S (PS), PS deficiency, PS activity;
anti-PS antibodies and events of thrombosis in patients with SLE
Scali J1, Pacheco G1
1
Durand Hospital, Buenos Aires, Argentina
Background: Thrombosis occurs in SLE leading to serious morbidity/
mortality. It is crucial to predict which subsets of pts are likely to
develop it. Clotting is regulated by natural anticoagulants:
Antithrombins; Protein C/S; Plasmin/ Fibrinolysis. aPLs abs are present
in 40 to 65 % of SLE. Some protein subsets have been implicated as
targets of aPLs (prothrombin, annexin V, b2 GPI; Protein C and S).
SLE may lead to acquired PS deficiency due to the presence of anti-PS
antibodies, but other aPLs abs could contribute altering protein S
activity (pro-thrombotic state). We study the relation among aPLs
atbs, anti-PS atbs, and thrombotic events in SLE. Study: Cohort of
80 pts (50 SLE and 30 controls), in a three year open, prospective
study.50 SLE adult patients were included (Group I) diagnosed
according to ACR criteria; age ranges between 18-52 years; average
duration of SLE 1.5 to 15 years. Control group (Group II) 30 healthy
women age matched. At entry, no patient was under anticoagulant
treatment (neither Heparin, Acenocumarol nor aspirin or
Clopidogrel). A full clinical history, complete physical examination,
routine and immunological laboratories were performed to all patients;
ANA (IFI), dsDNA (Farr assay), aCLs, anti-B2 GPI antibodies;
Lupus anticoagulant, total Protein S levels, Free Protein S levels,
anti-PS atb (ELISA). Doppler examinations performed in upper,
lower limbs and carotid arteries (for eventual presence of thrombosis).
Results: Total Protein S levels were significantly lower in SLE group,
which also had free protein S deficiency (p< 0.001); in comparison to
Group II. Patients having total PS deficiency, showed SELENA/
SLEDAI indexes higher than patients with normal levels (p< 0.04).
We did not find any relation between PS activity in deficient pts and
disease activity. Conclusions: Independently of the presence of aCLS
(þ) and LA (þ), SLE patients showed deficiency in PS activity, con-
sidered to be related to positivity of anti-PS atb (p< 0.001). The pre-
sence of anti-PS ab strongly correlated with the occurrence of
thrombotic events (55.5% aPS þ vs. 4.7 aPS -). Total Protein S
levels were significantly low in Group I vs. Group II (p< 0.034). aPs
(þ) patients (65%), were also positive for other phospholipids, com-
pared with aPS(-) pts (18%). Protein S activity was significantly low in
patients with positivity to other phospholipids (p< 0.001).
Disclosure: Nothing to disclose.

B085
Antiphospholipid Syndrome in Colombian patients
Tobón GJ1, Osio LF1, Bonilla-Abadı´a F1, Cañas CA1
1
Fundación Valle del Lili, Cali, Valle del Cauca, Colombia
Background: Clinical and immunological data of antiphospholipid syn-
drome (APS) in Latin American patients are limited. Objetive: To
describe the main clinical and immunological manifestations in a
cohort of APS in Colombian patients. Methods: Data were collected
from August 2001 to July 2009. Cases fulfilled Sapporo Criteria for
inclusion. Results: Fifty patients were evaluated. Forty-two patients
were female (84%). Mean-age was 40.1  11.6 years. Mean duration
of disease was 5.5 years. Thirty patients presented a primary APS
(60%). Secondary APS was associated mainly with Systemic Lupus
Erythematosus (SLE) in 18 patients (34%). The most common throm-
botic manifestations were deep vein thrombosis in 26 patients (52%)
and arterial thrombosis in 18 patients (36%), mainly pulmonary
thrombo-embolism (n¼9). The most common pregnancy morbidity
was early pregnancy losses (n¼17 female patients). Livedo reticularis
was the most common cutaneous manifestation (16 patients-32%).
Neurological involvement was present in 21 patients (42%), mainly
migraine (18%), and stroke (18%). Catastrophic APS was present in
two patients. Anticardiolipin IgM antibodies, IgG antibodies and
lupus anticoagulant were positive in 29 patients (58%), 34 (68%)
and 27 (54%), respectively. Conclusion: Few cohorts have reported
the clinical and immunological manifestations in Latin-American
APS patients. There are several differences between Latin-American
and European patients.
Disclosure: Nothing to disclose.
B086
The catastrophic antiphospholipid syndrome - a case report
Furmańczyk A1, Komuda-Leszek E1, Gadomska W2, Windyga J2,
Durlik M1
1
Transplantation Institute, Warsaw, Mazowieckie, Poland; 2Institute of
Hematology and Transfusiology, Warsaw, Mazowieckie, Poland
Background: Catastrophic antiphospholipid syndrome (CAPS) is a
rare disease defined as acute failure of at least three tissues, organs
or systems caused predominantly by microangiopathy confirmed by
histopathology. Description: We present the case of a 39-year-old
male patient with APS of 12 years‘duration with ulceration of the
cruris, treated chronically with heparin. Congenital thrombophilia
and SLE were excluded. A worsening renal function had been observed
since 5 months. On admission the patient required haemodialysis. We
performed a renal biopsy, which revealed thrombotic microangiopa-
thy. A positive ACL titer was obtained >120 GPL (N:<10), 13.71
MPL (N:<7); and anti-b2-GPI >100 GPL (N:<8), 10.69 MPL
(N:<8) and strong LA. According to the criteria introduced by
Asherson, CAPS was diagnosed (skin and renal involvement, small
vessel thrombosis confirmed by histology, positive APLA titers). The
patient had been treated with LMWH, glucocorticosteroids, plasma-
pheresis and IVIG (1g/kg body weight). Renal function improved
(serum creatinine level of 1.5 mg/dl). Discussion: After applied treat-
ment a significant clinical improvement was observed, which was
thought to be associated with the decrease in circulating APLA. In
order to consolidate the clinical outcome patient obtained IVIG. The
next application of IVIG was planned at 3–4 weeks. However, the
patient did not require additional doses of IVIG as no signs of recur-
rent CAPS were observed after 2 months of follow-up. Conclusion: A
promising outcome confirms the necessity and efficacy of applied
Abstracts
523
medications. We suggest that this treatment prevented from developing
multiorgan failure.
Disclosure: Nothing to disclose.
B087
Anticardiolipin antibodies and ischemic stroke due to spontaneous
cerebral artery dissection
Kalashnikova L1, Dobrynina L1, Alexandrova E2, Novikov A2,
Konovalov R1
1
Research Center of Neurology, Moscow, Russian Federation;
2
Institute of Rheumatology, Moscow, Russian Federation
Background: Antiphospholipid syndrome (APS) and spontaneous cer-
ebral artery dissection (D) are two common causes of ischemic stroke
(IS) in young adults. It is considered that weakness of vascular wall
usually predispose to D, rarely it is connected with arteritis. Infection is
believed to be one of the risk factors of D. In some patients D is
associated with anticardiolipin antibodies (aCL). The relationship
between them is not clear. Materials, methods, results: We studied
aCL by ELISA in 60 young patients with IS due to spontaneous cere-
bral artery D. ACL were found only in one patient (1.7%). This was
44-year-old women with D of inner carotid artery (ICA) and middle
cerebral artery. D was verified by MRA which revealed string sign and
by cervical MRI, which showed intramural haematoma in ICA wall.
MRI of the head demonstrated large hemispheric infarct. ACL level at
first study was 88 GPL and 10 MPL, ab2GPI of IgG and IgM isotypes
were 22 and 4 units/ml respectively. ACL elevation was stable (141-95
GPL) for 3 year follow-up period. In one year after first study IgG
ab2GPI increasing (100 units/ml) was found. The patient had neither
peripheral or visceral thrombosis, complicated obstetric history, nor
clinical features of connective tissue weakness. In the course of one
year preceding IS and during follow-up period the patient had mild
arthralgia, mild vertebral pain, anemia, increased blood sedimentation
rate. During follow-up she did not have any new cerebrovascular
events (without oral anticoagulant and aspirin). Conclusion: ACL are
rarely found in patients with spontaneous cerebral artery D. The cause-
and-effect relations between aCL and D appear to be absent. It is
suggested that the same factor (chronic infection?) leads to arterial
wall damage in patients with D and aCL production.
Disclosure: Nothing to disclose.
B088
Echocardiography at diagnosis of antiphospholipid syndrome provides
prognostic information on valvular disease evolution and identifies two
subtypes of patients
Pardos-Gea J1, Ordi-Ros J1, Cortes-Hernández J1, Balada E1
1
Vall Hebrón University Hospital, Barcelona, Barcelona, Spain
Objective: To evaluate whether the presence or absence of valvular
disease at the time of diagnosis of APS predicts its subsequent evolu-
tion. Methods: An initial trans-thoracic echocardiogram was per-
formed on patients at the time of diagnosis of APS. Serial
echocardiograms were conducted along 12 years follow-up. Results:
From 53 patients we found 29(54%) with and 24(45%) without valvu-
lopathy at initial echo. At final echocardiogram, 27 of 29 patients with
initial valvulopathy continued to have valvular disease (93%), and 22
of 24 patients without initial valvulopathy demonstrated absence of
valvular disease (91%). Patients with valvulopathy with respect to
those without, presented more arterial thrombotic events (69% vs
20%, P< 0.001), atherosclerotic risk factors (62% vs. 29%, P¼0.01),
livedo (48% vs 16%, P¼0.01) and migraine (41% vs 12%, P¼0.02).
Lupus
 Abstracts

524
Table 1 Differential characteristics of patients with and without valvulo- hypercoagulable state which is evidenced by the elevated Anti-
pathy at initial echo. b2GP1 antibody level and possible APS.

With Without Disclosure: Nothing to disclose.

valvulopathy, valvulopathy
n(%) n(%) P
B090
Patients 29(100) 24(100)
Primary antiphospholipid syndrome 19(65) 15(62) 0.82 Thrombotic storm: a severe, rapidly progressive thrombotic disorder
Arterial thrombosis 20(69) 5(20) <0.001 Ortel T1, Kitchens C2, Pericak-Vance M3, Erkan D4, James A1,
Venous thrombosis 17(58) 18(75) 0.21 Kulkarni R5, Brandao L6, Hahn S3, Vance J3
1
Pregnancy morbidity 11(38) 7(29) 0.50 Duke University, Durham, NC; 2University of Florida, Gainesville, FL;
3
Risk thrombotic factors University of Miami, Miami, FL; 4Cornell/HSS, New York, NY;
5
Arterial 18(62) 7(29) 0.01 Michigan State University, East Lansing, MI; 6Hospital for Sick
Venous 5(17) 9(37) 0.09 Children, Toronto, ON, Canada
Livedo reticularis 14(48) 4(16) 0.01
Migraine 12(41) 3(12) 0.02 Background: Catastrophic antiphospholipid syndrome (APS), charac-
Raynaud 6(20) 3(12) 0.42 terized by multiple thromboembolic events (TE), occurs in 1% of
Positive IgG anticardiolipin antibodies 24(82) 15 (62) 0.09 patients with APS. A similar rare event, termed ‘‘Thrombotic
Positive lupus-anticoagulant test 28(96) 21(87) 0.21 Storm’’ (TS), occurs in antiphospholipid antibody (aPL) negative
Anticoagulant therapy 22(76) 19(79) 0.77 patients. We hypothesize that patients with CAPS and TS share an
Need for valve surgery 4(13) 0 0.11 underlying genetic risk for this extreme phenotype. Methods: We devel-
Death 7(24) 1(4) 0.06 oped the following criteria independent of aPL status to identify
patients with TS: (1) age<55 yrs; (2) rapid onset of 2 TE (arterial,
a
Comparisons with Chi-square test and unpaired t-test for qualitative and venous or microangiopathic); (3) atypical TE locations; and (4) TE
quantitative variables respectively. P < 0.05 considered significant refractory to standard therapy. Patients with cancer, severe trauma
difference. or recurrent TE due to inadequate therapy were excluded. Results:
Based on the new criteria we identified 10 patients with TS (mean
Conclusion: We have identified two subtypes of APS patients with and age 25 yrs; 8 female). Seven presented with 1 venous TE, 3 with
without valvulopathy with defining differential clinical features and cerebral sinus thrombosis, 2 with stroke/TIA, 2 with peripheral arterial
with little crossover in valvular involvement over a long follow-up TE, 2 with intra-abdominal TE, and 1 with HELLP. Only 2 patients
period, giving a high prognostic value to the initial echo assessment. had a history of APS, and 8 had no prior TE. Pregnancy was the most
common ‘trigger’ event. All patients received anticoagulant therapy
Disclosure: Nothing to disclose.
and 3 received immunosuppression. Nine survived the acute event
but 2 required amputations. Two patients developed recurrent TE
B089 1 year later. Conclusions: These patients demonstrate the clinical
spectrum of TS. We are prospectively targeting patients with TS to
A 30-year old male with ST-elevation myocardial infarction and characterize subsequent clinical course and search for novel genetic
spontaneous echo contrast in the left ventricle: A case report mutations that account for the severe phenotype of TS.
Wang H1, Khalife W1 Disclosure: Nothing to disclose.
1
UTMB, Galveston, TX

Introduction: We report a case of possible antiphospholipid syndrome B091

presented as premature Myocardial Infarction (MI) with left ventricu-
lar spontaneous echo contrast. MI and other thromboembolic events
are among the manifestations of Antiphospholipid Syndrome (APS).
Anti- b2 glycoprotein 1 antibody levels are elevated in patients with
MI, especially in the younger group, and identified as an independent
risk factor. Spontaneous echo contrast (SEC) in the atria is associated
with increased risk of systemic and pulmonary thromboembolism. Left
atrial SEC was documented in patients with APS, while left ventricular
SEC has not been reported in patients with MI and possible APS. Case
presentation: A 30-year old African male without significant risk fac-
tors for coronary artery disease except for a 5 pack-year smoking
history, presented with chest pain with ST-elevation in the inferolateral
leads on EKG. Coronary angiography showed normal right and left
circumflex coronary arteries. Interestingly, there was a thrombus in the
left anterior descending (LAD) coronary artery with distal emboliza-
tion. Echocardiography showed SEC in the left ventricle (LV) with LV
ejection fraction of 35-40%. Anticardiolipin antibody levels were
normal but anti-b2 glycoprotein 1 antibody (IgG) level was elevated.
Patient underwent aspiration thrombectomy and stent placement in
the LAD. He was discharged home on optimal medical treatment
and the anti-b2 glycoprotein 1 antibody levels were to be rechecked
in 12 weeks to confirm the diagnosis of APS. Conclusion: The patient
with premature MI and left ventricular SEC may have underlying
The prevalence of SLE, Anti-nuclear Antibodies and Antiphospholipid
Antibodies in patients with Thrombotic Thrombocytopenic Purpura
(TTP)
Vickie McDonald V1, Scully M1, Machin S1
1
UCL London, United Kingdom
TTP may be 1 or 2 and is often an autoimmune disease associated
with anti-ADAMTS13 antibodies. It is reported to be associated with
SLE and the ADAMTS13 axis may be affected by the presence of
antiphospholipid antibodies (APL). We ascertained the prevalence of
SLE, anti-nuclear antibody (ANA) positivity and antiphospholipid
antibody positivity in TTP patients. TTP episodes recorded in the
SE-London TTP registry and the UK national TTP registry were ana-
lysed from 2003-2009. There were 285 episodes of which 156 (55%)
had ANA and 124 (44%) had APL tested at diagnosis. 19% were ANA
positive, 6.5% were APL positive and 4.5% had either pre-existing
SLE or were subsequently diagnosed with SLE. Presenting clinical
features in those with SLE were: neurological 100%, fever 50%,
abdominal 16%, cardiac 33% and renal 33%. The median Hb, platelet
count, LDH, ADAMTS13 activity and anti ADAMTS13 antibody
were: 62g/L, 10 109/L, 2456IU/L, <5% and 38%. The median
number of plasma exchange (PEX) to remission was 29 and there

Lupus

were no deaths. The presenting features in those who were ANA posi-
tive/negative were not significantly different. There was no difference
between median presenting Hb (73g/L vs 88g/L), plt (13 vs 16 109/L),
LDH (1583 vs 1804IU/L) and anti-ADAMTS13 IgG (46 vs 39%,
normal <4%) in those with/ without ANA at presentation. The
median ADAMTS13 activity in both groups was <5% at diagnosis
(normal >66%) and the number of PEX to remission (17 in ANAþ/ 15
in ANA) was similar. Only 8 pts tested positive for APL. There was
no difference in the presenting clinical features or laboratory results
between those with/ without APL antibodies at presentation. The pre-
sence of APL did not affect the number of PEX to remission (16.5 in
APLþand 17 in the APL). In conclusion, in a large cohort of patients
with acquired TTP, those with underlying SLE represent only a small
percentage. The presence/ absence of ANA or APL does not appear to
affect presenting features, outcome or response to treatment. Patients
with SLE may require more PEX than those without however these
were small patient numbers and the influence of ethnicity should be
taken in to account.
Disclosure: Dr V McDonald is sponsored by an unrestricted educa-
tional grant from Baxter UK. This presentation was supported in part
by a research grant from the Medical Research Council (MRC) UK.
B092
The Role of Antiphospholipid Antibodies in Occurrence of Cardiac
Manifestations in Patients With Antiphospholipid Syndrome
Stojanovich L1, Djokovic A1, Marisavljevic D1
1 n¼5, venous n¼10, recurrent thrombosis n¼1) during the study
Clinical Center Bezanijska kosa, Belgrade, Serbia and Montenegro
Antiphospholipid syndrome (APS) patients suffer from various cardiac
manifestations. APS may manifest itself as a primary disease (PAPS)
or as a secondary disease, most commonly in the context of Systemic
Lupus Erythemathosus (SLE) This is the demonstration of the initial
results from the Serbian National APS Registry. We analyzed 256
patients:162 PAPS patients(94 female and 68 male, 45.212.9 years)
and 94 SLE patients with secondary APS (83 female and 11
male,43.115.4 years); antiphospholipid antibodies(aPL)analysis
included analysis of aCL(IgG,IgM),b2GPI and LA. In all patients
echocardiography study was performed and data considering acute
myocardial infarction, unstable angina, coronary artery bypass graft-
ing (CABG) or percutaneus coronary artery angioplasty (PTCA) and
chronic or acute heart failure were also collected. In our study group
there was 39.5% pts with aCLIgG antibodies, 58.2% with
aCLIgM36.3% with b2GPI IgG,49.9% b2GPI IgM and 52.3%with
LA. There was statistically significant difference considering presence
of aCL IgM antibodies and CABG/PTCA. Total of 77.8% APS pts
with this intervention were aCL IgM negative comparing to 1.3%of
aCLIgM positive patients with CABG/PTCA (p¼0.026). Presence of
aCL IgM was significantly higher in pts with pseudoinfective endocar-
ditis (p¼0.037). Only 3.0% LA positive patients had this cardiac man-
ifestation, comparing to 10.7%LA negative (p¼0.014). Statistically
significant positive correlation was observed considering cardiac man-
ifestations and aCLIgG titer (p¼0.01). We found that level of aCL IgG
(p¼0.027) and b2GPI IgG (p¼0.000) were in positive correlation with
valve thickening and dysfunction. Higher titer of aCL IgG in our study
group also brought higher incidence of vegetations (p¼0.007). On the
other hand, we observed negative correlation between manifestations
of coronary artery disease and aPL but without statistical significancy.
Our study showed that APS aCL IgM negative patients are less prone
to development of coronary artery disease. Higher titers of aCL IgG
pronounce the risk of valvular involvement.
Disclosure: Nothing do disclose.
Abstracts
525
B093
Epidemiology, Clinical Features and Outcome in 131 Children with
Positive Antiphospholipid Antibodies
Rozic M1, Trampus Bakija A1, Rener Primec Z1, Kitanovski L1,
Kveder T2, Avcin T1
1
Children’s Hospital, UMC Ljubljana, Slovenia; 2University Medical
Center Ljubljana, Ljubljana, Slovenia
Objective: To evaluate the prevalence of thrombotic and non-throm-
botic clinical manifestations and long-term outcome in a large unse-
lected group of children with positive antiphospholipid antibodies
(aPL). Methods: A single-center, retrospective study with longitudinal
follow-up of all children who tested positive for at least one aPL sub-
type (aCL, anti-b2GPI and/or LA) between January 1997 and January
2008 at the University Children’s Hospital Ljubljana. aPL testing was
considered in patients with autoimmune diseases, coagulation abnorm-
alities, hematological, skin and neurological disorders as well as in
infants born to mothers with aPL-positive autoimmune diseases.
Results: A total of 131 patients were enrolled, 81 girls and 50 boys
with mean age at disease presentation 9.9  5.8 years. Eighteen (14%)
were infants born to aPL-positive mothers and 113 (86%) were older
than 1 year. Mean follow-up period was 4.8  3.6 years. During the
follow-up hematological manifestations occurred in 25% (thrombocy-
topenia n¼19, leucopenia n¼5, AIHA n¼5, Evans syndrome n¼4),
non-thrombotic neurological manifestations in 24% (migraine n¼18,
seizures n¼6, chorea n¼2) and skin disease in 14% of our patients
(Raynaud’s phenomenon n¼10, livedo reticularis n¼8). In total,
11% of patients with positive aPL had thrombotic events (arterial
period. Conclusion: In our unselected cohort, thrombotic events
occurred only in 11% of all children with positive aPL evaluated at
the tertiary care pediatric hospital. The most common non-thrombotic
manifestations found in children with aPL were thrombocytopenia,
migraine headache, Raynaud’s phenomenon and livedo reticularis.
Disclosure: This presentation was supported in part by a grant from
the Slovenian Research Agency (#L3-0624).
B094
Anti-collagen antibodies and heart valve disease in patients with
antiphospholipid syndrome
Corte´s-Hernández J1, Castro J1, Torres-Salido MT1, Pardos J1,
Balada E1, Ordi-Ros J1
1
Vall d’Hebron Hospital Institute Research, Barcelona, Spain
Background: Heart valve disease is common in patients with APS. The
mechanism of valve injury is not well known. Valve histological studies
have demonstrated the presence of large amounts of collagen type I
(CI) and type IV (CIV). This collagen is likely to be an autoimmune
target as it occurs in patients with rheumatic fever. Objective: To
determine the presence of antibodies to CI and CIV as markers of
heart valve disease in patients with APS. Patients and Methods: 76
patients with primary APS, nineteen of which (25%) had valvulopathy
and 33 controls were studied. Anti-collagens I and anti-collagen IV
antibodies were measured by ELISA. Results: Antibodies to CI and
CIV were higher in patients with APS when compared to controls
(p¼0.065 vs. p¼0.092, respectively). Patients with valvulopathy had
significantly more antibodies to CI than those patients without (37%
vs. 14%, p¼0.0523). For anti-CIV antibodies, we did not find any
significant differences. This proportion increased when we analyzed
the total response to any anti-collagen antibody, either CI or CIV, in
patients with valvulopathy (58% vs. 22%, p¼0.0032). Conclusion:
Antibodies to collagen type I are present in patients with APS and
Lupus
 Abstracts

526
valvulopathy and could be a useful serologic marker of this condition.
Further studies are needed to establish the role of anticollagen anti-
bodies in the pathogenesis of heart valve disease.
Disclosure: Nothing to disclose.
disease, atrial fibrillation, hyperomocisteinemia and oral contraceptive
intake. In the group with NM we found that one or more RF were
present in 109 pts (87.2%), while only 36 pts (65.4%) without NM
presented RF (p¼0.007 chi square test). The different distributions of
RF in two groups of pts are shown in figure 1.

B095

Fetal inferior vena cava and renal vein thrombosis associated with
antiphospholipid antibodies in the mother: a case report and literature
review
Tomowiak C2, Devignes J2, Jellimann JM3, Droulle P3, Wahl D1,2,
Ab der Halden Y3, Andre JL2, Lecompte T1, Thiebaugeorges O3
1
UHP, Vandoeuvre, France; 2CHU, Vandoeuvre, France; 3Maternite,
Nancy, France

We report a case of fetal inferior vena cava and renal vein thrombosis
associated with antiphospholipid antibodies in the mother in a 36-year-
old woman with anti-phospholipid antibodies (APL), thrombosis was
extended to the right atrium, the left and right renal veins associated
with a bilateral adrenal haematoma. At 37 weeks, a caesarean section
was performed. Unfractionated heparin, then low molecular weight
heparin treatment was administrated to the newborn with a favorable
outcome. Sixteen cases of fetal inferior vena cava were identified
through a Medline search. Fetal inferior vena cava and renal vein
thrombosis can be diagnosed incidentally or in a context of fetal dis-
tress or hydrops fetalis. At an early stage of thrombosis, prenatal
ultrasound can reveal enlarged and hyper echogenic kidney with loss
of cortico-medullary differentiation. Lace-like calcifications may occur
later. As maternal thrombophilia is often present (85 % in idiopathic
cases), it is advised to perform such a diagnostic workup. However
only a minority of cases were associated with antiphospholipid anti-
bodies A sonographic follow up is necessary and an elective delivery is
to be discussed in case of fetal distress, or after 34 weeks to enable
neonatal treatment when the thrombosis is extensive as in pour case.
Prenatal thrombosis of inferior vena cava and/or renal vein can be
related to maternal thrombophilia such as antiphospholipid antibo-
dies. As fetal distress or hydrops fetalis may occur, a close monitoring
is mandatory. General therapeutic recommendations cannot be made
from the literature due to the paucity of cases. Thus the report of
individual cases may bring new insights to the management of such
thromboses.
Disclosure: Nothing to disclose.
B096
Evaluation of neurological manifestations (nm) in 180 patients with
primary APS (PAPS)
Casu C1, Andreoli L1, Fredi M1, Tincani A1
1
University of Brescia, Brescia, Italy
Objectives: To retrospectively describe the neurological manifestations
(NM) recorded in a cohort of 180 patients (pts) with PAPS. To verify
whether NM in PAPS pts are associated to particular risk factors (RF).
Methods: The study included 180 pts (23 male, 157 female) classified as
PAPS according to the revised classification criteria. The mean age at
the time of diagnosis was 37.6 y, the mean follow-up period was 4.9
yrs. Results: NM had occurred at least once in 125 pts (69.4%). NM
were recorded in 105 pts (84%) at disease onset, while the other 20 pts
(16%) suffered from new NM within the first 5 yrs from the diagnosis.
Neurological events occurred in 96 pts (76.8%) of cases before the age
of 40. We grouped the pts upon the presence or absence of NM. In
these two groups we evaluated the correlation with cardiovascular RF
such as hypertension, diabetes, dislypidemia, smoking, valvular heart
Conclusion: NM in PAPS pts occurred more frequently at disease onset,
and below the age of 40, underlying that they are part of the clinical
picture and not a consequence of chronic illness. Among the cardio-
vascular RF, hypertension and valvular heart disease were found more
prevalent in pts with NM (p¼0.0038 hypertension, p¼0.007 vascular
heart disease). Since these RF are deeply involved in the pathogenesis
of cerebrovascular disease, our findings confirm that the assessment of
RF is mandatory in PAPS pts.
Disclosure: Nothing to disclose.
B097
The value of white matter hyperintensity lesions in patients with
antiphospholipid syndrome and systemic lupus erythematosus
Sanna G1, Gordon P2, Almeida A3, Hughes GRV4, Hunt B3,4,
Khamashta MA1,4
1
Lupus Unit, Guy’s and St Thomas’ NHS Foundation Trust, London,
United Kingdom; 2Department of Rheumatology, King’s College
Hospital NHS Trust, London, United Kingdom; 3Department of
Hematology, Guy’s and St Thomas’ NHS Foundation Trust, St Thomas’
Hospital, London, United Kingdom; 4Lupus Research Unit, The Rayne
Institute, King’s College London School of Medicine, London, United
Kingdom
Background: Silent brain infarcts and white matter lesions are fre-
quently seen on MRI in APS and SLE and their significance is
matter of debate, as it is their association with aPL. We studied the
prevalence of white matter hyperintensity lesions (WMHL) and stroke
in a population of patients with APS and SLE. Methods: Out of 193
patients with CNS involvement, 52 patients had primary APS (PAPS)
and 141 SLE (48 with secondary APS (SAPS) and 93 aPL negative
(SLE only)). Results: WMHL were found in 22/52 patients with PAPS,
28/48 with SAPS and 26/93 with SLE only. WMHL were more fre-
quent in patients with APS and SAPS when compared to those without
APS or SLE only (p¼0.002 and p¼0.0009, respectively). There were no
differences in the frequency of WMHL between PAPS and SLE only
(p¼0.09) and between PAPS and SAPS (p¼0.16). Evidence of large
vessel disease (i.e. classical ischemic stroke) was found in 32/52 patients

Lupus

with PAPS, in 22/48 patients with SAPS, and in 6/93 patients with SLE
only. These lesions were more frequent in PAPS and in SAPS than in
SLE only (p<0.0001 and p<0.0001). Conclusion: WMHL are a promi-
nent feature of APS. Targeted therapy may be beneficial for these
patients.
Disclosure: Nothing to disclose.
B098
Migraine and anti-phospholipid antibodies: a case-control study
Bazzan M1, Cavestro C2, Micca G2, Di Pietrantonj C3, Roccatello D1
1
CMID - Giovanni Bosco Hospital, Turin, Italy; 2Headache Unit -
S.Lazzaro Hospital, Alba, Cuneo, Italy; 3Regional Service for
Epidemiology, Alessandria, Italy
Background: Migraine physiology is still poorly understood; blood
hypercoagulability was hypothesized to be involved. Presence of anti-
phospholipid antibodies is an acquired, immune-mediated, strong risk
factor for developing both venous and arterial thrombosis. At our
knowledge, a wide, standardized, case control study, designed to eval-
uate prevalence of antiphospholipid antibodies in migraine patients, is
lacking. Patients and Methods: 284 patients (59 men, 225 women) with
migraine diagnosed according to the last International Headache clas-
sification, were consecutively enrolled from September 2008 up to
August 2009 in our outpatients Headache Clinic. In the same period
227 age and sex matched subjects not suffering from any type of head-
ache, were enrolled as control group. All patients and controls were
evaluated for lupus anticoagulant, anti-cardiolipine antibodies, and
anti-b2GPI. All positive (according to ‘‘Sydney’’ criteria) APLs
patients and controls were re-tested after at least 12 weeks. Analysis
of variance per groups was performed, using SAS software. Results: 41
out of 284 (14,4%) migraine patients (7 with LAC positivity, 25 anti-
beta2GP1 and 9 ACA) and 8 out of 227 (3,5%) controls (no LAC, 6
anti-beta2GP1 positivity, 2 ACA) were APLs positive, confirmed after
12 weeks. The statistical differences between the two groups were:
LAC p¼0.006; OR 8.52 (CI 1.07-67.41), anti-b2GPI p¼0.004; OR
3.51(1.41-8.71); ACA¼ n.s. Conclusions: A significantly higher preva-
lence of APLs was found, and confirmed, in migraine patients com-
pared to the non-migraine control group. Our data show that APLs
could have a pathogenetic role in migraine, at least in a subset of
patients.
Disclosure: Nothing to disclose.
B099
Renal failure, steno-obstructive abnormalities and femoral superficial
artery dissection in a young Caucasian male with antiphospholipid
syndrome
Cianci R1, Gigante A1, Barbano B1, Lai S1, Gasperini ML1, Ascoli G1,
Amoroso A1
1 Lupus patients had lower mean (SD) performance on all ANAM
University of Rome, Rome, Italy
A 22-year-old Caucasian male was hospitalized for intermittent clau-
dication, hyposphygmia of right lower limb, hypertension and dete-
rioration of renal function. CDU showed the presence of a high grade
stenosis of right renal artery and an atrophic left kidney; lower extre-
mity CDU revealed occlusion of right external iliac artery and dissec-
tion of left superficial femoral artery. Laboratory studies showed
positivity for LAC, prolonged PTT, hyperhomocysteinemia without
MTHFR mutation, positivity for anticardiolipin IgG antibodies, anti-
nuclear antibodies and negativity for ANCA. Few days later, the
patient presented with an acute kidney injury (serum creatinine: 3.36
mg/dl; serum Kþ: 5.9 mEq/L). We performed an angiography study
Abstracts
527
and renal artery stenosis was treated with renal percutaneous trans-
luminal angioplasty and stenting. Angio-CT showed patency of right
renal artery stenting, atrophic left kidney and many steno-obstructive
disorders in splanchnic vascularization such as ectasia of celiac tripod,
obstructed origin of right hepatic artery, reduced caliper of splenic
artery and obstructed origin of superior mesenteric artery with a hyper-
trophic Riolano arcade as collateral circles.A renal biopsy showed
istopathological findings of APS-nephropathy. About 2 months later
after PTA, a CDU scan revealed the presence of intrastent restenosis
(>80% lumen reduction) treated with Cutting Balloon Angioplasty.
The patient was treated with anticoagulant therapy for PAPS because
SLE-ACR criteria were not yet fulfilled.
Disclosure: Nothing to disclose.
B100
Relationship between cognitive performance and Antiphospholipid
Antibody levels in lupus patients and controls
Brey R1, Holliday S2, Saklad A1, Zarzabal LA1, Petri M3, Reveille J4
1
UTHSCSA, San Antonio, TX; 2South Texas Veterans Health Care
System, San Antonio, TX; 3Johns Hopkins University, Baltimore, MD;
4
UT Houston HSC, Houston, TX
The purpose of this abstract is to describe the relationship between
aCL, anti-b2GPI and antiribosomal P (anti-P) levels and cognitive
performance using a computerized neuropsychological testing instru-
ment (ANAM) in Lupus patients and matched normal control sub-
jects. ANAM was given to Lupus patients and controls at each study
visit, occurring every 6 months. Blood was also obtained at each visit
for antiphospholipid antibodies (aPL) and anti-P. A total of 187 Lupus
patients and 181 controls have been enrolled thus far. There were no
differences in age and education level between Lupus patients and
controls. There were significant associations between Lupus/Control
patients with respect to gender [Lupus: female (89%) vs male (11%),
Control: (77% vs 23%), p¼0.002] and race [Lupus: white(60%), black
(37%), and other (3%), Control: (70%, 23%, and 8%, p¼0.004).
subtests. Significant differences were seen in the following: Code sub-
stitution delayed (Lupus: 27.8  1.5, Control: 32.7  1.5, p<0.001);
Code substitution immediate (28  1.6 vs 32.6  1.5, p<0.001); Code
substitution (36.2  1.4 vs 39.3  1.4, p¼0.004); Performance (71.3 
2.6 vs 77.2  2.5, p¼0.003); Digit set (26.9  2.0 vs 30.7 1.9, p¼0.01);
Matching (22.8 1.2 vs p¼0.02); Spatial processing (18.1  1.1 vs 19.9
1.0, p¼0.02). There was no significant difference for simple reaction
time and the Sternberg. For Control subjects, there was a modest
inverse correlation between anti-b2GPI IgA levels and Code substitu-
tion delayed, Code substitution immediate, Code Substitution,
Performance, Matching, Simple reaction time, Spatial processing and
Sternberg; and between anti-b2GPI IgG and anti-P and Matching.
Lupus
 Abstracts
528
For Lupus patients, modest inverse correlations were seen between
anti-b2GPI IgA and Performance and Digit set comparison; and
between Anti-P and aCL IgA and Sternberg. This suggests that aPL
may be important predictors of poor cognitive performance in Lupus
patients and controls, but the relationship is stronger in controls.
Disclosure: This presentation was made possible by a grant from NIH/
NINDS.
B101
Detection of anti-argonaut 2 (Agi2/SU) and –Ro antibodies by
immuunoprecipitation in primary antiphospholipid syndrome (PAPS)
patients
Ceribelli A1, Cavazzana I1, Tincani A1, Franceschini F1, Chan E2,
Satoh M3
1 surveys in patients with SLE and APLA, even without a history of
Università degli Studi, Spedali Civili, Brescia, Italy; 2Department of
Oral Biology, University of Florida, Gainesville, Florida; 3Department of
Medicine, University of Florida, Gainesville, Florida
Purpose: Primary antiphospholipid syndrome (PAPS) is defined as
patients with anti-phospholipid antibodies and thrombotic/obstetric
symptoms, without systemic lupus erythematous (SLE). PAPS patients
can have SLE-like features or evolve into SLE during follow-up. The
aim of our study is to characterize lupus autoantibodies in PAPS to
analyze their clinical and laboratory correlations and the predictive
role for SLE evolution. Methods: Sera from 52 PAPS patients were
screened by indirect immunofluorescence (IIF) antinuclear antibodies
(ANA), immunoprecipitation (IP) of 35S-labeled K562 cell extract, and
ELISA [anti-Argonaute2 (Ago2, Su), 60 and 52kRo, La, dsDNA)].
Anti-Ago2/Su positive sera were also tested for anti-GW body
(GWBs) by IIF double staining using rabbit anti-GWBs serum.
Results: ANA were positive in 56% of PAPS patients. Anti-Ago2/Su
antibodies were found in 13% (7/52) and anti-Ro/SSA in 10% (5/52)
by IP. Interestingly, 79% (41/52) were negative by IP. No other lupus-
related autoantibody was detected. No particular association with clin-
ical features of anti-phospholipid syndrome was found, except that
IgG anti-beta2glycoproteinI (b2GPI) antibodies were less frequent in
anti-Ago2/Su positive patients (p ¼ 0.02). None of anti-Ago2/Su or –
Ro positive patients developed SLE during follow-up. Ago2 is a key
component of GWBs and IIF staining of Ago2 localizes to GWBs, but
only 1/7 anti-Ago2/Su serum showed a typical cytoplasmic GWBs
staining. Conclusions: Anti-Ago2/Su (13%) and –Ro (10%) antibodies
were found in sera from PAPS. No other lupus-related autoantibodies
were identified. Clarifying why Ago2/Su and Ro are specific targets in
PAPS may help to understand the mechanisms of autoantibody
production.
Disclosure: Nothing to disclose.
B102
The risk of Libman-Sacks endocarditis is strongly associated with
antiphospholipid antibodies in patients with systemic lupus erythematosus
Zuily S1, Dotto P2, Perret-Guillaume C3, Bode´-Dotto E4, de Maistre E4,
Selton-Suty C5, Bruntz JF5, Regnault V6, Eschwe`ge V4, Lecompte T4,
Wahl D1
1
Vascular Medicine unit, CHU Nancy, Vandoeuvre, France;
2
Cardiology, Saint Nicolas de Port, France; 3Geriatric Unit, CHU
Nancy, Vandoeuvre, France; 4Haematology, CHU Nancy, Vandoeuvre,
France; 5Cardiology, CHU Nancy, Vandoeuvre, France; 6Inserm U961,
Nancy University, Vandoeuvre, France
Libman-Sacks endocarditis (LBE) is a frequent feature in patients with
systemic lupus erythematosus (SLE) but the role of antiphospholipid
Lupus
antibodies (APLA) is controversial. Our objective was to assess if heart
valve disease (HVD) and especially LBE are associated with APLA in
patients with SLE. We performed a meta-analysis of primary studies
between January 1983 and october 2009 retrieved from the PubmedÕ ,
EmbaseÕ , and Cochrane LibraryÕ database. From 23 primary studies
representing 1656 patients, frequencies of HVD and APLA in SLE
patients were 30.7% and 40.3% respectively. The overall odds ratio
for LBE and HVD were 3.51 [CI 95%: 1.93-6.38] and 3.13 [CI95%:
2.31-4.24] respectively. The risk of HVD varied depending on APLA
types: lupus anticoagulant: 5.88 [CI 95%: 2.92-11.84]; aCL: 3.28
[CI95%: 2.06-5.22]; 5.63 [CI 95%: 3.53-8.97] and 1.67 [CI 95%: 0.46-
6.05] for IgG and IgM aCL respectively. Overall, the presence of
APLA in SLE patients is significantly associated with an increased
risk for LBE and HVD. The risk conferred by IgG aCL is as strong
as LA and there is no association between IgM aCL and HVD.
Clinically, this should lead to perform systematic echocardiographic
valve dysfunction. Finally, HVD and/or LBE could be considered as
potential APS criteria.
Disclosure: Nothing to disclose.
B103
Risk factors of clinical events recurrence in a cohort of antiphospholipid
syndrome patients
Belizna C1, Primard E2, Louvel JP2, Levesque H2
1
Chu Angers, Angers, France; 2Chu Rouen, Rouen, France
Introduction: Despite regular treatments, recurrence of thrombosis or
obstetrical events has been frequently reported in antiphospholipid
syndrome patients (APS). Previous data came from studies performed
before the new criteria of antiphospholipid syndrome. Purpose:
Therefore, we performed a five year longitudinal cohort study in
order to search for risk factors for clinical events recurrence, according
to new criteria for APS. Methods: 59 APS patients mean age 36yo,
were prospectively studied for 5 years. All the new clinical manifesta-
tions recorded after the diagnosis of APS, such as thrombosis or obste-
trical complications and which appeared under regular treatment were
considered such as recurrent event. Results: 27 new events were
recorded in 19 patients (32%). Two patients failed to fulfill the
study. The patients with anti b2GPI antibodies disclosed a high inci-
dence of thrombosis and/or obstetrical events (47%), as well as
patients with IgG anticardiolipin antibodies (45%). A multivariate
regression analysis revealed that anti b2GPI antibodies were an inde-
pendent risk factor for clinical events recurrence in APS patients (odds
ration 9.75; 95% confidence interval, 1.94 to 49.36). Conclusion: anti
b2GPI antibodies represent the strongest predictor of new clinical
events in our APS patients.
Disclosure: Nothing to disclose.
B104
Prevalence and clinical usefulness of the antiphospholipid and
anti-cofactor antibodies in different preeclampsia subsets in a Spanish
cohort
Ferrer-Oliveras R1, Garcı´a Ruiz I1, Zarzoso C1, Rodrigo-Anoro MJ1,
Cabero-Roura L1, Alijotas-Reig J1
1
Hospital Universitario Vall D’Hebron, Barcelona, Spain
Aim: To study the prevalence and clinical correlations between
different aPL and PE. Patients and Methods: Prospective, cohort
study. Setting: Vall D’Hebron University Hospital. Subjects: Study
group: 99 women with PE; control group: 82 healthy pregnant.

Methods: We studied different aPL using commercial ELISA kits:
aCL, antib2GPI, aPS, antiANXA5 and lupus anticoagulant (LA).
LA was analyzed using the standard recommended coagulometric
assay. Women referred as a aPLþ, had two positive test 12 weeks
apart minimum. Blood samples were taken during the clinical compli-
cation or within the 12 following weeks. Results: aPL prevalence was
14.14% in study group vs. 7.23% in control group. Excluding
antiANXA5 positive women, aPL prevalence was 13.19% vs. 3.61%
(p¼0.034). Analyzing differences in severe PE subsets vs. controls, we
obtained the following results: for two or more aPL positivity: 9.09%
vs. 1.20% (p¼0.037); aCL-IgM 10.91% vs. 1.20% (p¼0.016);
antib2GPI IgG/IgM 10.91% vs. 1.90% (p¼0.016); antib2GPI IgM
9.09% vs. 1.20% (p¼0.037); aPS 7.27% vs. 1.20% (0.082).
Comparing early-onset PE vs. controls we showed: aCL-IgM 11.11%
vs. 1.20% (p¼0.029); antib2GPI IgM 8.33% vs. 1.20% (p¼0.082), and
aPS 8.33% vs. 1.20% (p¼0.082). No differences were obtained for any
aPL in mild PE and late-onset PE subsets. Conclusions: aPL prevalence
in PE women is twice than in healthy pregnant. IgM isotypes are better
markers than IgG. IgM-aCL and IgM-antib2GPI are good biological
markers for early-onset and severe PE. The presence of multi-positive
test for aPL positivity seems to be a useful risk marker for severe and
early-onset PE.
Disclosure: Nothing to disclose.
B105
CAPS and SIRS
Makatsaria A1, Khhizroeva J1
1
Moscow Sechenov Medical Academy, Moscow, Russian Federation
Objective: Catastrophic APS presents very severe form of APS result-
ing in multiple microvascular thrombotic events, of rapid onset, and
Abstracts
529
causing multiorgan failure, a picture suggestive of septic shock, in
which, there is a massive, acute inflammatory response. Materials
and methods: we examined 21 patients with CAPS from our practice.
All patients were screened for genetic thrombophilia and antiphospho-
lipid antibodies. Results: the triggering events for the development of
CAPS were severe preeclampsia in 3 patients, HELLP-syndrome in 2
patients and infection (pneumonia, septicemia) in 4 patients. In 2 of
these women multiorgan failure precipitated by severe preeclampsia
complicated with placental abruption was fatal despite intensive treat-
ment (anticoagulants - LMWH, corticosteroids, plasma exchange). In
other 15 cases conducted therapy was successful. Multigenic thrombo-
philia was identified in 88% cases. Thrombophilia included prothrom-
bin gene G20210A mutation (11,7%), MTHFR C677T (29,4%),
4G/5G PAI-1 polymorphism (47%), -455G/A fibrinogen (41,2%),
t-PA I/D (23,5%), deficiency of protein C (11%). In all patients we
found acute or subacute wavelike disseminated intravascular coagula-
tion syndrome (DIC). Conclusion: CAPS bring together systemic syn-
dromes like APS, DIC and systemic inflammatory response syndrome.
Thrombophilia might be an important trigger factor for CAPS by
predisposing to procoagulant and proinflammatory state. Coexisting
of APS and multigenic thrombophilia represents the most unfavorable
condition, which determines a high risk of obstetric complications and
CAPS. CAPS might be more frequent than it was previously thought.
The development of obstetric complications in patients with APS
makes it necessary to suppose presence of the catastrophic variant of
this syndrome. LMWH is essential for prophylaxis and treatment of
CAPS due to its anticoagulant and anti-inflammatory activities.
Disclosure: No response indicated.
Lupus
 Abstracts

530

Abstracts presentations Friday April 16th, 2010

C106 C107

Real world experience with Antiphosphatidylserine Antibodies (aPS) Phosphatidylserine-Dependent-Antiprothrombin Antibody IgM
Azar L1, Erkan D2 (IgM-aPS/PT) Induces to Early Pregnancy Loss by Microcirculatory
1
Beth Israel Medical Center, New York, NY; 2Hospital for Special Disturbance
Surgery, New York, NY Yamazaki M
Kanazawa University Graduate School of Medicine, Kanazawa,
Background: aPS have not been included in the Updated Sapporo Ishikawa, Japan
Classification Criteria as the aPS assay requires better standardization
and the clinical/prognostic significance of aPS requires further Objectives and Methods: Pregnant morbidity, one of clinical manifesta-
research. However, in a real world setting, aPS are occasionally tions of antiphospholipid syndrome (APS), is classified into recurrent
ordered as part of the antiphospholipid syndrome work-up. spontaneous abortions (RSA) during early pregnancy and intrauterine
Objective: To descriptively analyze the laboratory and clinical charac- growth retardation (IUGR) or intrauterine fetal distress (IUFD) in
teristics of aPS-tested patients included in our aPL/APS registries. mid to late pregnancy. Of these, it is considered that IUGR and
Methods: We systemically reviewed the characteristics of aPL-positive IUFD may be induced by placental thrombi, although, the pathogen-
patients (lupus anticoagulant test, anticardiolipin antibodies IgG/M/A eses of early spontaneous abortions have not been clarified yet in APS.
> 20U, and/or anti-Beta-2-glycoprotein-I antibodies IgG/M/A > 20U Our recent study revealed that IgM phosphatidylserine dependent anti-
tested twice at least 12 weeks apart) who were also tested for aPS. prothrombin (IgM-aPS/PT) is frequently shown in polyarteritis
We grouped patients as: a) aPS positive (>20U); and b): aPS negative. nodosa patients with microthrombi-induced livedo reticularis is and
We compared the two groups using the Fischer’s exact test. Results: skin ulcers (Arthritis Rheumatism. 57: 1507, 2007 & 59, 561, 2008,
The characteristics of aPS-positive (n: 11) and aPS-negative (n: 6) Arch Dermatol, 145: 171, 2009). From these observations, we hypothe-
patients are shown below. All patients were tested for aPS once sized that spontaneous abortions in APS patients may be induced
except one patient; the highest aPS isotype was IgG, IgM, and IgA thrombi in chorionic blood vessels, and measured several kinds of
in 5 (46%), 5 (46%%), and 1 (9%), respectively. aPS antibodies were antiphospholipid antibodies (aPLs) in consecutive 112 patients with
significantly more common in patients with vascular events (p¼0.002), recurrent spontaneous abortions without any well-known etiologies.
arterial thromboses (p¼0.03), or neurologic events (cerebral vascular Results: Eighty-nine (79.4%) of 112 patients with RSA have IgM-
accidents and transient ischemic accidents (p¼0.04)). No significant aPS/PT with high titers compared with 1 of 100 normal pregnant
difference was noted when we compared the aPL profiles and non- women (OR¼15.4, p < 0.001). In contrast, the other antiphospholipid
criteria aPL manifestations of the two groups. antibodies including anticardiolipin antibody-IgG and –IgM, and IgG
aPS/PT were not significant increased in patients with RSA.
Immunopathological stains revealed that IgM-aPS/PT bound to arter-
aPS(þ), N:11 aPS(-), N:6 p iolar and venular endothelial cells. Conclusion: These findings sug-
gested that IgM-aPS/PT is an independent risk factor of RSA. It is
Demographics
considered that microcirculatory disturbance of chorion membrane
Female 9 (82%) 6 (100%) 0.51
due to IgM-aPS/PT may be one of major etiologies of RSA.
Age (meanSD), years 420.1 403.0 0.76
Antiphospholipid Syndrome 7 (63%) 0 0.03 Disclosure: Nothing to disclose.
Asymptomatic aPL 2 (18%) 6 (100%) 0.002
Catastrophic APS 2 (18%) 0 0.51
Associated SLE/RA 3 (27%) 3 (50%) 0.60 C108
Criteria Events
Vascular Events 9 (81%) 0 0.002 Association between antiphospholipid and anticardiolipin antibodies in
Venous Thrombosis 3 (27%) 0 0.51 patients with pregnancy complications
Arterial Thrombosis 7 (63%) 0 0.03 Otero A1, Lena A1, Attarian D1, Mota N1, Domı´nguez G1, De Los
Stroke and/or TIA 6 (54%) 0 0.04 Santos S1, Pita D1
1
Pregnancy Events 4/9 (44%) 0 0.10 CEAHT, Montevideo, Uruguay

Antiphospholipid antibodies (APA) are a group of heterogeneous anti-

Conclusion: Despite small numbers, our results suggest that among
patients with comparable aPL profiles, those who test positive for
aPS antibodies are at higher risk of aPL-associated vascular events.
These results should stimulate our efforts towards a better standardi-
zation of the aPS assay and eventually a judicious wider use of the test
to further clarify its clinical and prognostic significance.
Disclosure: Nothing to disclose.
bodies directed against phospholipids-protein complexes, with clinical
characteristics associated to thrombosis and repetitive pregnancy fail-
ure. The assay to study APA includes cardiolipin among the phospho-
lipids against which the presence of antibodies is investigated.
Anticardiolipin antibodies (ACA) are the antibodies which have pre-
sented the most important clinical significance among the whole APA.
Objectives: The aim of this study is to determine the association
between APA and ACA presence in patients with pregnancy

Lupus

complications. Methods: 148 patients between 21 and 43 years old,
who were studied for the presence of APA and ACA, because they
presented pregnancy complications associated to antiphospholipid
antibodies. The methodology employed was ELISA for APA screening
assay and IgG and IgM ACA isotypes. The presence or not of associa-
tion between APA and ACA isotypes was statistically studied by Chi
Square. Results: 129 positive results were obtained for APA and 95 for
ACA (29 IgG isotype and 66 IgM isotype) X2¼ 1.98 between APAþ
and ACA IgGþ; X2¼ 7.47 between APAþ and ACA IgMþ
Conclusions: Since X2 0.05 is 3.84 (1 freedom grade), there is no associa-
tion between APAþ and ACA IgGþ, but there is association between
APAþ and ACA IgMþ. A greater number of patients resulted APAþ
than ACAþ. It may be due to the fact that APA includes antibodies
against a greater panel of phospholipids which includes cardiolipin
among others. ACA IgMþ isotype which is more frequently found
in these patients presents association with APAþ. However the lack
of association between ACA IgGþ and APAþ, confirms the conveni-
ence of performing both assays, APA and ACA isotypes, excluding
neither of them in patients with pregnancy complications.
Disclosure: Nothing to disclose.
C109
Multiple (triple or quadruple) aPL positivity is much more prevalent in
patients with definite antiphospholipid syndrome
Forastiero R1, Martinuzzo M2, Iglesias Varela ML2, Cerrato G2
Background: According to the 2006 criteria for the APS, the laboratory
diagnosis requires the presence of at least 1 of the 3 common aPL (LA,
IgG/IgM aCL and anti-b2GPI). The detection of persistently elevated
levels of aPL on 2 occasions at least 12 weeks apart and high aPL titers
(> 40 GPL/MPL for aCL and > the 99th percentile for anti-b2GPI) are
other requisites. It is advised to classify APS patients as category I (any
combination of aPL), IIa (LA alone), IIb (aCL alone) and IIc (anti-
b2GPI alone). The clinical criteria are type 1 (vascular thrombosis) or
type 2 (pregnancy morbidity classified as 2a/2b/2c). The inclusion of
antiprothrombin antibodies (anti-PT) in the APS criteria is still pre-
mature. Objective: To classify the patients referred to our centre that
tested positive for aPL during the last 4 years. Methods: LA was
determined according to ISTH guidelines and aCL/anti-b2GPI/anti-
PT by ELISAs. Titers were positive when aCL >40 units and anti-
b2GPI and anti-PT >20 arbitrary units. Results: There were 187
patients with confirmed positive aPL (LA, aCL and/or anti-b2GPI).
The diagnosis of APS was made in 97 patients (66 type 1, 10 type
1þ2a/b/c, and 21 type 2a/b/c). The other 90 patients had no clinical
criteria of APS (non-APS) (asymptomatic, SLE, thrombocytopenia,
neoplasia, <3 abortions). Anti-PT were found in 33/97 (34%) patients
with APS and in 15/90 (17%) with non-APS.
I IIa IIb IIc
APS 61 (62.9%) 5 (5.2%) 20 (20.6%) 11 (11.3%)
non-APS 32 (35.5%) 23 (25.6%) 30 (33.3%) 5 (5.6%)
The combination of aPL was more prevalent in definite APS than in
non-APS (p¼0.0002, OR 3.1, 95%CI 1.7-5.6). The triple positivity for
LA/aCL/anti-b2GPI was present in 35 patients with APS and in 12
with non-APS (p¼0.0004, OR 3.7) and the quadruple positivity for
LA/aCL/anti-b2GPI/anti-PT in 26 with APS and in 4 with non-APS
(p<0.0001, OR 7.9). Conclusions: The risk of APS-related clinical
features progressively increases with the number of positive aPL
Abstracts
531
tests. The multiple (triple or quadruple) aPL positivity distinguish
high-risk patients for thrombosis.
Disclosure: Nothing to disclose.
C110
The diagnostic significance of Antiphospholipid-Score (aPL-S) for
antiphospholipid syndrome (APS) and the predictive value of aPL-S
for thrombosis
Otomo K1, Atsumi T1, Fujieda Y1, Kato M1, Amengual O1, Oku K1,
Kataoka H1, Horita T1, Yasuda S1, Koike T1
1
Hokkaido graduate school of medicine, Sapporo, Hokkaido, Japan
Purpose: To evaluate the efficacy of aPL-S for the diagnosis of APS
and predictive value for the development of thromboses in autoim-
mune diseases Method: 1) Five clotting assays (APTT and dRVVT
(mixing and confirm tests) and kaolin clotting time) and 6 ELISAs
(IgG/M anticardiolipin, IgG/M anti-beta2GPI and IgG/M phosphati-
dylserine dependent antiprothrombin antibodies) were performed in 233
patients with autoimmune diseases. The aPL-S was compared with the
history of thrombosis/pregnancy morbidity. 2) The second study com-
prised 441 patients with autoimmune diseases and 302 (73.5%) were
followed-up with a mean duration of 67 15 months. All patients per-
formed APL tests and the aPL-S was compared with the newly devel-
oped thromboses during the follow-up period. Results: 1) The aPL-S
was higher in patients with thrombosis/pregnancy morbidity (n¼46)
than in those without (p¼0.00001). 2) Thirty-two patients newly devel-
oped thromboses. Twelve of 39 patients with aPL-S >30 developed
thromboses. Patients with aPL-S >30 had a higher risk of thromboses
than those without (Odds-Ratio: 5.40[95%CI: 2.38-12.2]). Conclusion:
The aPL-S is quantitatively useful for diagnosing APS, and may be a
predictive marker of thrombosis.
Disclosure: Nothing to disclose.
C111
Analytical and clinical performance of a new, fully automated,
chemiluminescent assay panel for the diagnosis of antiphospholipid
syndrome
Morin K2, de Moerloose P3, Musial J4, Arnout J5, Ruiz N1
1
Biokit, Lliçà d’Amunt, Barcelona, Spain; 2Instrumentation Laboratory,
Bedford, MA; 3University Hospital, Geneva, Switzerland; 4Jagiellonian
University, Krakow, Poland; 5University of Leuven, Leuven, Brabant,
Belgium
The analytical and clinical performance of a new assay panel for aPL
antibodies, [HemosILÕ AcuStar aPL, Instrumentation Laboratory,
(IL)] was evaluated and compared to three aPL ELISA kits (Phadia
VarelisaTM, Corgenix REAADSÕ , and INOVA QUANTA LiteTM).
The new assays detect IgG and IgM aCL and ab2GPI in serum or
citrated plasma on a fully automated, random-access, chemilumines-
cent analyzer [ACL AcuStarTM (IL)]. The cut-off of the four assays is
20U/mL, based on the 99th percentile of a normal range study; linear-
ity ranges extend in rerun 40-6000 times the cut-off. Analytical and
clinical performance were evaluated by analyzing 321 clinically well-
characterized samples with the aPL panel and the ELISA kits at three
different sites. Patient samples with autoimmune disorders and a con-
trol group were included. Results were interpreted as positive or nega-
tive using the manufacturers’ cut-off values. AcuStar aPL panel
showed 83%-96% agreement among all three kits. Slopes ranged
between 0.9-1.1, and r values were >0.997 indicating minimal interla-
boratory variation. When used to diagnose APS, the sensitivity and
specificity of the combination of the four aPL assays were, respectively,
Lupus
 Abstracts
532
72.8%, 87.3% (AcuStar), 75.0%, 74.2% (REAADS), 57.6%, 85.6%
(Varelisa) and 64.1%, 88.6% (QUANTA Lite). Odds ratios demon-
strated the ability of the assays to predict a thrombotic event or obste-
tric complication. With the combination of the four aPL assays, the
odds ratio was 6.4 (AcuStar), 5.7 (QUANTA Lite), 4.0 (REAADS),
and 3.4 (Varelisa). In conclusion, the HemosIL AcuStar panel showed
equal to superior clinical performance to the three ELISAs when used
to aid in the diagnosis of APS, and also in the prediction of clinical
complications. The new global aPL panel also provides excellent ana-
lytical performance, and quick diagnostic information, with the great
advantage of full automation.
Disclosure: This abstract was supported by Instrumentation
Laboratory.
C112
Lack of anti-annexin A5 antibodies in women with spontaneous
pregnancy loss
Garcı´a Ruiz I1, Zarzoso C1, Ferrer-Oliveras R1, Lopera AF1, Farran-
Codina I1, Rodrigo-Anoro MJ1, Casellas-Caro M1, Alijotas-Reig J1
1 Disclosure: Nothing to disclose.
Hospital Universitario Vall D’Hebron, Barcelona, Spain
Objective: The aim was to evaluate the role of anti-annexin A5 (anti-
ANXA5) antibodies as risk factor for recurrent miscarriage (RM) and
unexplained fetal loss (UFL). Patients/Methods: Retrospective, cohort
study. Setting: Vall D́Hebron University Hospital. Subjects: 122 women,
in two groups: study group: 54 women with RM/UFL; control group: 68
pregnants without RM/UFL. Intervention: Antiphospholipid, mainly
anti-ANXA5 antibody analysis. Comparison of all antiphospholipid
antibodies between groups. Results: Antiphospholipid antibody (aPL)
prevalence in study group was 10/54 (14,8%) and 5/68 (7,3%) in con-
trol group (p¼0,09). In RM subgroup, it was 3/25 and 9/34 in UFL
versus 5/68 in controls (p¼0,013). Lupus anticoagulant (LA) was pre-
sent in 4 cases, all belonging to study group (p¼0,011). 4/34 women
with UFL were positive for anticardiolipin antibodies IgG (IgG-aCL)
versus 1/68 in controls (p¼0,041). In RM subgroup, anti-ANXA5 anti-
bodies were positive in 2/25 versus 3/68 in controls, and in UFL sub-
group, 3/34 versus 3/68 cases (p¼1,000). Conclusion: According to our
results, anti-ANXA5 antibodies should not be considered as a risk
factor for RM/UFL.
Disclosure: Nothing to disclose.
C113
Antiphosphatidylethanolamine antibodies: clinical significance and
methodological aspects of their detection
Sanmarco M1
1
Hospital La Conception, Marseille, France
The nosological limits of the antiphospholipid syndrome (APS) are not
completely achieved. In particular, the diagnosis of APS cannot be
established for patients with clinical features highly suggestive of
APS in the absence of the laboratory criteria of this syndrome.
Interestingly, in such a clinical context, investigation of antiphospha-
tidyletanolamine antibodies (aPE) deserves a particular attention
because of the results from several studies showing these antibodies
as significantly associated with the main clinical events of APS, fetal
loss and/or thrombosis. Regarding obstetrical complications, aPE have
been reported to be significantly more frequent in women with unex-
plained (UFL) than in those with explained early fetal losses or in
healthy mothers. Two different studies showed aPE as a higher inde-
pendent risk factor for early UFL than anticardiolipin and anti-b2GPI
antibodies. Interestingly, aPE have also been described as the only
Lupus
aPLs found in most of the cases (73%). The relationship between
aPE and thrombosis has been also reported in several studies. In par-
ticular, in a multicenter study set up in the frame of the European
Forum on aPL, they were described with a prevalence of 15% in
unexplained venous thromboses and mainly found as the only aPLs.
In this study, IgG-aPE were found as an independent retrospective risk
factor for venous thrombosis with an odds ratio of 6.aPE are measured
by ELISA but no consensual methodology does exist at the moment.
Variations in ELISA components leading to a great inter-laboratory
variability of the results make difficult the comparison of data. Among
the different components, the buffer supplement has been reported by
several investigators as the critical factor in measurement of aPE. An
attempt to reach a consensus was carried out in the frame of the
European Forum on Antiphospholipid Antibodies and did not suc-
ceed. Because of the significant association of these antibodies with
the clinical features of APS, the definition of the optimal conditions
for a reliable test is needed. Thus, this meeting might be the opportu-
nity to compare the different experiences on aPE detection and to
propose some recommendations for decreasing the discrepancies
between the laboratories.
C114
Comparison of Clinical Performance of Commercial EIAs to Novel
Multiplexed Immunoassays for the Bio-Rad BioPlexTM 2200; An Aid in
the Diagnosis of Anti-Phospholipid Syndrome
Watkins M1, Khamashta M2, Bertolaccini M2, Tincani A3
1
Bio-Rad Laboratories, Benicia, CA; 2The Rayne Institute, St Thomas’
Hospital, London, United Kingdom; 3Laboratorio di Reumatologia e
Immunologia Clinica, Brescia, Lombardy, Italy
Purpose: Three prototype kits have been developed for the BioPlex
2200, a fully automated random access multiplex testing platform, to
detect anti-cardiolipin (CL) or anti-beta-2-glycoprotein I (b2GPI) IgG,
IgA or IgM antibodies, biological markers known to have a strong
association with Anti-Phospholipid Syndrome (APS). The purpose of
this study was to compare the ability of these multiplexed APS assays
and the corresponding commercial EIA kits to detect clinically char-
acterized APS samples while retaining specificity among a non-APS
population. Methods: The cohort of the study comprised 208 normal
(assumed non-disease) and 198 primary and secondary APS disease
state samples (kindly provided by Drs. M. Khamashta and A.
Tincani). All samples were analyzed by the BioPlex 2200 APS IgG,
IgA and IgM kits and the corresponding commercially available EIA
kits. Results: Tables 1 contains a summary of the clinical performance
of the CL and b2GPI assays. Overall, the clinical performances of
BioPlex versus the EIAs are comparable for the CL and b2GPI kits
at detecting APS disease.
BioPlex EIA #1 EIA #2
Analyte Sensitivity Specificity Sensitivity Specificity Sensitivity Specificity
CL IgG 57% 98% 60% 98% 84% 80%
CL IgA 48% 98% 8% 98% 28% 94%
CL IgM 23% 95% 18% 99% 48% 95%
b2 IgG 55% 99% 55% 98%
b2 IgA 55% 98% 31% 92%
b2 IgM 22% 96% 14% 97%
Conclusion: We have shown comparable clinical performance between
the BioPlex IgG and IgM assays and commercially available EIA kits.

In addition, testing on the BioPlex 2200 resulted in substantial reduc-
tion in the turnaround time compared to the conventional EIA.
Disclosure: Michael Watkins a Senior Staff Scientist at Bio-Rad
Laboratories, a major diagnostics manufacturer. Munther
Khamashta has nothing to disclose. Maria Bertolaccini has nothing
to disclose. Angela Tincani has nothing to disclose.
C115
How Many Tests Do We Need To Diagnose Antiphospholipid Syndrome
(APS)?
Cohen H1, Hills J1, Machin S1, Gardiner C1
1
University College London Hospitals, London, United Kingdom
The Sydney criteria for definite APS are useful for scientific clinical
studies. It has been suggested that the criteria could be simplified by the
omission of anticardiolipin (aCL), anti-b2glycoprotein-1 (ab2GP1) or
IgM antiphospholipid antibodies (aPL). However, diagnostic criteria
are required for routine clinical use. We analysed laboratory data from
a cohort of 193 patients with persistent aPL of whom 151 fulfilled
Sydney clinical criteria (thrombosis (arterial (n¼48), venous (n¼67))
and/or obstetric morbidity (n¼59); 23 had more than 1 manifestation).
Lupus anticoagulant (LA) testing was performed according to ISTH
guidelines. The Sydney laboratory criteria state that moderate/high
titre aCL are those above the 99th percentile or 40 GPLU/MPLU.
aCL positivity was defined as: low titre (95th percentile) IgG
>5GPLU/MPLU; moderate/high titre (99th percentile) >20 GPLU/
MPLU; ab2GP1 positivity was defined as: low titre (95th percentile)
IgG >3.5 units, IgM >3.0 units; moderate/high titre (99th percentile)
>15 units. In our experience, and that of others, the 99th percentile is
too insensitive for clinical use for women with pregnancy morbidity, so
the 95th percentile value was used in these patients. Of the 193 patients,
123 were considered to have APS (thrombosis with LA and/or mod-
erate or high titre aCL and/or ab2GP1; obstetric morbidity with LA
and/or aCL and/or ab2GP1). 53/123 patients with APS had LA. Of the
70 LA -ve patients, 12 had aCL only, 37 had ab2GP1 alone and 21 had
both. Of 94 patients who fulfilled clinical and laboratory Sydney cri-
teria, 41 were LA -ve, but had aCL and/or ab2GP1. 29/59 women with
a history of pregnancy morbidity and persistently weak þve aCL and/
or ab2GPI antibodies would have been classified as not having APS by
Sydney criteria. These data indicate that omission of testing for aCL or
ab2GP1 from the clinical investigation of APS could lead to a failure to
diagnose the syndrome in 9.7% and 30.0% of patients respectively,
and support the inclusion of low titre aCL and ab2GP1 in the diagnosis
of APS in women with pregnancy morbidity.
Disclosure: Nothing to disclose.
C116
Comparison of used and omitted tests in the recently updated guidelines
for lupus anticoagulant (LA) detection
Swadzba J1, Iwaniec T1, Pulka M1, Musial J1
1
Jagiellonian University Hospital, Cracow, Malopolska, Poland
LA is the most clinically relevant test between all antiphospholipid
antibodies. New guidelines for detection of LA were published
recently. The aim of the study was to compare methods suggested in
the newest guidelines with the other methods used for LA determina-
tion. The study group consisted of 336 subjects suffering from auto-
immune diseases. Thirty two suffered from PAPS; 80 from SAPS. The
remainder was the patients with SLE and SLE-like disease. One hun-
dred and forty one patients suffered from thrombosis (93 venous, 57
arterial) and 73 had pregnancy failures fulfilling APS criteria. LA
Abstracts
533
according to the updated guidelines was positive in 95 patients. Odds
ratio (OR) for thrombosis was 4,11 (95%: 2,5-6,8), for venous throm-
bosis was 4,01 and for arterial 2,03. The presence of LA was less
connected with pregnancy failures (OR¼ 1, 94, 95%: 1,1-3,7). When
only one test for LA would be consider, the sensitivity was lower but
specificity rather high with overall OR for thrombosis 5,04 and 4,34
respectively for aPTT and DVV. In the case of double positivity (based
on aPTT and DVV tests) OR substantially increased (6,47 for throm-
bosis, and 3,11 for pregnancy complications). Two other tests were
additionally used: a/ dPT and b/ the ratio between PTT reagent sensi-
tive and insensitive for LA. OR for thrombosis using dPT method was
higher (6,01, 95%: 3,6-9,9) than the ones when aPTT or DVV test were
used. Ratio between sensitive and insensitive aPTT was less clinically
relevant (OR for thrombosis: 3,54, 95%: 2,2-5,8). Finally presence of
LA dependent on b2glycoprotein-I was determined based on a method
described by P. de Groot. This test was the most clinically relevant (OR
¼ 8,36, 95%: 2,8-25,0 for thrombosis), while prothrombin-dependent
LA was clinically much less relevant for thrombosis (OR¼2,11, 95%:
1,2-3,8). The recently published guidelines for LA determinations give
easy to follow recommendations for standard laboratories, but they do
not include all the LA tests which may be clinically relevant.
Disclosure: Nothing to disclose.
C117
Dedicated to a complex diagnostic autoimmune diseases: the
anti-phospholipid antibody syndrome (APS)
Sorrentino A1, Rosita M1, Iervolino R1, Soriente I1, Vitagliano A2,
di Bella C3, Senatore RE4, Sabatini P1
1
umberto I hospital, nocera inferiore, salerno, Italy; 2Careggi Polyclinic
University, Firenze, Italy; 3Le Scotte Policlinic University, Siena, Italy;
4
tor Vergata University, Roma, Italy
The antiphospholipid syndrome (APS) is a systemic autoimmune dis-
ease associated with the presence of a heterogeneous group of antibo-
dies,difficult to diagnose early.The anti-cardiolipin antibodies (ACL)
and anti-b2glicoproteinaI (ab2GPI), are useful markers for the diagno-
sis and the follow up in APS. 134 patients aged 20 to 70 years, includ-
ing 63 women and 21 men with clinical suspicion of APS and 50
healthy patients, were subjected to the determination of three tests
criterion for the diagnosis of APS. Activated partial thromboplastin
time (aPTT), lupus anticoagulant (LAC) were performed on plasma,
while the dosage of anti-cardiolipin and anti-b2glicoproteinaI isotype
of IgG and IgM was performed with enzyme-linked immunosorbent
method assay (ELISA) using different production of diagnostic kits
and methods Fluorescence Enzyme-linked Immunosorbent assay
(FEIA). The ROC curves showed a different value of sensitivity and
specificity for assays of anti-cardiolipin and anti-b2glicoproteinaI car-
ried by ELISA.The FEIA method has a highly discriminating value
(p <0.0001) compared with assays performed in ELISA. The dosage of
biohumoral markers is essential for early diagnosis of APS and it’s
effective and efficient to take its course.
Disclosure: No response indicated.
C118
Validation of Bio-Rad anti-phospholipid and systemic autoimmune
assays for use on the Evolistm twin plus microplate system
Guo X1, Hardy J1, Sweet R1, Prestigiacomo T1
1
Bio-Rad Laboratories, Hercules, CA
Objective: Validate Bio-Rad Anti-phospholipid (aPL) and Systemic
Autoimmune (SA) assays for use on the EvolisTM Twin Plus (ETP)
Lupus
 Abstracts

534
fully automated ELISA system. Methods: The following Bio-Rad Table 2
ELISA assays were optimized and validated for use on the Evolis
Twin Plus system: they include Anti-Cardiolipin, Anti-Phosphatidyl Venous No venous
serine, and anti-Beta 2 Glycoprotein I; ANA Screening Test, Anti- Thrombosis thrombosis P-value when
(n ¼ 123) (n ¼ 191) age, sex, and
dsDNA, Anti-SS-A, Anti-SS-B, Anti-Sm, Anti-Sm/RNP, Anti-Scl-
Number Number race are
70, Anti-Jo-1, Anti-Centromere, and ENA Plus Screening. The three Assay (% positive) (% positive) P-value controlled
APL assays are offered in three Conjugates for the detection of IgA,
IgG and IgM, or IgM antibody detection, respectively. All assays are anti-beta2 IgA 29 (24 %) 32 (17%) 0.14 0.20
FDA cleared and distributed worldwide. The Evolis Twin Plus system anti-beta2 IgG 19 (15%) 8 (4%) 0.001 0.005
is a self-contained microplate processing system designed to completely aCL IgG 22 (18%) 13 (7%) 0.003 0.006
automate microplate-based EIA tests. It can handle assays that are aCL IgM 15 (12%) 19 (10%) 0.58 0.63
incubated at ambient temperature or elevated temperature such as anti-PSPT IgG 34 (28%) 23 (12%) 0.001 0.001
37oC. The ETP has a maximum capacity of three plates. Results: anti-PSPT IgM 27 (22%) 24 (13%) 0.041 0.13
The 95% confidence limits of slopes between manual and automated D4/5 IgA 20 (16%) 32 (17%) 1.00 0.94
runs were bounded by 0.8 to 1.2; R2>¼0.9. Total agreements for the
assays performed ranged from 85% to 100%. It was also demonstrated
that multiple assays can be combined in one given run. Conclusion: Conclusion: For any thrombosis, the anti-PS/PT assay performed the
This study has demonstrated that Bio-Rad Anti-phospholipid (aPL) best after adjustment for age, sex, and race. For venous thrombosis,
and Systemic Autoimmune (SA) assays product lines passed all valida- anti-beta2 GPI, aCL, and anti-PSPT IgG assays performed equiva-
tion criteria on the Evolis Twin Plus. The ability to combine multiple lently. For IgM assays, only the anti-PSPT was significantly associated
assays makes it possible to profile patient samples for multiple clinical with thrombosis. This study suggests that anti-PSPT may have value in
markers. Furthermore, the ability to automate these assays on the APS in SLE, and that only IgG assays may be cost-effective.
Evolis Twin Plus offers significant workflow savings for laboratories Disclosure: E. Akhter, None; W. L. Binder, INOVA Diagnostics Inc,
with ever increasing test volumes. Z. Shums, Inova Diagnostics, Inc., L. Magder, None; M. Petri, None.
Disclosure: Nothing to disclose.
C120
C119
Patient-derived monoclonal antibodies directed towards domain I of
Association of Traditional and Novel Antiphospholipid Antibody Assays b2-glycoprotein I display LAC activity
with Thrombosis in SLE Boon-Spijker M1, Dienava I1, de Groot PG2, Voorberg J1, Mertens K1,
Akhter E1, Binder W2, Shums Z2, Magder L3, Petri M1 erksen RHWM2, de Laat B2
1
1
Johns Hopkins University, Baltimore, MD; 2Inova Diagnostics Inc, San Sanquin Research, Amsterdam, Noord-Holland, The Netherlands;
2
Diego, CA; 3University Of Maryland, Baltimore, MD University Medical Center Utrecht, Utrecht, The Netherlands

Purpose: The current APS classification criteria recognize the LAC, Introduction: We have investigated the heterogeneity of the antibody
aCL and anti-b2 glycoprotein I. We determined the association of response against b2GPI by isolating human monoclonal antibodies
anti-PSPT and D4/5 IgA as well as anti-b2GPI and aCL, with throm- from B cells of two APS patients. Methods: B-cells were isolated
bosis in SLE. Method: Stored sera from 314 SLE patients in a pro- from two patients diagnosed with APS and cDNA coding for the
spective database were studied. 206 had a history of thrombosis variable heavy chain repertoire was cloned into pHEN1-VLrep.
and 108 did not. Assays were performed at INOVA Diagnostics, Inc. Single chain Fv fragments displayed on phages were selected for reac-
Results: Comparison of those with and without history of any type of tivity towards b2GPI. Clones were selected for full length IgG produc-
thrombosis with respect to the percent positive for each antibody. tion and tested for reactivity towards different domains of beta2GPI
and LAC activity. Results: phages were selected from the clones. Two
Four clones bound domain I (both VH4-4) and two clones bound
Table 1
b2GPI outside domain I (both VH3-23). Subsequently, we constructed
Thrombosis No thrombosis P-value when scFv’s from those 4 clones. Interestingly, for all four clones the trans-
(n¼206) (n¼108) age, sex formation from the phages (multiple binding sites) to scFv’s (one bind-
Number Number and race are ing site) resulted in an almost complete loss of reactivity for b2GPI. To
Assay (%positive) (% positive) P-value Controlled test whether this reduction in reactivity was based on the need for
bivalency, we constructed full length IgG’s. Indeed, all four clones
anti-beta2 IgA 45 (22%) 16(15%) 0.18 0.23 regained their reactivity towards b2GPI when produced as full-length
anti-beta2 IgG 24(12%) 3(3%) 0.010 0.021 IgG1. Both domain I-reactive monoclonal antibodies displayed LAC
aCL IgG 30(15%) 5(5%) 0.008 0.016 activity in an APTT-based clotting assay. Conclusion: We isolated
aCL IgM 27(13%) 7(6%) 0.086 0.11 human monoclonal antibodies directed against b2GPI from the Ig-
anti-PSPT IgG 47(23%) 10(9%) 0.003 0.001 repertoire of patients with APS. Functional characterization revealed
anti-PSPT IgM 43 (21%) 8(7%) 0.002 0.011 that domain I-specific antibodies display LAC activity. These human
D4/5 IgA 36(18%) 16(15%) 0.63 0.71 anti-domain I antibodies could be valuable tools (i) as a standard for
antiphospholipid antibody assays and (ii) to study the possible invol-
vement of these highly characterised antibodies in the clinical compli-
Table 2 shows comparison of those with and without a history of
cations observed in patients with APS.
venous thrombosis with respect to the percent positive for each
antibody. Disclosure: Nothing to disclose.

Lupus

C121
The 99th Percentile of aCL Are Lower in Japanese Population Compared
with Caucasian
Yamazaki M1, Nojima J1, Atsumi T1, Yasuda S1, Ieko M1
1 APhL
APS Group, SSC of Japanese Society of Thrombosis and Haemostasis,
Shinjuku, Tokyo, Japan
Purpose: According to revised classification criteria for APS, ‘‘medium
or high titer’’ of aCL corresponds to > 40 GPL (MPL) or > 99th
percentile of titers in healthy individuals. However, there is a possibi-
lity that the 99th percentile of aCL differs according to race, sex, and
age. Therefore, it is important to evaluate the titer in healthy indivi-
duals in consideration of those demographic factors. To achieve this
aim, we collaborated in the Asian Research Project entitled ‘‘Study of
regionality in laboratory test results and collaborative derivation of
reference intervals (RI)’’ and tried to set up cutoff value and RI for
aCL-IgG and –IgM in the Asian countries. Methods: The project was
conducted by participation of 3540 (2084 from Japan, 1456 from 7
Asian countries, between 20 to 64 years of age with even sex ratio)
healthcare professionals who had no remarkable past and present ill-
ness, no medication, and no remarkable abnormal laboratory data.
ACL-IgG and –IgM were measured by MESACUP Cardiolipin Test
IgG and IgM supplied by MBL Co., Japan. Results: By now, measure-
ment of 520 specimens from Japan was completed. IgG-aCL was not
influenced by region, sex, and age and its 99th percentile determined by
nonparametric method was 18.1 GPL, while 98% RI determined by
parametric method employing power transformation was 0.34 to 12.8
GPL. In contrast, IgM-aCL titer was higher in young female than in
male, and there is a trend of decrease with age in female. The 99th
percentile and the parametric 98% RI for aCL-IgM were 12.2 MPL
and 0.15 to 8.8 MPL in male, and 16.5 MPL and 0.25 to 12.9 MPL in
female, respectively. Conclusion: In contrast to the cut-off value pro-
posed by ISTH, the aCL titers in Japanese may be lower compared
with Caucasian. Age and sex-related changes were observed in IgM-
aCL titer, but not in IgG-aCL. We are planning to determine the 99th
percentiles and 98% reference interval for other aPLs such as ab2GPI
and aPS/PT from the same population.
Disclosure: Nothing to disclose.
C122
Predictive value and clinical significance of various antiphospholipid
(aPL) antibody tests in a multi-ethnic, multi-center SLE cohort
Seif A1, Aguilar-Valenzuela R1, Doan E1, Alarcon G3, Reveille J2,
Pierangeli S1
1
University of Texas Medical Branch, Galveston, TX; 2University of
Texas Houston, Houston, TX; 3University of Alabama at Birmingham,
Birmingham, AL
Objectives: To examine the predictive value (PPV), clinical significance
and association of single vs. multiple positivity with clinical manifesta-
tions of various aPL tests in a large multi-ethnic, multi-center SLE
cohort. Methods: Anticardiolipin (aCL) IgG, IgM, IgA, lupus antic-
oagulant (LAC), anti-b2glycoproteinI (anti-b2GPI) IgG, IgM, IgA
and a APhL ELISA (that utilizes a mixture of negatively charged
phospholipids instead of cardiolipin) were tested in serum/plasma sam-
ples of 899 SLE patients. Results: 559/899 SLE subjects were positive
for at least one of the aPL tests. 356 had thrombosis or pregnancy
losses.
Abstracts
535
Prevalence of positive aPL tests in SLE patients with APS manifestations
Tests Positives (%)
aCL 92
99
anti-b2 GPI 87
LAC 75
One test positive, others negative
aCL 5
APhL 6
anti-b2 GPI 4
LAC 3
When asked the question on how good each test run alone is in pre-
dicting thrombosis or pregnancy loss, the PPV were: APhL (96.2).
LAC (63.5), anti-b2GPI (60.7) and aCL (51.8). PPV was best when
all four tests were run compared to triple, double or single positivity.
Each of four tests was exclusively positive in 3-6 % of samples.
Conclusions: APhL test showed the best PPV when thrombosis and /
or pregnancy losses were considered. Importantly, the best PPV for
thrombosis and/or pregnancy loss was obtained when all four tests
were considered. The inclusion of the four aPL tests to confirm diag-
nosis of APS was needed to achieve the best PPV in this lupus cohort.
Disclosure: Silvia Pierangeli has financial interests with Louisville APL
Diagnostics, Inc.
C123
A new diagnostic strategy for early Anti-Phospholipid Syndrome (APS):
a gender oriented study
Iervolino R1, Clemente L1, Soriente I1, Sorrentino A1, Vitagliano A2,
Di Bella C3, Sabatini P1
1
Umberto I Hospital, Nocera Inferiore, Salerno, Italy; 2Medicine
University Firenze, Firenze, Italy; 3Medicine University Siena, Siena,
Italy
The gender is a social construction, which, unlike sex, bears on the
‘‘health outcome’’. The anti-phospholipid antibodies attack proteins
tying the phospholipids which are involved in the coagulation control,
like the b2glycoprotein1, the cardiolipin, the annexin V and the pro-
thrombin, causing thrombosis and the fetal loss. A cohort of 140
women aged 18 - 60 have been enlisted, of which 30 in fertile age
with clinical suspect of APS, 30 from the obstetrical division for abor-
tion or obstetric complications, 30 of which in menopause and 50
healthy donors. The blood plasma and serum of the selected groups
have been tested to: aPTT, lupus anticoagulant (LAC), the
d-dimers.the anti-cardiolipin and anti-b2glycoprotein1 IgG and IgM
isotype anti-bodies that has been done with the fluorescent-enzyme
immunoassay (FEIA), the anti-bodies anti-annexin V and anti-pro-
thrombin IgG and IgM isotype with the enzyme immunoassay
(EIA). The statistical analysis of the data has revealed a high extenu-
ating capacity (p< 0,0001) for the dosage of the anti-annexin V anti-
bodies and for the d-dimer test. Finally, the dosage of anti-
prothrombin isotype IgG antibodies, appears to be significant in
women affected by obstetric complications. The importance to use,
in the suspected or presence of APS, ‘‘diagnostic profiles divided by
gender’’ reduced the diagnostic error and it’s in harmony with the
modern concept of health.
Disclosure: No response indicated.
Lupus
 Abstracts

536
C124 ELISA. All results were compared to those of 171 apparently healthy
individuals. Results: Anti-prot Z antibodies were found in 26/119 SLE
Novel Antiphospholipid Antibodies and Stroke in SLE patients and 15/171 healthy individuals. Patients with SLE had anti-
Akhter E1, Binder W2, Shums Z2, Magder L3, Petri M1 prot Z more frequently than the healthy controls (OR 2.9 [95% CI 1.5-
1
Johns Hopkins University, Baltimore, MD; 2Inova Diagnostics, San 5.8], p¼0.002). However, levels of anti-prot Z were not different
Diego, Ca; 3University of Maryland, Baltimore, MD between patients with SLE and healthy controls (14.718.8 vs.
10.213.8, p¼0.9). No correlations were found between IgG anti-
Purpose: Stroke is the most common arterial thrombotic event in both prot Z and IgG aCL, anti-b2GPI, aPT or aPS-PT, or anti-protein S.
primary and secondary Antiphospholipid Syndrome (APS). We exam- The prevalence of anti-prot Z was not different between patients with
ined the association of several antiphospholipid antibodies and stroke thrombosis and those without (17.9% vs. 23.7%, OR 0.7 [95% CI 0.2-
in systemic lupus erythematosus (SLE). Methods: Stored sera from 68 1.8], p¼0.6). Levels of IgG anti-prot Z were also not different between
SLE patients with stroke and 246 SLE without stroke were studied. patients with thrombosis and those without (11.711.5 vs. 16.121.4,
D4/5 IgA is directed against domains 4/5 of beta2 glycoprotein I (anti- p¼0.2).
beta2). Assays for anti-beta2, anticardiolipin (aCL), anti-phosphatidyl- The prevalence and the levels of anti-prot Z are not different
serine-prothrombin (anti-PSPT) and D4/5 IgA were performed at
between patients with arterial or venous thrombosis and those without
INOVA Diagnostics, Inc. Results: Comparison of those with history
the event (16% vs. 23%, OR 0.6 [95% CI 0.2-2.0], p¼0.5 and 10.58.6
of stroke to those with no history of stroke with respect to the percent
vs. 15.820.6, p¼0.2 for the arterial thrombosis group, and 17.4% vs.
positive for each antibody is shown in
22.9%, OR 0.7 [95% CI 0.2-2.3], p¼0.8 and 12.113.0 vs. 15.319.9,
p¼0.5 for the venous thrombosis group). Conclusions: Although anti-
Stroke No stroke prot Z are frequently found in SLE, they do not appear to be strong
(n ¼ 68) (n ¼ 246) P-value when markers for thrombosis.
Number Number age, sex, and
(%) (%) race are Disclosure: Maria Laura Bertolaccini is funded by the Louise Gergel
Assay positive positive P-value controlled Fellowship.

anti-beta2 IgA 17 (25 %) 44 (18%) 0.22 0.21

anti-beta2 IgG 9 (13%) 18 (7%) 0.14 0.051 C126
anti-beta2 IgM 5 (7%) 17 (7%) 1.00 0.85
aCL IgA 4 (6 %) 6 (2%) 0.23 0.27 The value of anti-protein Z as a marker of adverse obstetric history in
aCL IgG 12 (18%) 23 (9%) 0.079 0.061 systemic lupus erythematosus
aCL IgM 9 (13%) 25 (10%) 0.51 0.56 Taib S1, Bertolaccini ML1, Murru V1, Hughes GRV1, Khamashta MA1
anti-PSPT IgG 17 (25%) 40 (16%) 0.11 0.010 1
Lupus Research Unit, The Rayne Institute, King’s College London
anti-PSPT IgM 13 (19%) 39 (15%) 0.46 0.23 School of Medicine, London, London, SE1 7EH, United Kingdom
D4/5 IgA 17 (25%) 35 (14%) 0.043 0.032
Background: Protein Z deficiency has been associated with adverse
obstetric history (AOH) and anti-prot Z have been found in women
Conclusion: Anti-PSPT has the most significant association with stroke with pathological pregnancies. We studied the prevalence and clinical
in SLE. Neither aCL nor anti-beta2 were significant. A novel IgA significance of anti-prot Z in a large cohort of SLE women. Patients
antiphospholipid antibody against domain 4/5 was also associated and Methods: Of 75 women (mean age 46.511; mean disease duration
with stroke. We conclude that anti-PSPT is the best aPL ELISA 13.58.8), 32 had an adverse obstetric history (22 with a history of
assay for stroke and that IgA D4/5 may also be useful. Further pro- miscarriages (<10th week of gestation) and a total number of 55 events;
spective studies are needed to confirm our findings. 12 with a history of fetal death (310th week of gestation) and a total
Disclosure: E. Akhter, None W. Binder, INOVA Diagnostics, Inc Z. number of 17 deaths; 6 patients with a total number of 10 premature
Shums, Inova Diagnostics, Inc. L. Magder, None; M. Petri, None. births at or before the 34th week of gestation). Only 20/32 fulfilled the
1999 Sapporo criteria for APS. Results: Anti-prot Z were present in
19% of the patients. The prevalence of anti-prot Z was not different
C125 between patients with AOH and those without (18.7% vs. 18.6%, OR
1.0 [95% CI 0.3-3.2], p¼1.0). Levels of anti-prot Z were not signifi-
The value of testing for anti-protein Z antibodies in systemic lupus cantly different between patients with AOH and those without
eythematosus (10.211.2 vs. 16.922.8, p¼0.1). For further analysis, patients with
Taib S1, Bertolaccini ML1, Hughes GRV1, Khamashta MA1 AOH were subdivided in different subgroups, according to the event
1
Lupus Research Unit, The Rayne Institute, King’s College London suffered. No differences were found in the prevalence and/or in the
School of Medicine, London, United Kingdom levels of anti-prot Z between patients with miscarriages and those with-
out (13.6% vs. 20.7%, OR 0.6 [95% CI 0.1-2.4], p¼0.7 and/or 9.28.7
Background: The significance of anti-prot Z in the development of vs. 16.121.6, p¼0.1). Although anti-prot Z were more frequently
thrombotic events is still not known. We analysed the prevalence found in patients with foetal death than in those without (33.3% vs.
and clinical significance of anti-prot Z in a large cohort of patients 15.8%), the difference did not reach statistical significance (OR 2.6
with SLE. Patients and Methods: We studied 119 SLE patients (116 [95% CI 0.6-10.5], p¼0.2). Levels of anti-prot Z were also not different
female, mean age of 41.712.3 years, mean disease duration 12.48.3 between these 2 groups (13.514.6 vs. 14.219.7, p¼0.9). No antibo-
years). Clinical records were carefully reviewed and patients were inter-
dies to protein Z were found in patients with history of prematurity.
viewed at the same time of sample collection. Nineteen patients had
Conclusions: Anti-prot Z antibodies do not appear to be a risk marker
APS and 40 had aCL and/or LA without fulfilling APS criteria. Thirty-
for adverse obstetric outcome.
nine patients gave a history of thrombosis. Among these patients, 16
had an arterial event, 14 had a venous event and 9 had had both Disclosure: Maria Laura Bertolaccini is funded by the Louise Gergel
arterial and venous events. Anti-prot Z were tested by an in-house Fellowship.

Lupus

C127
Experience of antiphospholipid antibodies in the real world: what is the
impact of a confirmatory test at 12 weeks according to Sapporo/Sydney
criteria?
Devignes J1, Wahl D1,2, Zuily S1,2, Lecompte T1,2
1
CHU, Vandoeuvre, France; 2UHP, Vandoeuvre, France
Background: Because of possible transient and epiphenomenal anti-
phospholipid antibodies, Sapporo/Sydney classification criteria for
antiphospholipid syndrome (APS) recommend a confirmatory test at
least 12 weeks apart from the first measurement. We have evaluated
the impact of these confirmatory tests in patients with a first positive
anticardiolipin (aCL) ELISA test. Materials, methods, results: ACL
IgG and IgM tests were performed as recommended and followed
the minimal requirements of the European Forum on antiphospholipid
antibodies. Among 6321 patients who were tested at least once and
included in our database we studied patients with medium or high
positivity who were further investigated. Mean age of patients was
47.5  14.6. Thirty-five percent were males, 65% females. We identified
91 patients with medium or high positivity of either IgG or aCL who
had subsequent confirmatory tests. The first confirmatory test was
performed with a mean interval of 83 days. Among patients with con-
firmatory tests after 12 weeks, 78.2 % were still present in medium or
high titre. A second confirmatory test was performed after a mean
interval of 227 days and 74.3 % of patients positive at the second
test still had medium or high titre positivity at the third test. Mean
values of aCL were: IgG 40.8 GPLU at first measurement and 55.7
GPLU at 3 months control and IgM 17.2 MPLU at first measurement
and 12.6 MPLU at 3 months control. Conclusions: These results indi-
cate that more than 20 % of patients with initial medium or high
positivity of aCL IgG or IgM will not meet the laboratory Sapporo/
Sydney classification criteria for APS after a 12 weeks interval. More
surprising additional testing at longer time intervals will further reduce
the proportion of patients meeting the criteria. This raises the question
of the optimal time frame for laboratory criteria of definite APS, at
least for aCL.
Disclosure: Nothing to disclose.
C128
The Obstetric Antiphospholipid Syndrome: from Sapporo to Sydney
update
S Voto LS1, Mattioli MJ1, Zarate R1, Pagliaro E1, Mainetti G1,
Fischer L1, Grand B1
1 patients and normal controls, while wide variations in antibody levels
Juan A Fernandez Hospital, CABA, Buenos Aires, Argentina
Introduction: Antiphospholipid antibodies are recognized as the most
frequent acquired risk factor for thrombophilia and as a treatable
cause of pregnancy complications. The clinical relevance of anticardio-
lipin antibodies (aCL) is actually an issue of debate and the role of
anti-beta-2-glicoprotein 1 b2GPI) in adding new patients with APS in
lupus anticoagulant (LA) and/or aCL negative patients is conflicting.
Objectives: To evaluate the diagnosis of APS according to Sapporo
Criteria compared to Sidney’s updated consensus in a group of
women with clinical/obstetric criteria of APS. Material and Methods:
187 patients (pts) with clinical/obstetric criteria of APS. Laboratory
tests: LA according to ISTH recommendations; aCL and b2GPI by
ELISA. Range values for Sapporo Criteria of aCL>20GPL/MPL;
b2GPI were not considered in the evaluation. Range values for
Sydney modifications of aCL>40GPL/MPL; anti-b2GPI >20U were
considered in the evaluation. Results: Pts were classified in two groups:
Sapporo Criteria (a) and Sydney update (b). a: APS was confirmed in
50 pts: 31 LA positive, 15 aCL> 20 and 4 LAþ aCL. b: APS was
Abstracts
537
confirmed in 47 pts: 34 LA positive; 2 aCL>40; 8 ab2-GPI >20 units
and 3 with combined laboratory positive test. Conclusions and
Comments: 1-The final number of pts with APS was similar in both
groups. 2-With determination of b2GPI >20U 8 new pts have APS, but
3/8 have aCL >20 and < 40U. 3-The modification of aCL cut off
excluded 11 patients that the only laboratory tests were aCL >20
and < 40U. Most of these pts have obstetric criteria of fetal death.
4- In spite of the advances, the recent updated consensus statement
does not contribute to the solution of the problems in diagnosing APS.
There is a need to improve the available laboratory criteria to define
the obstetric APS. 5-These aspects are important to define the thera-
peutic management in future pregnancies.
Disclosure: Nothing to disclose.
C129
Change in Antiphospholipid Antibodies Over Time in the COGNITION
Cohort
Brey R1, Holliday S2, Saklad A1, Zarzabal LA1, Petri M3, Reveille J4
1
UTHSCSA, San Antonio, TX; 2South Texas Veterans Health Care
System, San Antonio, TX; 3Johns Hopkins University, Baltimore, MD;
4
UT Houston HSC, Houston, TX
Objective: The purpose of this abstract is to describe the differences
between antibody levels and changes over time in patients with Lupus
as compared to a matched group of normal control subjects. Material
and Methods: Blood was obtained at each study visit (every 6 months)
and evaluated for levels of aCL IgG/M/A, anti-b2GP1 IgG/A, and
anti-ribosomal P (anti-P). A total of 187 Lupus patients and 181 con-
trols have been enrolled. Across all antibodies and subsequent visits
there were 407 samples available for analysis from SLE and 323 from
control subjects since the study onset. Results: For each antibody
tested (using Kruskall-wallis) the median values were significantly
higher in SLE as compared to control subjects (aCL IgG, IgA,
b2GP1 IgG and antiP all p<0.001; aCL IgM p¼0.003; b2GP1 IgA
p¼0.02). The frequency of positive values were seen (tested using
Fisher’s exact test) in Lupus versus control subjects as follows: aCL
IgG, 13.5% versus 1.3% (p<0.001); aCL IgM, 9.6% versus 3.9%
(p¼0.07); aCL IgA, 5.8% versus 1.3% (p¼0.06); b2GP1 IgG, 17.2%
versus 3.9% (p<0.001); b2GP1 IgA, 21% versus 11.8% (p¼0.03);
antiP, 11.5% versus 0 (p<0.001). Thirty-four subjects had samples
available for 5 consecutive visits. With the exception of 1 to 3 patients
for each antibody, there were no significant differences between initial
antibody value and values obtained over the course of the study period
(2 1/2 years). Conclusion: These findings suggest that in both Lupus
can occur over time, this appears to be unusual in this cohort. For the
majority of subjects, antibody levels remained stable over time.
Disclosure: This presentation was made possible by a grant from NIH/
NINDS.
C130
Do clinicallly relevant IgA anti-b2glycoprotein I (anti-b2 GPI) antibodies
bind to DIV/V of b2 GPI?
Martinez-Martinez LA1, Aguilar-Valenzuela R1, Seif A1, Binder W2,
Alarcon G3, Pierangeli S1
1
University of Texas Medical Branch, Galveston, TX; 2INOVA
Diagnostics, Inc., San Diego, CA; 3University of Alabama at
Birmingham, Birmingham, AL
Purpose: Published studies have shown that ‘‘pathogenic’’ IgG
anti-b2GPI mainly recognize epitopes in domain I of the protein.
Lupus
 Abstracts
538
Recently, it has been suggested that IgA anti-b2GPI antibodies may
recognize epitopes in domains IV/V of b2GPI and these antibodies
appear to be associated with certain manifestations of
Antiphospholipid Syndrome (APS). Objectives: a) to examine binding
specificity to domain IV/V of b2GPI of sera of patients for IgA anti-
b2GPI antibodies and b) to determine whether those antibodies are
associated with APS clinical manifestations. Methods: 80 IgA-anti-
b2GPI positive sera and data were selected from 588 SLE patients in
a multi-ethnic, multi-center cohort; Similarly, 35 IgA anti-b2GPI posi-
tive sera and data were obtained from 2188 patients that were referred
to our reference laboratory (APLS) for APS work-up. IgA anti-b2GPI
titers and binding of the sera to domain IV/V of b2GPI were examined
by ELISA using commercial kits (INOVA Diagnostics, Inc.) A positive
test was considered positive when values were above the corresponding
cut-off points. Results: A total of 62 out of 115 IgA (54%) anti-b2GPI
positive samples were positive in domain IV/V assay and 77 % of those
had clinical manifestations of APS that included: arterial and venous
thromboses, pregnancy losses, other APS-related pregnancy complica-
tions, seizures, thrombocytopenia, skin ulcers, pulmonary hyperten-
sion, livedo reticularis, cardiac valvular disease, and migraines.
Correlation of IgA anti-b2 GPI titers between the two kits was 0.69.
Conclusions: IgA anti-b2GPI antibodies that bind to domain IV/V of
b2GPI may represent an important subgroup of clinically-relevant aPL
antibodies.
Disclosure: Walter Binder has financial interests with INOVA
Diagnostics, Inc.
C131
Preanalytics of lupus anticoagulant (LA) testing: comparison of plasma
filtering, double centrifugation, and single centrifugation
Javela K1, Puurunen M1, Mustonen P1
1
Finnish Red Cross Blood Service, Helsinki, Finland
Background: Residual platelets in the plasma sample interfere with LA
testing. Platelets can be eliminated from plasma by filtration or efficient
centrifugation. Filtration has been shown to affect the level of some
coagulation factors, which could affect APTT-dependent coagulation
tests. According to NCCLS guidelines centrifugation time and dura-
tion must be established by the laboratory. Materials and Methods: In
our laboratory, about 4000 patient samples are tested for LA every
year. As screening, mixing test of dRVVT and PTT-LA (Diagnostica
Stago) are used in our laboratory. We have switched from plasma
filtration to double centrifugation (2 x 2500 g) three years ago and to
single centrifugation one year ago (1 x 2500 g). These three procedures
were compared. Forty samples representing different levels of LA (16
positive and 24 negative samples) were selected for comparison. The
results were presented as a ratio (clotting time of mixed patientþnor-
mal plasma/clotting time of normal plasma). Longitudinal patient
means are followed continuously as a part of our routine quality con-
trol. Results: Platelet count was <10 109/L in all samples. The repeat-
ability of LA screening tests was good (1.8-2.9 %) in both filtered and
double or single centrifuged plasmas. dRVVT results and long-term
patient means from filtered and double or single centrifuged plasmas
did not differ statistically significantly. However, PTT-LA levels were
significantly higher in filtered plasma samples (p<0.0001) than in cen-
trifuged samples. Long-term patient means of PTT-LA of filtered sam-
ples also were significantly higher. This has been taken into account
when setting new cut-off value. Conclusions: dRVVT screening test
results were similar in double and single centrifuged and filtered plas-
mas. However, results of double or single centrifuged plasmas tend to
be lower than in the filtered plasma. This favours centrifugation in LA-
pre-analytics. The result might reflect the effect of filtration on the level
Lupus
of coagulation factors. Single centrifugation of the sample was as good
as double centrifugation for LA testing.
Disclosure: Nothing to disclose.
C132
A serious problem in new lots of b2Glycoprotein I IgG assay
Javela K1, Puurunen M1, Mustonen P1
1
Finnish Red Cross Blood Service, Helsinki, Finland
Background: b2GP1 antibodies have recently been accepted as an offi-
cial laboratory criterion in terms of the updated classification criteria
for definite antiphospholipid syndrome (APS). A false positive result
could lead to serious consequences in patient care. b2GP1 IgG assay
has been performed in our laboratory since year 2000. About 4000
samples are analyzed yearly. The same reagent (QUANTALite b2
GPI IgG, INOVA, CA, USA) has been used all the time. This reagent
is widely used in many laboratories. The cut-off level is defined by our
own reference population. We have participated in ECAT Quality
Control surveys with good results since their beginning. Materials
and Methods: This paper reports results using five lots of the
QUANTA Lite b2 GPI IgG reagent during two years (lots 770098,
870092, 870144E6, 870359, and 970272). The same lot was in use for
approximately 6-12 months. These lots have been tested by our normal
protocol: 1) a routine check of the cut off level, as well as the level of
one positive and one negative control by using two commercial sam-
ples and one in-house control (near cut-off value; 20 SGU) and 2)
assessment of several patient samples with known b2GP1 levels (initi-
ally measured by the lot 770098). The same in-house control has been
used for 2-3 years and is never changed simultaneously with a reagent
lot. Results: We detected significant changes in patient and control
samples and patient means when testing the lots 870092, 870144E6,
and 870359: the results were two times higher, 40% lower, and 30%
lower, respectively, than by the established reference lot (770098). The
results had to be divided by 2, multiplied by 1.7, or multiplied by 1.4,
respectively, to get the same results as by lot 770098. At last, by the lot
970272 we got same results as by lot 770098. Conclusions: Several
reagent lots (n¼10) have earlier been used in our laboratory without
any problems. The last three lots, however, have turned out to be
unreliable. The manufacturer has paid some attention to the fluctua-
tion between the lots, and finally the level of results has returned to the
original level. When the difference was highest, the possibility of a false
APS diagnosis would have been ten times higher if we had not noticed
the marked level change of results.
Disclosure: Nothing to disclose.
C133
Are IgA anti-b2GPI antibodies clinically relevant?
Seif A1, Alarcón G4, Aguilar-Valenzuela R1, González E1, Binder W3,
Theodorescu M2, Pierangeli SS1
1
University of Texas Medical Branch, Galveston, TX; 2Theratest
Laboratories, Lombard, IL; 3INOVA Diagnostics, Inc., San Diego, CA;
4
University of Alabama at Birmingham, Birmingham, AL
Purpose: The clinical significance of IgA aPL antibodies is uncertain.
Objectives: a) to examine the prevalence of exclusive IgA-anti-b2GPI
antibody positivity in a large cohort of patients with SLE and in
patients suspected of having APS; b) to correlate IgA anti-b2GPI
positivity with APS-associated clinical manifestations. Methods: aPL
seropositivity was examined in sera of 588 SLE patients from a multi-
ethnic, multi-center cohort (LUMINA) and in 2108 sera from patients
that were referred to our laboratory (APLS) for APS work-up between

Jan-2008 and May-2009 by ELISA. IgA anti-b2GPI titers were deter-
mined in two commercial ELISA. Results: Thirty-six patients’ samples
from the LUMINA cohort and 23 from the APLS laboratory were
positive exclusively for IgA-anti-b2GPI. All the samples were positive
for IgA-anti-b2GPI antibodies in at least one kit. Sixty-nine % of the
LUMINA subjects and 78% in the APLS group had at least one APS-
related clinical manifestation that included: arterial and venous throm-
boses, pregnancy losses, other APS-related pregnancy complications,
seizures, thrombocytopenia, skin ulcers, pulmonary hypertension,
livedo reticularis, cardiac valvular disease, and migraines. One patient
in each group was LAC positive. Conclusions: This study supports the
notion that elevated IgA anti-b2GPI antibody titers may identify addi-
tional patients who have clinical features of APS but who do not meet
current diagnostic criteria. It may be therefore recommended to test for
IgA anti-b2GPI antibodies when other aPL tests are negative and APS
is suspected.
Disclosure: Teodorescu M and Binder W have commercial interests
with Theratest Labs and INOVA diagnostics, Inc.
C134
Clinical significance of IgG and IgM autoantibodies that target the
complex of phosphatidylserine and prothrombin (PS/PT)
Binder W1, Lewis S1, Shums Z1
1
INOVA Diagnostics, Inc., San Diego, CA
Objective: To demonstrate performance characteristics and clinical uti-
lity of an ELISA assay for detecting autoantibodies that react with a
complex of phosphatidylserine and prothrombin (PS/PT). Method: We
tested 71 patients with anti-phospholipid syndrome (APS), 24 known
Lupus Anticoagulant (LAC) positives, 247 random normals and 52
disease controls for IgG and IgM antibodies to PS/PT. These results
were used to calculate performance characteristics and the new assays
were compared to traditional anti-b2GPI and LAC assays. Matched
serum and citrated plasma samples were tested to see if sample matrix
had any influence on results. Results: All LAC positive patients were
found to be strongly positive for either IgG, IgM PS/PT antibodies or
both. Most patients with APS were found to be PS/PT positive. Forty
eight of the 71 APS patients (67.6%) were PS/PT positive and many of
these individuals were found to be negative using more traditional
assays such as anti-b2GPI and LAC. Only 7 of 247 normals and 1 of
the 52 disease controls were found to be positive for either IgG or IgM
PS/PT antibodies for a combined specificity of 97.3% (8/299). The
assays were found to have high inter and intra run precision.
Equivalent results were obtained with either serum or citrated
plasma. Conclusion: IgG and IgM autoantibodies that react with a
physiologic complex of phosphatidylserine and prothrombin are sensi-
tive markers for anti-phospholipid syndrome. These antibodies are
present in APS patients that are negative by methods currently in
use. The tests exhibit high specificity and reproducibility and can be
run with serum or plasma specimens.
Disclosure: Walter Binder, PhD, Steve Lewis and Zakera Shums are
employees of INOVA Diagnostics, Inc.
C135
Diagnostic Performance of Anti-Phosphatidylserine-prothrombin
Complex (aPS/PT) Antibody Assays in the Evaluation of
Antiphospholipid Syndrome and Recurrent Pregnancy Loss
Jaskowski TD1, Wilson A1, Hill HR1,2, Branch WD2, Tebo AE1,2
1 aPT were not detected.
ARUP Laboratories, Salt Lake City, UT; 2University of Utah, Salt
Lake City, UT
Abstracts
539
Determination of anti-phosphatidyl serine/prothrombin complex (aPS/
PT) antibodies has been suggested to be of alternate diagnostic value to
lupus anticoagulant (LA) activity in the evaluation of antiphospholipid
syndrome (APS). To determine their clinical performance and associa-
tion with other antiphospholipid (aPL) antibodies, we measured aPS/
PT (two manufacturers), anti-phosphatidyl serine (aPS) and anti-pro-
thrombin (aPT) IgG and IgM antibodies by ELISA. All samples were
also tested for anti-cardiolipin (aCL) and anti-beta 2 glycoprotein I
(ab2GPI) IgG and IgM antibodies as well as lupus anticoagulant (LA)
unless otherwise indicated. Three groups were studied: (i) APS with
recurrent pregnancy loss (RPL) and/or thrombosis (n ¼ 62); (ii) RPL
only (n ¼ 66) and (iii) healthy women with successful pregnancies
(WSP; n¼30). Assays predictive capabilities were determined by recei-
ver operator characteristic (ROC) curves and area under the curve
(AUC) analyses, concordance was evaluated by Cohen’s Kappa (k)
statistics. No statistically significant difference between the predictive
powers of the two aPS/PT assays was observed. There were also no
significant differences between both aPS/PT and aPS assays. However,
significant differences between the predictive power of the aPS/PT and
aPT assays (p¼0.002) were demonstrated. The agreement between the
aPS/PT antibody tests and LA for the APS cohort was very poor
(k<0.2). Overall, our study shows good agreement between the two
commercial assays for the detection of aPS/PT antibodies. However,
the poor association between these two assays and the LA test pre-
cludes the replacement of the LA in the initial evaluation of APS.
Disclosure: Nothing to disclose.
C136
Only aPS/PT ELISA detects low avidity antiprothrombin antibodies
Zigon P1, Ambrozic A1, Cucnik S1, Bozic B1,2, Kveder T1, Rozman B1
1
University Medical Centre, Ljubljana, Slovenia; 2University of
Ljubljana, Ljubljana, Slovenia
Objectives: By different ELISAs two types of antiprothrombin antibo-
dies (aPT) are detected: one recognizing prothrombin bound to PS
(aPS/PT) and the other prothrombin alone (aPT-A). No data on avid-
ity of aPT has been published however, they may be of different avidity
which could explain many inconsistencies in their detection. The aim of
our study was to determine aPS/PT, aPT-A and their avidity. Methods:
aPS/PT and aPT-A were measured in sera of 194 patients: 95 APS, 37
SLE, 50 RA and 12 SjS. For avidity testing a chaotropic aPS/PT
ELISA with increased concentrations of NaCl in the binding buffer
was used. High avidity aPT: the binding of antibodies at 0.5M NaCl
remained higher than 70% of the initial binding at 0.14M NaCl. Low
avidity aPT: the binding decreased below 30%. The remaining were
declared of heterogeneous avidity. Results: 44/95 APS and 5/37 SLE
patients were aPS/PT positive while 14/95 APS and 3/37 SLE patients
tested aPT-A positive. RA and SjS patients were all negative.
Specificity of ELISAs was comparable (80% vs. 83%) but sensitivity
of aPS/PT ELISA was higher than of aPT-A ELISA (60% vs. 25%)
Chaotropic aPS/PT ELISA was selected for avidity determination.
aPT were of low, heterogeneous and high avidity in 6,35,8/44, respec-
tively. Most important none of the low avidity, 26% of heterogeneous
and 75% of high avidity antibodies were positive in aPT-A ELISA.
Among patients having heterogeneous or high avidity aPT significantly
more had APS than among low avidity (40/43 vs.4/6; p<0.05). None of
SLE patients had high avidity aPT. Conclusion: Both versions of
ELISAs detected heterogeneous and high avidity aPT while low avidity
were only detected with our in-house aPS/PT ELISA. APS patients had
aPT of very heterogeneous avidity while in SLE patients high avidity
Disclosure: Nothing to disclose.
Lupus
 Abstracts
540
C137
Clinical Utility of a Novel Screening Assay for the Detection of IgG and
IgM Antibodies to the Phosphatidylserine/Prothrombin Complex
Shums Z1, Encabo S1, Van M1, Norman G1, Binder W1
1
Inova Diagnostics, San Diego, CA
Objective: To evaluate the performance and clinical utility of a proto-
type anti-phosphatidylserine/prothrombin (PS/PT) screening ELISA
designed to detect both IgG and IgM antibodies to PS/PT complex
in human serum or plasma. Method: Sera from 16 patients with a
known clinical diagnosis of APS, 88 sera from random healthy con-
trols, and 24 sera known to be Lupus Anticoagulant (LAC) positive
were tested on the new anti-PS/PT IgG/IgM ELISA assay (INOVA
Diagnostics, San Diego, CA). Sera were also tested on individual PS/
PT IgG and PS/PT IgM ELISA kits (INOVA Diagnostics, San Diego,
CA). Results: While neither the IgG nor the IgM PS/PT ELISAs were
sufficient individually to detect all the LAC positive sera, testing of the
LAC sera with both IgG and IgM PS/PT assays resulted in detection of
all 24 LAC sera. By testing for both IgG and IgM simultaneously, the
new PS/PT IgG/IgM ELISA assay was able to efficiently detect all 24
LAC positive patients, showing a 100% correlation with the conven-
tional LAC test results. Twelve of the 16 sera (75%) from APS patients
were positive by the PS/PT IgG/IgM ELISA. Some of these APS sera
were previously found to be negative by conventional assays such as
anti-b2GPI and LAC. Of the 88 healthy controls tested, one patient
had a borderline positive unit value of 31 units (cutoff is 30 units)
giving a specificity of 98.9% (87/88). Conclusion: As a result of the
combined detection of both IgG and IgM PS/PT antibodies, the pro-
totype anti- PS/PT screening assay was able to detect 100% (24/24) of
the LAC sera tested. In addition it detected 75% of the APS sera tested
with very high specificity. This assay may prove to be a very valuable
tool for the screening of patients with suspected anti-phospholipid
syndrome and it may be useful in screening for LAC positives.
Disclosure: All the authors on the abstract are employed at Inova
Diagnostics.
C138
Two new assays, anti-prothrombin IgG and anti-prothrombin IgM, are
automated on the PHDTM and the next generation PHDTM lx systems
Guo X1, Sweet R1, Hardy J1, Atkinson D1, Cox A1, Prestigiacomo T1
1
Bio-Rad Laborartories, Hercules, CA
Objectives: Automation of the Bio-Rad EIA Anti-Prothrombin IgG
and Anti-Prothrombin IgM Tests. Methods: Antibodies to prothrom-
bin have been identified in patients with antiphospholipid syndrome
(APS). Clinically, elevated serum levels of these antibodies are asso-
ciated with an increased risk for APS, characterized by recurrent arter-
ial or venous thrombosis, thrombocytopenia, and/or fetal loss. APS
may be found in patients with autoimmune diseases (e.g., SLE or
secondary APS) as well as in individuals without an obvious under-
lying disease (primary APS). Two tests, Bio-Rad EIA Anti-
Prothrombin IgG and Anti-Prothrombin IgM, both FDA approved
for marketing, are evaluated for their performance with automation on
the PhDTM Microplate System and the PhDTM lx Microplate Systems.
Both instruments are fully automated microplate processing instru-
ments, capable of sample dilution, sample dispense, incubation, wash-
ing, reagent dispense and data reduction. This study demonstrates the
performance of the Bio-Rad EIA Anti-Prothrombin IgG and Anti-
Prothrombin IgM Tests. Results: Data will show the performance of
the assay manually and with automation. Performance
characteristics for precision, within-assay drift, and correlation
between the different assay formats are presented. Conclusions: The
Lupus
Bio-Rad Anti-Prothrombin IgG and Anti-Prothrombin IgM Tests
have passed all validation study acceptance criteria for running on
the PhD and the PhD lx microplate systems and provide equivalent
performance to the manual assays. The fully automated Anti-
Prothrombin IgG and Anti-Prothrombin IgM tests on both systems
will provide consistent and reliable information in the aid of clinical
diagnosis of primary and secondary APS.
Disclosure: Nothing to disclose.
C139
Prevalence of antibodies to prothrombin (PT) and prothrombin/
phosphatidylserine (PS) in a cohort of lupus anticoagulant (LAC)
positive samples
Aguilar-Valenzuela R1, Schleh A1, Khan U1, Belter M2, Doan E1,
Gould N2, Dlott J2, Pierangeli S1
1
University of Texas Medical Branch, Galveston, TX;
2
Quest Diagnostics, Chantilly, VA
Background: ELISA tests have been developed to detect anti-PT and
anti-PT/PS antibodies in serum but the clinical diagnostic value and
their relationship with the LAC remains elusive. Objective: to evaluate
anti-PT/PS and anti-PT antibodies in a cohort of 150 confirmed LAC
and the corresponding controls. Methods: 150 LAC positive and 150
LAC negative plasma samples were tested by ELISA for anti-PS/PT
(IgG and IgM) and LAC PS/PT screening assay (INOVA Diagnostics)
and anti-PT (IgG and IgM) in-house assay. LAC positivity was con-
firmed by either dRVVT Confirm (American Diagnostica) or by the
STACLOT LA (Diagnostica Stago) tests (3% of samples were positive
for dRVVT only and 94% were positive for STACLOT only, 3% were
positive for both). Results:
aPS/PT aPS/PT aPS/PT aPT aPT
IgG IgM Screen IgG IgM
# of positive 43/150 33/150 32/150 44/150 39/150
samples (%) (29%) (22%) (21%) (29.3%) (26%)
All negative LAC samples were negative in the anti-PS/PT and anti-PT
tests. 125/150 (83%) of all LAC positive samples were positive in at
least one of the PS/PT or PT ELISA. There was no correlation between
any of the tests and LAC Staclot delta values or dRVVT ratios and the
PT/PS or PT tests. Strong correlation was observed between anti-PS/
PT IgG or IgM and LAC Anti-PS/PT screen test. Conclusions: PS/PT
and PT antibodies altogether recognize the majority of LAC positive
samples and may represent an additional specific serological laboratory
marker to confirm diagnosis of Antiphospholipid Syndrome.
Disclosure: Nothing to disclose.
C140
D-Dimers reference values determination in pregnant normal women
through ELISA methodology
Otero A1, Lena A1,2, Attarian D1, Mota N1, Dominguez G1, De Los
Santos S1, Pita D1
1
CEAHT, Montevideo, Uruguay; 2HCFFAA, Montevideo, Uruguay
Introduction: D-Dimers evaluation by ELISA methodology is very
useful for diagnostic and follow-up of pregnant women in risk of preg-
nancy failure. It is considered a way of controlling the effectiveness of the
therapy in patients treated with low molecular weight heparin. The aim
of this work is to determine D-Dimers reference values in pregnant

normal women by the mentioned methodology. Methods: Pregnant
normal women were selected in the first (n¼50), second (n¼99) and
third (n¼96) trimesters who might have had at least one previous preg-
nancy without complications. D-Dimers were determined by means of
ELISA methodology to each one of these patients. Statistical calcula-
tions were made in order to determine reference values. Results:
D-Dimers distribution values for second and third quarter resulted
to be parametric, so media and standard deviations were estimated
for them. Distribution values for the first one resulted not parametric
so median and 2.5 and 97.5 percentiles were estimated, resulting the
following: First quarter Median 400 ng/ml Reference ranges 193-
662ng/ml; Second quarter Media 836ng/ml Reference ranges 262-
1410 ng/ml; Third quarter Media 1153 ng/ml Reference ranges
285-2021 ng/ml. Conclusions: It is known that D-Dimers are expected
to be less than 400 ng/ml in normal notn-pregnant individuals. We
concluded that D-Dimers suffer physiological variations during preg-
nancy: an increase which begins in the first quarter and continues
during the whole pregnancy takes place in normal pregnant women.
It is important to have established these reference values to compare
them to the results obtained in patients upon treatment, which will be
the aim of a future study.
Disclosure: This presentation was made possible by a partial support
by ROCHE DIAGNOSTICS.
C141
Plasma levels of platelet-derived microparticle (PDMP) may be an useful
indicator of antiplatelet therapy in antiphospholipid syndrome (APS)
Yamazaki M
Kanazawa University Graduate School of Medicine, Kanazawa,
Ishikawa, Japan
Background: Adequate indicators for antiplatelet therapy are few.
Recently, plasma levels of platelet-derived microparticles (PDMP),
which released from activated platelets and whose increment may
reflect platelet activation in vivo, could be measured using ELISA.
To estimate whether plasma PDMP level is a useful indicator of anti-
platelet therapy, we treated 60 patients in antiphospholipid syndrome
(APS) with several kinds of antithrombotic therapies and evaluated for
plasma PDMP level and clinical features. Methods: Sixty APS patients
with previous stroke were randomly treated with 100 mg of aspirin
alone, aspirin plus cirostazol, an antiplatelet agent, at dose of 100
mg twice a day or with aspirin plus warfarin for 3 years. In aspirin
plus warfarin group, PT-INR was adjusted 2.0 to 2.5. Brain MRI
studies were performed every 6 months. Plasma levels of PDMP and
prothrombin fragment (F1þ2) were measured every 3 months as well
as routine blood examinations. We received informed consent from all
study subjects before treatment. Results: Mean plasma PDMP levels
significantly increased in APS patients compared with those in normal
subjects (17.6  3.9 ng/mL vs. 10.5  2.5 ng/mL, p < 0.01). Those
decreased in aspirin plus cirostazol group and in asnpirin plus warfarin
group (p < 0.01 and < 0.05, respectively), but not in aspirin group.
MRI examinations revealed that recurrences of stroke were shown in 3
APS patients treated with aspirin alone in 1 year follow-up and 2 with
aspirin plus warfarin in 3 year-period. Interestingly, all of those 5 APS
patients with recurrent stroke showed elevation in plasma PDMP levels
but not F1þ2 levels before its recurrence. Conclusions: These observa-
tions suggest that Plasma PDMP level may be useful for an indicator
of antithrombotic therapy in APS patients. That may be applicable for
adoption of antithrombotic therapy and therapeutic indicator in indi-
vidual cases.
Disclosure: Nothing to disclose.
Abstracts
541
C142
Combination therapy of antiplatelet agents may be effective for
prevention against arterial thromboses in antiphospholipid syndrome
Yamazaki M
Kanazawa University Graduate School of Medicine, Kanazawa,
Ishikawa, Japan
Background: Previous prospective study revealed that intensive antic-
oagulation with warfarin has preventive effect for arterial thromboses
in antiphospholipid syndrome (APS). However, those effects of
aspirin-based combination therapies with other antiplatelet agents or
with low dose warfarin have not been established yet. Purpose: To
clarify whether those combination therapies have protective effect
against arterial thromboses in APS patients, we designed a prospective
study. Sixty APS patients with previous stroke were randomly treated
with 100 mg of aspirin alone, aspirin plus cirostazol, an antiplatelet
agent, at dose of 100mg twice a day or with aspirin plus warfarin for 3
years. In aspirin plus warfarin group, PT-INR was adjusted 2.0 to 2.5.
Brain MRI studies were performed every 6 months. We received
informed consent from all study subjects before treatment. There
were no significant changes in background diseases and in general
risk factors among 3 groups. Results: After 1 year-period, 3 of 20
patients with aspirin alone but none with combination therapies
showed new small infarctions on MRI study. Therefore, the aspirin
single treatment group discontinued it for humanitarian considera-
tions. None of patients treated with combined antiplatelet agents
showed new stroke, and 2 of 20 with aspirin plus warfarin showed
recurrent strokes on brain MRI by 3 years-period. No critical adverse
effects had been shown in any patients. Conclusion: These results sug-
gested that aspirin-based combination therapies, especially, combined
antiplatelet therapy may be effective for prevention of arterial throm-
boses in APS patients. Randomized large prospective study is needed
for establish of protective effect of it.
Disclosure: Nothing to disclose.
C143
Pregnancy outcomes in women with acquired or inherited thrombophilias
treated with aspirin þ enoxaparin
Gerde M1, Heer C1, Landeira A1, Carames C1, Ballarin M1, Boechat F1,
Martin N1, Balparda J1
1
Austral University Hospital, Pilar, Buenos Aires, Argentina
Background: Thrombophilias are associated with a higher risk of
adverse pregnancy outcomes or thrombotic events. Anticoagulant
therapy with heparin seems to improve these results. Methods: This
is a retrospective evaluation of the effectiveness and safety of throm-
boprophylaxis in pregnant women between 25-42 years old with pri-
mary or secondary antiphospholipid syndrome or with Antihrombin
III deficiency, Protein C or S deficiency, FV Leiden mutation,
Protrombin G20210A mutation. All of them had history of pregnancy
loss, preterm delivery < 34 weeks due to severe preeclampsia or pla-
cental insufficiency or history of thromboembolism. Aspirin (125mg þ
Vitamin C) þ 40 mg enoxaparin daily (or 80mg if previous thrombosis)
was given since conception was attempted until four weeks postpar-
tum. Results: 127 women fulfilled inclusion criteria and there were 137
pregnancies. 65 patients had primary and 6 secondary antiphospholi-
pid syndrome, 27 hereditary thrombophilia and 29 two or more throm-
bophilic defects. 89% (122/137) pregnancies resulted in live births. 14
(10%) were embryonic losses (< 10 weeks) and only 1/137(1%), a
second trimester loss (14 weeks). There was no fetal death > 14
weeks. From 122 live births, 94 % were term deliveries and average
weight was 3200g. There was no VTE during pregnancy or
Lupus
 Abstracts
542
puerperium.1 patient had excessive bleeding during cesarean section.
Conclusion: Treatment with AAS þ enoxaparin proved to be effective
and safe in preventing adverse obstetric outcomes and VTE in preg-
nant women with thrombophilia.
Disclosure: Nothing to disclose.
C144
The Effects of Hydroxychloroquine on Antiphospholipid Antibody Titers
in SLE
Broder A1, Putterman C1
1
Albert Einstein College of Medicine, Bronx, NY
Background: Hydroxychloroquine (HCQ) therapy is associated with a
decreased risk of thrombosis. In vitro, HCQ decreases the binding of
aPL-beta2 GPI complexes to phospholipid bilayers. We investigated
the changes in aPL titers over time vs. HCQ therapy in SLE. Method:
We included SLE patients with aPL or LAC measured at least twice
between 2002 and 2009. At baseline, patients were defined as
aPL/LAC- if they tested negative for both aPL and LAC, and as
aPL/Lacþ if they tested positive for either aPL or LAC, using labora-
tory cut-offs. An ‘‘event’’ was defined as aPL and/or LAC conversion
from positive to negative in aPL/LACþ, and as conversion from nega-
tive to positive in the aPL/LAC. ‘‘Time to event’’ was defined as
months from baseline to censoring or conversion. Patients were defined
as HCQþ/HCQ- based on HCQ treatment status at the event time.
Eighty four patients met the inclusion criteria: 31 HCQ-, 27
HCQþ, and 26 missing HCQ data. HCQþ and HCQ- groups were
similar in terms of demographic and disease related parameters at the
event time. During the course of follow-up, HCQþ patients were more
likely to be prescribed prednisone (p¼0.004) and/or DMARDs includ-
ing cyclophosphamide, azothioprine, or mycophenolate mofetil
(p¼0.02). Twenty (64%) patients in the HCQ- group and 9 (33%) in
the HCQþ group (p¼0.02) were placed on warfarin during the course
of follow-up, suggesting that there was a higher rate of thrombotic
events in the HCQ- group. Thirty two patients, 16 (50%) HCQþ
and 16 (50%) HCQ-, were aAPL/Lac- at baseline. Three (9%) patients
became aPL/Lacþ. All 3 were in the HCQ- group. The differences were
statistically significant by the log-rank test (p¼0.0026). Twenty six
patients, 11 (42%) HCQþ and 15 (58%) HCQ-, had at least one
aPL/Lacþ at baseline. Six (55%) were on HCQ and 3 (20%) were
not on HCQ at the time of conversion to all aPL/Lac-. The differences
were statistically significant by the log-rank test (p¼0.0009). The mean
decrease of aPL levels was 10.9 GPL in HCQ- group and 20.25 GPL in
the HCQþ group. Conclusion: Our data indicate that HCQ may play a
role in decreasing APL antibody levels in aPLþ patients and maintain-
ing low aPL levels in aPL- patients. This observation is consistent with
previously published laboratory data. Long-term prospective studies in
a larger group of patients are needed to confirm this observation. If
confirmed, HCQ may be used to treat APLS in the future.
Disclosure: Nothing to disclose.
C145
Simvastatin can prevent the procoagulant/thrombogenic properties of
antiphospholipid antibodies in monocytes
Lambrianides A1, Bell K1, Heywood W1, Mills K1, Isenberg D1,
Latchman D1, Pierangeli S2, Rahman A1, Giles I1
1
University College London, London, United Kingdom; 2University of
Texas, Galveston, TX
A major mechanism of hypercoagulability in the antiphospholipid
syndrome (APS) is antiphospholipid antibody (aPL)-mediated
Lupus
up-regulation of tissue factor (TF) on monocytes. Statins have been
shown to modify the function of endothelial cells and platelets by
decreasing the expression of adhesion molecules, inhibiting TF expres-
sion and down-regulating inflammatory cytokines after treatment with
aPL. The aim of this study was to determine whether simvastatin has
an effect on the APS-IgG-mediated up-regulation of p38MAPK and
NFkB signalling pathways and TF activity in monocytes. Furthermore
using proteomic analysis, our aim was to identify other proteins that
may be altered. IgG was purified from 5 APS patients with vascular
thrombosis (VT) and 5 healthy controls. A human monocyte cell line
was treated with 100mg/ml IgG for 6 hours. In some experiments, cells
were pretreated with simvastatin (5mM). The cell extracts were exam-
ined by immunoblot using total and phospho-specific antibodies to
p38MAPK and NFkB p65 and by fluorescence 2D difference gel elec-
trophoresis (DIGE). TF activity was also measured. IgG from patients
with VT increased the up-regulation of p38MAPK, NFkB and TF
activity in monocytes and this up-regulation is inhibited in the presence
of simvastatin. We have identified that there are 24 proteins showing
significant differences in expression in monocytes treated with IgG
from patients with VT in the presence of simvastatin – the majority
with decreased expression between 2.1 and 5.2 fold. Our findings
demonstrate, for the first time, that statins inhibit the APS-IgG-
mediated up-regulation of p38MAPK, NFkB and TF activity and sig-
nificantly altered the expression/modification of proteins in monocytes.
This supports the possibility that statins may offer an alternative non-
anticoagulant therapeutic approach for treating APS.
Disclosure: Nothing to disclose.
C146
Antiphospholipid syndrome and management of pregnancy complications
Makatsaria A1, Bitsadze V1, Baimuradova S1, Akinshina S1,
Khizroeva J1
1
Moscow Sechenov Medical Academy, Moscow, Russian Federation
Aims: To determine thrombophilia and to evaluate maternal and fetal
outcomes in women with history of pregnancy complications receiving
the preconception treatment. Material and Methods: 400 pts with fetal
loss syndrome, 160 pts with severe preeclampsia, 80 pts with VTE, 100
pts with placental abruption, 60 pts with antenatal death and 500
healthy controls were tested for thrombophilia and APA. Women
with history of pregnancy complications received treatment in the pre-
conception period and during pregnancy. Results: Thrombophilia and
APA were found in 82% women with fetal loss syndrome, in 96,4%
women with recurrent preeclampsia and in 100% women with placen-
tal abruption and VTE. In women with fetal loss syndrome MTHFR
C667T, APA, FV Leiden, prothrombin G20210A, multigenic throm-
bophilia were found in 41.3%, 29.3%, 15.3%, 4% and 20% respec-
tively, in women with preeclampsia - in 44.6%, 16%, 17.8%, 3.6%,
73.2%, in women with placental abruption and VTE in 71%, 25.3%,
37.4% and 96.4%, in controls in 12%, 4%, 2%, 1.3% and 4% cases
respectively. Prevalence of thrombophilias and APA in study group
was higher vs controls (p<0,05). APA spectrum in women with fetal
loss syndrome: LA - 16.6%, aCL -8.8%, anti-beta2-GPI – 7,3%, anti-
annexin V – 17.7%, anti-prothrombin – 1.9%. Preconception therapy
allowed preventing recurrent fetal loss syndrome in 66%; 96% patients
were delivered after 37 weeks. In the study group nobody had moder-
ate or severe form of preeclampsia, mild preeclampsia was observed in
16%, all babies were alive. Patients had not recurrence of placental
abruption or VTE, all babies were alive. Conclusions: Thrombophilia
might be the main pathogenetic mechanism of recurrent pregnancy
complications. Due to thrombophilia involvement in trophoblast inva-
sion and placentation, early treatment is essential. Preconception

treatment with LMWH, antioxidants and vitamins allows preventing
recurrent pregnancy complications and fetal losses in most cases.
Disclosure: Nothing to disclose.
C147
Atherogenic oxLDL/ b2GPI complexes and nitric oxide metabolites in
type 2 diabetes mellitus: baseline relationships and effect of rosuvastatin
treatment
Ames PRJ1, Garcia-De-La-Torre I2, Batuca J3, Kojima K4,
Matsuura E5, Lopez LR6
1
Airedale General Hospital, Steeton, Leeds, United Kingdom; 2Hospital
General de Occidente, Zapopan, Jalisco, Mexico; 3New University of
Lisbon, Lisbon, Portugal; 4Medical and Biological Laboratories Co.,
Ltd., Ina, Nagano, Japan; 5Okayama University Graduate School of
Medicine, Okayama, Okayama, Japan; 6Corgenix, Inc., Broomfield, CO
Oxidative stress and atherosclerosis characterise diabetes (DM):
atherogenic oxidated LDL/beta-2 glycoprotein I (oxLDL/ b2GPI)
complex is elevated in DM. In 76 type 2 DM patients (F 53, M 23,
age 5412 years, disease duration 7.77 years) we investigated the
relationship between baseline oxLDL/ b2GPI and nitric oxide metabo-
lites, nitrite (NO2), nitrate (NO3), nitrotyrosine (NT), asymmetric
dimethylarginine (ADMA) and the effect of 6 weeks oral treatment
with rosuvastatin 10 mg daily. At baseline oxLDL/ b2GPI significantly
associated with NO2 (r¼0.51, p<0.0001), NO3 (r¼0.42, p<0.0001),
ADMA (r¼0.27, p¼0.02) as well as with total cholesterol (r¼0.36,
p¼0.001), LDL (r¼0.39, p¼0.001), triglycerides (r¼0.4 p¼0.003) and
CRP (r¼0.26, p¼0.01). In addition, baseline NT positively related to
NO3 (r¼0.29, p¼0.01) and negatively to paraoxonase (r¼-0.38,
p¼0.0008). After rosuvastatin treatment oxLDL/ b2GPI decreased
by 44% (p<0.0001), ADMA by 5% (p¼0.07), NT and CRP were
essentially unchanged. Contrary to expectation NO2 decreased by
26% (p¼0.0003) and NO3 by 37% (p<0.0001). In type 2 DM athero-
genic oxLDL/ b2GPI relates to nitric oxide metabolites indirectly con-
firming its role as an atherogenic marker. In its surrogate role as a
marker of oxidant stress oxLDL/ b2GPI decrement after treatment
suggests that rosuvastatin may have antioxidant properties that may
be beneficial in DM. On the other hand, a decrease rather than an
increase of vasodilator nitric oxide (NO2) may be detrimental to vas-
cular function in DM.
Disclosure: Kazuo Kojima is employee of Medical and Biological
Laboratories Co., Ltd. Luis R Lopez is employee of Corgenix, Inc.
C148
Warfarin consumption and fibrin turnover are enhanced in thrombotic
patients with primary antiphospholipid syndrome
Ames P1, Ciampa A2, Ferrara F3, Iannaccone L3, Brancaccio V3
1
Airedale General Hospital, Steeton, West Yorkshire, United Kingdom;
2
Moscati Hospital, Avellino, Italy; 3Cardarelli Hospital, Napoli, Italy
Background: some patients with antiphospholipid syndrome require
unusually high doses of warfarin to attain their target international
normalized ratio (INR). Objectives: to investigate the possibility of
enhanced warfarin consumption in and fibrin turnover in thrombotic
primary antiphospholipid syndrome (PAPS) patients on oral
anticoagulation (OA) by longitudinally comparing their weekly
warfarin consumption with that of thrombotic patients with inherited
thrombophilia (IT) at similar target INR 2.0-3.0. Patients: Thrombotic
Abstracts
543
PAPS (n¼40, mean age 4416 years) and IT (n¼65, 5218) on OA
with warfarin followed up from June 1995 to June 2001. Methods: The
average weekly warfarin intake in milligrams (mg) was calculated by
dividing the total warfarin intake during the study period by the time
in weeks spent on oral anticoagulation. The average INR was calcu-
lated by summing all available INRs for individual patients and divid-
ing them by the number of visits over the study period. The
cytochrome CYP2C9 and the vitamin K epoxide reductase subunit 1
(VKORC1) were genotyped by PCR. IgG anticardiolipin antibody
(IgG aCL), IgG anti beta-2-glycoprotein-I (IgG b2GPI), protein
induced by vitamin K absence-factor II (prothrombin) (PIVKA-II)
and D-dimer (DD) were measured by immunoassay. Results: Age-
adjusted mean weekly warfarin consumption was greater in PAPS
than IT (27.62 vs 21.24 mg, p¼0.03); in PAPS baseline IgG aCL and
VKORC1 genotypes independently predicted weekly warfarin con-
sumption (p¼0.028 and p¼0.024); warfarin dose and IgG b2GPI inde-
pendently predicted PIVKA-II concentration (p¼0.02 and p¼0.01,
respectively). Age and sex adjusted DD was greater in PAPS than IT
(13234 vs 8337, p¼0.03). Conclusions: Warfarin consumption is
enhanced in thrombotic PAPS relative to IT and is partly accounted
by IgG aCL; despite oral anticoagulation PAPS show an enhanced
fibrin turnover that may have implications for re-thrombosis and
atherosclerosis.
Disclosure: Nothing to disclose.
C149
Risk factors for pregnancy failure in patients with antiphospholipid
syndrome treated with conventional therapies. A multicentre, case-
control study
Ruffatti A1, De Carolis S2, Rovere-Querini P3, Mosca M4, Mitic G5,
Tiziana Bertero M6, Pengo V1, Tincani A7
1
University of Padua, Padua, Italy; 2Catholic University of Sacred
Hearth, Rome, Italy; 3University of Milan, Milan, Italy; 4University of
Pisa, Pisa, Italy; 5University of Novi Sad, Novi Sad, Serbia and
Montenegro; 6Mauriziano Hospital, Turin, Italy; 7University of Brescia,
Brescia, Italy
Despite appropriate treatment, usually based on heparin and/or low
dose aspirin, about 20% of pregnancies in patients with antiphospho-
lipid syndrome (APS) are unsuccessful. A multicentre case-control
study was, thus, designed, to compare APS patients with successful
and unsuccessful pregnancy outcomes. The study group was made
up of 57 APS women who, despite conventional treatment, had unsuc-
cessful pregnancies and a control group of 57 APS women, matched
for age and treatment, whose pregnancies were successful. The clinical
and laboratory features of these patients were compared. The most
statistically relevant findings were: APS associated with autoimmune
diseases (OD¼6.9; CI 2.7 to 17.8), a history of both thrombosis and
pregnancy morbidity (OD¼12.7; CI 2.8 to 57.9), and lupus anticoagu-
lant plus anticardiolipin plus anti-beta2 Glycoprotein I antibodies
(triple) positivity (OD¼9.2; CI 2.8 to 30.9). The probability of preg-
nancy failure estimated by logistic regression analysis was 93% in APS
women with two or more of these features and 63% in those with only
one feature, while it dropped to 21% in women with none of these
features. It was found that APS patients, diagnosed on the basis of a
single positive test or on a history of pregnancy morbidity alone gen-
erally had successful pregnancies. It would seem from these findings
that the risk of failure in APS women planning a pregnancy can be
differentiated on the basis of their clinical and laboratory features.
Disclosure: Nothing to disclose.
Lupus
 Abstracts

544
C150 weeks since BHCGþ until week 34 of gestation. Results: Pregnancy
outcome of 528 women with APS:
Occasional antiphospholipid antibody positive patients also merit aspirin
therapy in recurrent pregnancy loss Aspirin þ
Sugiura-Ogasawara M1, Obayashi S1, Katano K1, Ozaki Y1 History without Aspirin þ LMWH þ
1
Nagoya City University, Graduate School of Medical Sciences, Table 1 treatment LWWH IVIG
Nagoya, Japan
N pregnancies 1546 671 25
It is well-known that treatment with aspirin plus heparin is effective for Live birth 290 (18.8%) 544 (81.1%) 25 (100%)
patients with antiphospholipid syndrome (APS) to prevent pregnancy Pregnancy loss 1256 (81.2%) 127 (18.9%) 0 (0%)
LPVC 213 (73.4%) 87 (135) 4 (16%)
loss. However, it is unclear if occasional antiphospholipid antibodies
(aPL) are a risk factor and whether patients with aPL at one time point
but not diagnosed as APS should be treated. We therefore studied Conclusion: In our experience, treatment with aspirin þ LMWH was
whether aspirin alone is effective in patients with occasional aPL associated with an improved gestational outcome. In selected patients,
who did not meet the criteria for APS. We compared live birth rates considered refractory to therapeutic standard, the addition of IVIG
between 52 patients with occasional aPL treated with aspirin and 672 would seem to improve the prognosis.
unexplained patients with no medication. Patients in both group had a
history of 2 or 3 pregnancy losses. Some 44 of 52 patients (84.6 %) with Disclosure: Nothing to disclose.
occasional aPL could experience live birth when treated with aspirin
alone. In contrast, 8 of 16 patients (50.0%) with occasional aPL could
have a baby without medication. Live birth rate in patients with occa- C152
sional aPL treated with aspirin was significantly higher than that in
The use of warfarin and low-dose aspirin in patients with
untreated patients with occasional aPL(p¼0.01). 509 of 672 patients
antiphospholipid syndrome
(75.7 %) with unexplained pregnancy losses could have babies. When
Kondratyeva L1, Reshetnyak T1
miscarriage cases caused by an abnormal embryonic karyotype were 1
State Institute of Rheumatology, Moscow, Russian Federation
excluded, the success rates were 95.7 % (44/46), 61.5 % (8/13) and
81.2 % (509/621), respectively. The live birth rate in patients with Objective: To compare thrombotic and hemorrhagic complications in
occasional aPL treated with aspirin was significantly higher than that patients with antiphospholipid syndrome (APS) before and during the
in unexplained patients with no medication (p¼0.008). moderate intensity warfarin therapy with or without aspirin. Material
We therefore conclude that aspirin is also useful in patients with and Methods: Of 71 patients with APS screened for 2,5 years in
occasional aPL but not APS. Institute of Rheumatology, a total of 60 patients were enrolled (45
Disclosure: Nothing to disclose. females, 15 males, mean age 37,811,6 years). Each patients’ history
was reviewed retrospectively. APS was associated with systemic lupus
erythematosus in 27 patients, and lupus-like disease in one patient, 32
C151 patients had primary form of syndrome. Warfarin therapy was started
to achieve an international normalized ratio of 2,0 to 3,0 (moderate
intensity) in all patients, 21 of them received the combination of war-
Treatment of the Antiphospholipid Syndrome During Pregnancy with
farin and low-dose aspirin. The efficacy of therapy was assessed by
Aspirin and Low Molecular Weight Heparin (LMWH) as Standard
recurrent thrombosis and transient ischemic attack (TIA), the safety
Treatment and Aspirin, Low Molecular Weight Heparin Plus
- by any bleeding. The rates of thrombotic or hemorrhagic events were
Intravenous Immunoglobulin (IVIG) in Refractory Cases
calculated as (total number of events 100) / total patients-years.
Sarto A1, Pappalardo C1, Pasqualini S1
1 Results: All patients had thromboses (arterial in 32 patients (53,3%),
Halitus Instituto Me´dico, Buenos Aires, Capital Federal, Argentina
venous in 41 patients (68,3%)) with the rate 19,6 cases per 100 patients-
Background: The Antiphospholipid Syndrome (APS) during preg- years and 16 (26,7%) patients had TIA with the rate 8 cases per 100
nancy is associated with recurrent pregnancy losses, fetal losses and patients-years before anticoagulant treatment. After warfarin starting
late placental vascular complications (LPVC). The clinical manage- our cohort was followed for a mean 16 months. Recurrent thrombosis
ment of pregnant woman with APS has advanced during the past occurred in 6 patients (7,6 cases per 100 patients-years), TIA – in 6
decade. Aspirin plus heparin reduce fetal loss rate in women with patients (7,6 cases per 100 patients-years) and major bleeding – in 4
APS, being the standard treatment nowadays. LMWH has replaced patients (5 cases per 100 patients-years). The cause for two episodes of
to unfractionated heparin (UFH) and can be administered safely to major bleeding was trauma.
pregnant women. Despite the use of these agents, many women still fail
to deliver a live. The association of aspirin, LMWH plus IVIG may be Rate of events
a therapeutic alternative in refractory cases. Objective: Describe our during anticoagulant Warfarin þ All
therapeutic experience in pregnant women with APS. Material and therapy (per 100 Warfarin low dose patients
patients-years) (n¼39) aspirin (n¼21) (n¼60)
Methods: We included 528 women who consulted in the Hematology
Department (1998-2008) and they had APS criteria by obstetric mor- Recurrent thrombosis 6 10,2 7,6
bidity history (Sapporo criteria). Standard treatment: 671 pregnancies Recurrent TIA 2 17,1 7,6
were treatment with aspirin and adjusted-doses enoxaparin (anti Xa: Major bleeding 0 13,7 5
0.3-0.8 U/ml). Treatment of refractory cases: The women considered
themselves refractory to the standard treatment who presented at least
one pregnancy loss with normal chromosomal embryo study and/or Conclusion: The moderate intensity warfarin therapy decreased the rate
fetal loss of more than 10 weeks and/or LPVC with maternal or peri- of recurrent thrombosis but no TIA in patients with APS. The combi-
natal risk of life. Twenty of those women had 25 pregnancies under nation of moderate – intensity warfarin therapy and low dose aspirin
treatment with aspirin, LMWH plus IVI 500 mg/kg one dose each 4 was not superior to moderate – intensity monotherapy of warfarin to

Lupus

prevent recurrent thrombosis in APS patients, but increased the risk of
serious bleeding.
Disclosure: Nothing to disclose.
C153
Intravenous immunoglobulin therapy of antiphospholipid syndrome in
relapsing cases despite standard treatment
Sciascia S1, Naretto C1, Rossi D1, Giachino O1, Alpa M1, Baldovino S1,
Roccatello D1
1
Centro Multidisciplinare di Immunopatologia e Documentazione su
Malattie Rare, CMID, Osp. G. Bosco, Torino, Italy
Background: Antiphospholipid syndrome (APS) is an autoimmune dis-
order defined by the occurrence of venous and arterial thromboses
and/or pregnancy morbidity, in the presence of antiphospholipid anti-
bodies. Data on the use of intravenous immunoglobulin (IVIg) in
patients with APS have focused on its obstetric complications, but
there are also case reports about treatments of other clinical manifes-
tations of the syndrome. Aim of the Study: To report our experiences in
the treatment with IVIg of relapsing despite standard therapy patients
with APS. Patients and Methods: Data from 44 APS patients were
investigated and in 7 subjects (16%) IVIg treatment was started
because of conventional therapy failure (low-weight-molecular heparin
and/or aspirin and/or warfarin and/or steroids). In 6 of 7 patients
(86%) recurrent thrombosis occurred and persistent thrombocytopenia
in 1 (14%) patient. IVIg (0,4 mg/kg/day) treatment was started (3
consecutive daily infusions for 3 months followed by one monthly
infusion for 9 months). In this study, main outcomes measures
included recurrence of thrombosis/thrombocytopenia, visual analog
scale (VAS) and health-related quality of life (HRQL) measured
before and after IVIg treatment. Testing for immunologic markers,
antiphospholipid antibodies profile (lupus anticoagulant, anticardioli-
pin and anti-b2GPI antibodies) were performed in all the patients.
Results: No further thrombosis occurred in patients with recurrent
episodes (mean follow up 70 months, range 6 - 108) and platelets
level were in range in the thrombocitemic patient (from 66.000 to
144.00/mm3). Only in 1 patients IVIg were suspended for the occur-
rence of sever macular-rash during the first IVIg cycle. In 6 of 7 patients
(86%) clinical improvement was observed after IVIg, as showed by
statistically significant difference between VAS values (p > 0.05) and
HRQL (p¼0,02) measured before and after IVIg treatment. No statis-
tically significant difference in antiphospholipid antibodies titer was
observed before and after IVIg treatment. Conclusion: Our data sug-
gest that IVIg may be an alternative therapy for relapsing cases of
antiphospholipid antibody syndrome.
Disclosure: Nothing to disclose.
C154
Procoagulant microparticles and vascular arterial disease in patients with
Systemic Lupus Erythematosus
Espinosa G, Patricio P, Tàssies D, Plası´n MA, Monteagudo J,
Cervera R, Reverter JC
Hospital Clinic, Barcelona, Spain
Background: Microparticles (MPs) are considered a key component in
the haemostatic response. Beyond their in vitro procoagulant proper-
ties a number of pieces of evidence points to procoagulant MPs as
efficient effectors in the haemostatic response, and as pathogenic mar-
kers of thrombotic disorders and vascular damage. Objectives: To
analyze the procoagulant activity of MPs and its correlation with the
Abstracts
545
development of arterial thrombosis and preclinical atherorsclerosis in
patients with systemic lupus erythematosus (SLE). Methods: We
included 100 patients with SLE (27 with associated APS, 36 with anti-
phospholipid antibodies (aPL) but without APS and 37 without aPL).
The presence of early atherosclerosis was evaluated by ultrasono-
graphic study of carotids measuring intima-media wall thickness and
presence of arteriosclerotic plaque. All thrombotic events were assessed
clinically and confirmed by objective methods. Procoagulant MPs were
assessed by a functional assay in which MPs were captured through
annexin V and then thrombin was formed by the addition of activated
factor X, activated factor V and prothrombin (Hyphen BioMed,
Neuville, France). Procoagulant MPs were expressed as nM phospha-
tidylserine equivalent. Results: A total of 16 episodes of arterial throm-
bosis in 8 patients with APS, 7 without aPL and 1 patient with aPL
have been registered. SLE patients with associated APS had greater
prevalence of plaque than patients without aPL or with aPL but with-
out APS (51.9% vs 24.3 % vs. 22.2 %). No differences were seen in
procoagulant MPs according the gender or the age of the patients.
Overall, there was a significant relationship between procoagulant
MPs and the presence of carotid plaque (16.7 8.6 nM vs 12.7 7.3
nM; p¼0.02) and the number of carotid plaques (14.5 4.0 nM in
patients with one plaque and 17.3 10.3 nM in patients with 2 or
more plaques; p¼0.02). In addition, a relationship between thrombotic
events and the procoagulant activity (18.2 9.2 nM with events vs 12.7
6.9 nM: p¼0.002 without events). Interestingly, this higher amount
of procoagulant activity in patients with thrombosis was due to higher
procoagulant activity in patients with arterial thrombosis versus those
without (18.7 9.5 nM vs 13.0 7.2 nM; p¼0.007). Conclusions:
Procoagulant MPs may be implicated in preclinical arteriosclerosis
and arterial thrombosis in SLE patients.
C155
Antiphospholipid syndrome-associated nephropathy in patients with
systemic lupus erythematosus: prevalence, clinical manifestations and
renal functional outcome
Bucciarelli S, Silvariño R, Pons-Estel G, Espinosa G, Arrizabalaga P,
Sant F, Sole M, Cervera R
Hospital Clinic, Barcelona, Catalonia, Spain
Background: The presence of antiphospholipid antibodies (aPL) has
been associated with small vessel renal injury and chronic renal ische-
mia. The renal pathological findings associated with antiphospholipid
syndrome (APS) include thrombotic microangiopathy (TMA), repre-
senting an acute lesion, and chronic renal damage, represented by
fibrous intimal hyperplasia (FIH), fibrocellular and fibrous arterial
and arteriolar occlusions (FAO), focal cortical atrophy (FCA) and
tubular thyroidization (TT). The common clinical manifestations are
hypertension, acute renal failure or chronic low-grade proteinuria.
Although the term ‘‘APS-associated nephropathy’’ has been suggested,
renal involvement has not yet included within the classification criteria
of APS. Objectives: To determine the prevalence, clinical manifesta-
tions and renal functional outcome of patients with lupus nephritis and
APS-associated nephropathy. Methods: Patients were included if the
following criteria were present: 1) a diagnosis of SLE (ACR criteria)
nephritis confirmed by renal biopsy; 2) patients with systemic sclerosis,
hemolytic-uremic syndrome, systemic vasculitis, thrombotic thrombo-
cytopenic purpura, diabetic nephropathy, preeclampsia, and infection
with human immunodeficiency virus were excluded. The histological
samples were all analyzed by the same pathologist. Results: Seventy-
nine biopsies were included (70 female [88.6%]). The mean age at time
of biopsy was 33.3  16.6 years (range, 14-66). The follow-up period
(time between the renal biopsy and the last medical visit) was 73  51
months (range, 1-215). Nine (11.4%) renal biopsies met diagnostic
criteria for APS nephropathy. Three (33.3%) showed acute APS
Lupus
 Abstracts
546
nephropathy. The following histological lesions were found: 3 (3.8%)
cases of TMA, 4 (5%) of FIH, and 3 (3.8%) cases of FCA. The SLE
patients with APS nephropathy had a higher duration of SLE at the
time of renal biopsy (p¼0.02) and higher rate of chronicity (p¼0.01).
They also had higher prevalence of interstitial fibrosis (p¼0.04),
tubular atrophy (p¼0.02) and nephro-angiosclerosis (p¼0.03) on his-
topathology when compared with the group without APS nephropa-
thy. The presence of aPL was associated to APS nephropathy
(p¼0.009). Association of positive lupus anticoagulant and anticardio-
lipin (but not isolated positivity) was related to the development of
APS nephropathy (p<0.001). There was no association between APS
nephropathy and the presence of hypertension, nephrotic syndrome,
hematuria, proteinuria or elevated serum creatinine levels. No signifi-
cant difference in complete renal response, partial renal response and
no response between groups were found. Patients with TMA showed
higher prevalence of APS (p<0.001), extrarenal arterial thrombosis
(p¼0.02) and venous thrombosis (p¼0.02) during their follow-up.
Conclusion: The prevalence of APS-associated nephropathy was
11.4%. There was no relationship between APS-associated nephropa-
thy and any clinical manifestation or laboratory feature except for
increased risk to develop APS and arterial and venous thrombosis.
The long-term renal outcome was similar in SLE patients with or with-
out APS-associated nephropathy.
Disclosure of Interest: None declared.
C156
Antiphospholipid Antibodies and Recurrent Pregnancy Loss: Prevalence
of Anticardiolipin antibodies, the lupus anticoagulant, and
antiphosphatidyl serine antibodies
Kutteh WH, Corey A, Jaslow CR
University of Tennessee Health Sciences Center, Memphis, Department
of Biology, Rhodes College, Memphis, TN. USA
Objective: To describe the prevalence of anticardiolipin antibodies
(ACL), the lupus anticoagulant (LAC), and antiphosphatidyl serine
antibodies (APS) in women who had suffered two, three, or four or
more consecutive recurrent pregnancy losses (RPL). Patients: Included
872 women with RPL. The majority of women (96%) had losses that
occurred before 13 weeks. The EGA at loss was 8.6  3.6 weeks. All
women had a complete evaluation including karyotypes, uterine anat-
omy, LAC, ACL, APS, TSH, prolactin, and Factor V Leiden. Control
women were 288 nonpregnant, reproductive-aged, parous women
without a history of adverse pregnancy outcome. Methods: Serum
levels of ACL and APS IgG and IgM were measured by enzyme-
linked immunosorbant assay, with referenced standards and known
positive and negative sera on each plate. Abnormal levels exceeding
20 phospholipid units for ACL or 3.0 multiples of the median for APS
(the 99th percentile of a normal population) were confirmed by repeat
testing at least 6 weeks later. Serum levels of LAC were evaluated
using the Russell viper venom test and PTT-LA. Results >42 seconds
that were not corrected with a 1:1 mix with normal serum were con-
sidered positive if confirmed by a hexagonal phase phospholipid test.
Results: In women with RPL, positive ACL were detected in 132/872
(15.1%), LAC in 31/872 (3.6%), and APS in49/872 (5.6%). APS
were identified in women with RPL who tested negative for ACL.
APS were identified in women with two losses {18/391 (4.6%)},
Lupus
three losses {16/288 (5.6%)}, and in women with four or more losses
{15/193 (7.8%)}. In control women, positive ACL were detected in 14/
288 (4.9%), LAC in 3/288 (1.0%), and APS in 8/288 (2.8%).
Differences in ACL and APS were significant using the two-tailed
Fisher’s exact test. Conclusions: APS are known to inhibit trophoblast
development and invasion, retard syncytiotrophoblast formation, and
decrease the synthesis of hCG. In vitro, heparins have been shown to
reduce the binding of APS to substrate. Several clinical trials of women
with RPL and positive APS have suggested encouraging results in
successful outcome after treatment with heparin. Diagnostic tests for
APS may be useful when evaluating patients with RPL.
C157
Significance of Anticardiolipin antibodies in patients with lupus nephritis
Umer S, Hayat S, Berney SM
Louisiana State University, Shriveport, LA, USA
Background: Systemic Lupus Erythematosus is characterized by the
presence of auto antibodies such as antiphospholipid antibodies. We
observed elevated antiphospholipid antibodies in a significant number
of our lupus nephritis (LN) patients. As a result, we sought to quantify
these antiphospholipid antibodies in our lupus nephritis patients and
correlate them with the levels of anti-double stranded DNA antibodies
(anti- DS DNA ab) and C3 and C4. Methods: We reviewed the med-
ical records of Systemic Lupus Erythematosus patients with kidney
biopsies followed at Louisiana State University Health Sciences
Center, Shreveport from 1996–2007 for this retrospective study and
analyzed the levels of antiphospholipid antibodies with anti DS-
DNA ab , C3, C4 and nephritis activity. Results: Sixty five patients
(11 males + 54 females; 61 African American + 4 Caucasians) were
included in this study. 25 patients had World Health Organization type
4 Lupus Nephritis, 28 had type 3 Lupus Nephritis and 12 had Type 5
Lupus Nephritis. The patients ages range was from 5 to 66 years.
Among those with type 4 LN, 16 of the 25 (64%) were positive for
anticardiolipin antibodies at the time of renal biopsy. 18 of the 28
(64%) patients with type 3 lupus nephritis were positive for anticar-
diolipin antibodies at time of renal biopsy and 6 of the 12 (50%)
patients with type 5 Lupus Nephritis were positive for ACL antibodies
at the time of renal biopsy. 10 of 65 patients were not checked for
anticardiolipin antibodies at the time of renal biopsy. Low C3 and
C4 levels were found in 55 of the 65 patients. Anti DS DNA ab were
elevated in 51 patients with titers ranging from 30 to 2932 (normal ¼
<25 IU). All patients with type 3 and 4 were treated with intravenous
Cyclophosphamide (6–10 doses) followed by either Mycophenolate
Mofetil or Azathioprine. 31 of 65 patients achieved complete remis-
sion. Among 40 patients with positive anticardiolipin antibodies, 13
progressed to ESRD despite cytotoxic therapy, 8 developed CKD after
treatment and 19 achieved remission. Conclusions: This retrospective
study suggests that elevated level of anticardiolipin antibodies appears
to be associated with Lupus Nephritis and may correlate with comple-
ment consumption indicating increased disease activity.
Anticardiolipin antibodies may represent a complementary marker
for lupus activity in addition to anti Double Stranded DNA ab in
our patients with lupus nephritis and may also indicate patients who
may have a worse renal outcome. Further prospective studies are
required to investigate this association.