MUST TO KNOW IN IMMUNOHEMATOLOGY (BLOOD BANKING)

ISBT 001 ABO

ISBT 002 MNS
ISBT 003 P
ISBT 004 Rh
ISBT 005 Lutheran
ISBT 006 Kell
ISBT 007 Lewis
ISBT 008 Duffy
ISBT 009 Kidd
ISBT 010 Diego
ISBT 011 Cartwright
ISBT 012 Xg
ISBT 013 Scianna
ISBT 014 Dombrock
ISBT 015 Colton
ISBT 016 Landsteiner-Weiner
ISBT 017 Chido/Rodgers
ISBT 018 H
ISBT 019 Kx
ISBT 020 Gerbich
ISBT 021 Cromer
ISBT 022 Knops
ISBT 023 Indian
Chromosome 1 Rh
Duffy
Scianna
Cromer
Knops
Chromosome 2 Gerbich
Chromosome 4 MNS
Chromosome 6 Chido/Rodgers
Chromosome 7 Cartwright
Colton
Kell
Chromosome 9 ABO
Chromosome 11 Indian
Chromosome 17 Diego
Chromosome 18 Kidd
Chromosome 19 H
Lewis
Landsteiner-Weiner
Lutheran
Chromosome 22 P
Chromosome X Xg
Kx
Chromosome: Not known Dombrock
Von Descatello AB
(Decastello)
Sturle (Sturli)
Blood groups (Most common) O > A > B > AB (Least common)

Cell typing Forward/direct typing

Specimen: RBCs (Ag)
Reagents:
= Anti-A (blue: trypan blue)
= Anti-B (yellow: acroflavin dye)
= Anti-AB (colorless)
Uses of anti-AB Checks for the reaction of anti-A and anti-B
Detects weak subgroup of A and B because it has higher titer of anti-A & anti-B
Serum typing Reverse/indirect typing
Specimen: Serum (Ab) = 3-6 months (development)
Reagents:
 = A1 cells
= B cells
Gel typing Major Advantage: Standardization
Medium: Dextran-acrylamide gel
0 = Unagglutinated Cells ---(media)---> Bottom of the µtube (microtube)
1+ = Agglutinated cells ---(media)---> May concentrate at the bottom of µtube
2+ = Agglutinated cells ---(media)---> Throughout the length of the µtube
3+ = Agglutinated cells ---(media)---> Concentrate near the top of the µtube
4+ = Agglutinated cells ---(media)---> Top of the media
Mixed-field:
= Agglutinated cells ---(media)---> Top
= Unagglutinated cells ---(media)---> Bottom
Red cell Ag-Ab reactions 0 = No agglutination/hemolysis
W+ = Tiny agglutinates, turbid BG
1+ = Small agglutinates, turbid BG (25% agglutination)
2+ = Medium agglutinates, clear BG (50% agglutination)
3+ = Large agglutinate, clear BG (75% agglutination)
4+ = One solid agglutinate (100% agglutination)
Universal donor Group O
Universal recipient Group AB
Universal donor for RBCs Group O (No Ag)
Universal recipient for Group AB (No Ab)
RBCs
Universal donor for Group AB (No Ab)
plasma
Universal recipient for Group AB (No Ag)
plasma
RCS: Red cell suspension BB = 2-5% RCS
Approximate = Tomato red
Exact % RCS = [Vol. RBCs (mL) ÷ TV (mL)] x 100
Genotype One’s genetic makeup
Phenotype Expression of one’s genes
Homozygous In double dose
Heterozygous In single dose
Dominant Always expressed even in the heterozygous state
Recessive Not expressed when (+) dominant gene
To be expressed, it should be in the homozygous state
Allele One of two or more different genes that may occupy a specific locus on a chromosome
Silent/amorph Only indicates the absence of the Ag
Von Dungern Subgroups of A = A1 (80%) and A2 (20%)
Hirzfeld
Dolichos biflorus Lectin source of anti-A1
Phenotype Possible Genotypes
A1 A1A1
A1A2
A1O
A2 A2A2
A2O
A 1B A1B
A 2B A2B
B BB
BO
O OO
Gene Glycosyltransferase Immunodominant Ag Acceptor molecule
Sugar
H L-fucosyltransferase L-fucose H Precursor subs.
A N-acetylgalactosaminyltransferase N-acetyl-D- A H
galactosamine
B D-galactosyltransferase D-galactose B H
AB N-acetylgalactosaminyltransferase N-acetyl-D- A H
D-galactosyltransferase galactosamine B
D-galactose
 O -- -- -- Unchanged H
Amount of H Ag (Greatest) O > A2 > B > A2B > A1 > A1B (Least)
Bombay individual Bhende
(-) H gene
hh or Hnull
Lack H, A and B Ag’s
Designated as Oh
w/ anti-H, anti-A and anti-B Ab’s
Mistyped as group O
Parabombay Inherit weak H gene
w/ detectable A and B Ag’s but no detectable H Ag
Ah, Bh, ABh
Anti-H Differentiates Group O from Oh individuals
Ulex europaeus Lectin source of anti-H
ABO Histoblood group = present on all tissues and organs of the body
May be expressed in secretions depending on the secretor status (SeSe or Sese)
Determination of Secretor Specimen: Saliva
Status Principle: Hemagglutination-inhibition
ABO antibodies Mixture of IgM, IgG and IgA (Henry)
Anti-A Predominantly IgM
Anti-B React at room temp
Naturally occurring
Anti-A,B (Group O) Predominantly IgG
React at 37’C
Immune Ab
Immune Ab’s Production is stimulated by:
1. Pregnancy
2. Incompatible transfusion and transplant
ABO HDN Mother: Group O
Child: Group A or B
Group I discrepancies Weak reacting/missing Ab’s
Newborns
Elderly patients
Hypogammaglobulinemia/agammaglobulinemia
Group II discrepancies Weak reacting/missing Ag’s (Less frequent)
Leukemia
Acquired B phenomenon
Subgroups of A
Hodgkin’s disease
BGSS (Remedy: Wash RBCs)
Group III discrepancies Plasma abnormalities resulting to Rouleaux formation (Plasma factor)
á globulins: MM, WM, HL
á fibrinogen
Dextran and PVP
Wharton’s jelly (Remedy: Wash cord cells 6-8x (7x)
Group IV Miscellaneous:
Polyagglutination
Anti-I (cold agglutinin)
Cis “AB phenotype”
Unexpected ABO isoagglutinins:
= Anti-H: produced by A1 and A1B (âH Ag)
= Anti-A1: produced by A2 and A2B (No A1 Ag)
 A Subgroups
A3 MF agglutination w/ anti-A & anti-A,B
Ax Weak MF agglutination w/ anti-A& anti-A,B
Aend MF agglutination w/ anti-A,B
Am (-) Agglutination, (+) AH substance in secretions
Ay (-) Agglutination, â AH substance in secretions
Ael (-) Agglutination, (+) H substance in secretions
B Subgroups
B3 MF agglutination w/ anti-B & anti-A,B
Bx MF agglutination w/ anti-A,B
Bm (-) Agglutination, (+) BH substance in secretions
Bel (-) Agglutination, (+) H substance in secretions
Rh Nomenclatures 1. Rosenfield = no genetic basis, indicates the presence or absence of Rh Ag’s
2. Fisher-Race (DCE) = inheritance of 3 genes (d= amorph/silent)
3. Wiener (Rh-hr) = inheritance of 1 gene (Ex. R0), w/c codes for an agglutinogen (Ex.
Rh0), w/c contains 3 blood factors (Ex. Rh0hr’hr’’)
Immunogenicity of Rh (Most immunogenic) D > c > E > C > e (Least immunogenic)

Ag’s
R or r D or d
1 or ‘ Ce
2 or ‘’ cE
Z or y CE
G D+C+ red cell
Rh: 13, 14, 15, 16 4 different parts of the D mosaic
Genetic weak D D Ag’s expressed appear to be complete, but few in number
D Mosaic (Partial D) If one or more parts of D Ag is missing à weakened expression of D Ag
May produce anti-D (Ab against missing fragment)
4 fragments
C Trans D ---(trans)---> C (Ex. Dce/dCe)
f (ce) c ---(cis)---> e [Ex. Dce/DCE]
rhi (Ce) C ---(cis)---> e [Ex. DCe/DcE]
Hr0 “Common” Rh phenotypes (R1R1, R2R2, rr)
Rh Ab’s IgG1 and IgG3
React at 37’C
Immune Ab’s
(-) C’ binding = extravascular hemolysis (delayed HTR)
Rh HDN Mother: Rh (-)
Child: Rh (+), 2nd pregnancy
RhoGam or RhIg Purified anti-D
Administer w/in 72 hrs after 1st delivery
Full dose RhoGam 300 µg anti-D
(>12 weeks gestation) Protect up to 30mL D+ WB or 15mL D+ RBCs
Minidose/Microdose 50 µg anti-D
RhoGam Protect up to 5mL of D+ WB or 2.5mL D+ RBCs
(<12 weeks gestation) Ex. Abortion
# RhIg vials Volume of FMH (mL) ÷ 30
Vol. FMH = % fetal cells x 50
---------------------------------------------
x = (% Fetal cells x 50) ÷ 30
x ≈ __ + 1 = # RhIg vials
As little as 1mL Rh(+) Produces anti-D
RBC
Rh+ RBCs + anti-D = (+) agglutination
Perform test for Du (IAT) If RBCs + anti-D = (-) agglutination
= IAT is (+) agglutination = +Du (weak D)
Rh- RBCs + anti-D + AHG reagent = (-) agglutination
2 conditions wherein an 1. No prior exposure to D Ag (males) or past childbearing age (females)
Rh- pt. can be transfused 2. Administer RhoGam
w/ Rh+ blood
Anti-LW Originally identified as anti-Rh in early experiments involving rabbits immunized w/
(Landsteiner-Weiner) Rhesus monkey blood
Anti-LW agglutinates Rh+ and Rh- cells except Rhnull cells
Rhnull No Rh Ag
Rhnull
Designated as ---/---
Stomatocytes
Rhdeleted No C/c and E/e Ag
Designated as D--/D--
Lewis system Le gene codes for the production of fucosyltransferase enzyme that catalyzes addition of
fucose to the 4th C of N-acetylglucosamine of type 1 precursor
Lewis Ag’s Lea ---(Se)---> Leb
Produced by tissue cells
Not well developed at birth = NB/cord cells = Le(a-b-)
Decreased expression during pregnancy
Genotype Substances (Secretion) Phenotype Le Ab’s
ABH, lele, sese None ABH, Le(a-b-) Anti-Lea & Anti-Leb
ABH, lele, SeSe/Sese ABH ABH, Le(a-b-) Anti-Lea & Anti-Leb
ABH, LeLe/Lele, sese Le a ABH, Le(a+b-) Anti-Leb
ABH, LeLe/Lele, ABH, Lea, Leb ABH, Le(a-b+) none
SeSe/Sese
Lewis Ab’s Anti-Lea & Anti-Leb
Naturally occurring
IgM
Activates the C’
MN Ag’s Glycophorin A (MN-SGP)
M = Ser-Ser-Threo-Threo-Gly
N = Leu-Ser-Threo-Threo-Glu
Well developed at birth
Important in paternity testing
Anti-M IgM, pH-dependent (6.5), glucose-dependent
Anti-N-like Ab IgM
Found in renal patients dialyzed w/ formaldehyde sterilized equipment
SS Ag’s Glycophorin B (Ss-SGP)
S = Methionine (29th)
s = Threonine (29th)
Ss Ab’s IgG
React at 37’C and AHG
Severe HTR w/ hemoglobinuria and HDN
Phenotype Detectable Ag’s Possible Ab’s
P1 P1 and P None
P2 P Anti-P1
p (p null) None Anti-P, P1, Pk (anti-Tja)
P1k Pk and P1 Anti-P
P2k
Pk Anti-P, anti-P1
P1-like Plasma, pigeon and turtledove droppings, turtledove eggwhite
P1 substance Hydatid cyst fluid, Lumbricoides terrestris, Ascaris suum
Anti-P1 IgM
Naturally occurring
Strong anti-P1 = Hydatid disease (E. granulosus)
Associated w/ fascioliasis, C. sinensis and O. viverrini infections
Anti-Tja Anti-P, P1, Pk
Spontaneous abortions in early pregnancy
Anti-P IgG
Naturally occurring
Biphasic hemolysin (PCH)
Test: Donath-Landsteiner
Ii Blood Group I: Individuality
Neonates = âI áI (Ag) = I-i+
Adults (18 mos.-adult) = áI âI = I+i-
HEMPAS ái Ag in adults
Anti-I Interfere w/ reverse typing (Group IV)
Benign anti-I = normal, IgM, naturally occurring, react at 4’C
Pathologic anti-I = IgM, react at 30/32’C (CAS = PAP)
Autoanti-I = M. pneumoniae, L. monocytogenes
Anti-i IgM
React at 4’C
EBV caused
Diseases of RES:
 - Alcoholic cirrhosis
- Myeloid leukemia
- Reticuloses
Anti-IT Transition: from i à I
Yanomama Indians
Hodgkin’s lymphoma
K Kell
k Cellano
Kpa Penney
Kpb Rautenberg
Js a Sutter
Js b Matthews
Kell Ag’s Immunogenicity: 2nd to D (D>K>c>E>C>e)
Synthesized on precursor Kx
= On WBCs: remain unconverted. If (-) à CGD
= On RBCs: converted to Kell Ag’s. If (-) à MacLeod phenotype
MacLeod phenotype Acanthocytosis
Muscular dystrophy

Anti-K IgG
React at 37’C and AHG
Fy(a+b-) Chinese (90.8%)

Fy(a-b-) American blacks

Resistant to P. vivax/P. knowlesi (malaria)
Fya and Fyb Receptors for malaria
Duffy gene 1st human gene to be assigned to specific chromosome (1)
Anti-Fya IgG
Anti-Fyb React at AHG
Jka and Jkb Enhanced by enzymes
Anti-Jka Notorious antibody = not easily detected
Anti-Jkb Immune Ab’s (IgG)
Common cause of delayed HTR
Autoanti-Jk = Methyldopa (Aldomet)
Lua and Lub Development at age of 15
Anti-Lua IgG, IgM, IgA
Naturally occurring
React at room temp
Anti-Lub IgG4, IgM, IgA
Immune Ab
React at 37’C
Diego (Di) Mongolian ancestry
AE-1 = HCO3- <--> Cl-
Defect in AE molecule “ASO”
Acanthocytosis
Hereditary Spherocytosis
SEA Ovalocytosis
Cartwright (Yt) Erythrocyte acetylcholinesterase = neurotransmission
Xg Sex-linked
áá Females
Short arm of X chromosome
Scianna Sc1, Sc2, Sc3
Dombrock Gregory (Gya)
Holley (Hy)
Joseph (Joa)
Colton (Co) CHIP, Aquaporin = water permeability
Chido/Rodgers (Ch/Rg) HLA system (Bg)
C4A (Rg) and C4B (Ch) = C’ component
HTLA = exhibit reaction only at high dilution
DAF (Cr)
Gerbich (Ge) GPC and GPD
Leach phenotype (GE: -2, -3, -4) = Elliptocytosis
Cromer (Cr) DAF
CROM Ab’s = black individuals
Knops (Kn) CR1 (C’ receptor 1)
Indian (In) CD44 (immune adhesion)
Benneth-Goodspeed (Bg) HLA Ag’s on RBCs (Class I MHC)
Bga = HLA-A7
Bgb = HLA-A17
Bgc = HLA-B28
Public Ag High-incidence Ag’s
Ena (99.9%)
Private Ag Low-incidence Ag’s (<1%)
HTLA Ab’s Anti-Ch
Anti-Rg
Anti-Kn
Anti-Yka
Anti-Csa
Anti-McC
Anti-JMH
Sources of Substances for Neutralization for Certain Antibodies
Anti-P1 Hydatid cyst fluid, pigeon droppings, turtledoves’ egg whites
Anti-Le Plasma/serum, secretor saliva
Anti-Ch, Anti-Rg Plasma/serum
Anti-Sd a Urine, guinea pig urine
Anti-I Mother’s milk
Anti-H Saliva
Effect of Proteolytic Enzymes on Select Antigen-Antibody Reactions
Enhanced P1
Rh
I
Kidd
Lewis
Destroyed MNS
Duffy
Chido
Rodgers
Cartwright
Xg
Donation Process
At least 2 persons Bleeder
Head (BB)
Donor registration 1st step
Interview & physical 2nd step
exam
Actual donor selection and 3rd step
blood collection
Collection <15 mins (7-10mins)
>15 mins = (-) cryoprecipitate
Donor bleeding 450 angle à 10-200 angle
(Venipuncture: 150 angle)
Basic Qualifications of the Potential Blood Donor
Good health Purplish blue lesions à Kaposi’s sarcoma (HIV, HHV-8)
Age 18-65 yrs old
<18 yrs old = w/ parent’s consent
>65 yrs old = w/ physician’s consent
Body weight Max: 10.5mL/kg
Ideal: 110 lbs (50kg)
450mL blood + 30mL blood (serologic tests)
63mL anticoagulant

Donor <110 lbs (<50kg):

=â Volume of blood to be drawn
=â Volume of anticoagulant
Vol. of blood to be drawn A = (Donor weight ÷ Ideal weight) x 450mL
Vol. of anticoag. needed B = (A ÷ 100) x 14
Vol. of anticoag. to be C = 63 – B
removed from blood bag
1:7 Anticoagulant: Blood ratio (Blood bag)
63mL AC = 450mL blood
31.5mL AC = 200mL blood
Temperature 37.5’C (99.5’F)
Pulse 50-100 beats/min (â athletes = acceptable)
BP cuff 40-60 mmHg (if used as tourniquet)
BP (DOH) 90-160 mmHg
60-100 mmHg
Min. H & H Allogeneic donation:
Hgb: ≥12.5 g/dL
Hct: ≥38%
Autologous donation:
Hgb: ≥11 g/dL
Hct: ≥33%
CuSO4 method Hgb determination
30mL container = 25 tests, solution changed daily
SG: CuSO4 = 1.053
1 cm = distance between blood and solution
Acceptable drop of blood à sink w/in 15 secs (Hgb: ≥12.5 g/dL)
Donor Deferral
Permanent Chagas’ disease

Babesiosis
Tegison (Tx: Psoriasis) = teratogenic
Recipient of human (pituitary)-derived GH = risk of transmitting CJD
[Recombinant GH = no deferral]
Recipient of cornea/dura mater = risk of transmitting CJD
3 years Malaria refugee/immigrant
1 year (12 months) Recipients of blood known to be possible sources of hepatitis
Tattoo
Rape
Incarceration in jail (3 days/72 hrs)
Blood transfusion
Major operation including dental surgery
Syphilis
Gonorrhea
Traveler à malaria-endemic places
Rabies vaccine
9 months (DOH) Childbirth (AABB: 6 weeks)
3 months (12 weeks) Recent blood donation (AABB: 8 weeks)
[DOH] = 450mL blood: 12 weeks
= 200mL blood: 6-8 weeks
1 month (4 weeks) Rubella vaccine
Isotretinoin/Accutane (Tx: Acne) = teratogenic
Proscar (Tx: Benign prostatic hyperplasia) = teratogenic
2-3 weeks After febrile episodes
2 weeks Rubeola vaccine
Polio vaccine
Mumps vaccine
2 days (48 hrs) After hemapheresis
12 hrs After alcohol intake
Cause: Arterial puncture Jet-like pulsating bleeding w/ bright red blood
Citrate Binds Ca2+
Massive transfusion (8-10 units) à Citrate toxicity à Hypocalcemia
Milk = áCa2+
Dextrose/Glucose Provides energy for red cells
Citric acid Prevents caramelization (âpH)
Adenine For improved survival of red cells
Phosphate buffer á ATP
Shelf-life: 21 days ACD (Apheresis)
CPD
CP2D
Shelf-life: 35 days CPDA-1
Shelf-life: 42 days CPDA-2 (DOH)
Additive solutions:
-Adsol (AS-1)
-Nutricel (AS-3)
 -Optisol (AS-5)
Additive solutions SAGM:
-Saline
-Adenine
-Glucose
-Mannitol = RBC membrane stabilizing agent
Rejuvenation solutions á ATP & 2,3-DPG (Rejuvesol)
PIGPA = Phosphate, Inosine, Glucose, Pyruvate, Adenine
PIPA = Phosphate, Inosine, Pyruvate, Adenine
Heavy spin 5000g for 5mins = pRBC, platelet concentrate
5000g for 7mins = cryo, cell-free plasma
Light spin 2000g for 3mins = platelet-rich plasma
6-8 hours processing To maximize the number of components derived from 1 unit of blood
Platelet concentrate Light spin à Heavy spin
Centrifuge at 20-24’C
[Other blood components: Ref. centrifuge (1-6’C)]
áBacterial contamination
Separate from WB w/in 6-8 hrs
Blood Component Indication Storage Transport Shelf-life Other Info
Whole blood 1. Blood vol. expansion & 1-6’C 1-10’C 21d (ACD, CPD) 1 unit:
á RBC mass 35d (CPDA-1) áHgb by 1 to
42d (CPDA-2)
2. Massive transfusion & 2d (Heparin 1.5g/dL
acute blood loss áHct by 3-5%
Packed RBCs 1. á RBC mass 1-6’C 1-10’C Open system: 24h ‡
80% plasma
(Normovolemic patients) Closed system: removed
21d (ACD, CPD) ‡
(Hct: 65-80% but
35d (CPDA-1) not >80%)
1 unit:
áHgb by 1 to
1.5g/dL
áHct by 3-5%
Blood Component Indication Storage Transport Shelf-life Other Info
Leukoreduced RBCs 1. FNHTR (Leuko. Ab’s) = 1-6’C 1-10’C Same as WB Preparation:
á1’C in temp. 1. Centrifugation
2. Prevent CMV 2. Filtration
3. Saline washing
Washed RBCs 1. Allergic (plasma w/ 1-6’C 24h Also for
foreign protein) polyagglutination
2. FNHTR (Leuko. Ab’s)
Anaphylactic: âIgA w/
anti-Iga
Frozen RBCs 1. Prolong storage of rare -65’C or 10 years
units -125’C
2. Prolong storage of
autologous units
Deglycerolized RBCs 24h
Fresh frozen plasma 1. Multiple coag. factor def. -18’C 1 year (-18’C) ‡
Contains plasma,
(liver disease) -65’C 7 years (-65’C) all coag. factors and
2. Replace isolated factor complement
def. when specific ‡
FFP: Thaw at 37’C
component is not available ‡
Thawed plasma:
3. Reverse effects of
store at 1-6’C
coumarin/warfarin ‡
Administer w/in
24hrs once thawed
Cryoprecipitate 1. Fibrinogen def. -18’C or 1 year ‡
Contains:
2. Hemophilia A colder -150mg fibrinogen
3. vWD -80 U AHF
4. Factor XIII def. -vWF
-Factor XIII
‡
Cryoppt.: Thaw at
37’C
‡
Thawed cryoppt
(<15mL; not group-
spec.): store at RT’
‡
Administer w/in 6
hrs
Granulocyte 1. Granulocyte dysfunction 20-24’C 24h Contains 1 x 1010
concentrate (CGD) w/o granulocytes
2. Myeloid hypoplasia agitation
Platelet concentrate 1. Thrombocytopenia 20-24’C 5d (20-24’C w/ 1 unit will á platelet
w/ agitation) count by 5,000-
agitation 2d (1-6’C) 10,000/µL
1-6’C
Irradiated blood 1. Prevent GVHD 28 days from Usually RBCs and
component irradiation or orig. platelets are
exp. date whichever irradiated
comes first
Factor VIII 1. For hemophilia A 1-6’C Varies Stored in lyophilized
concentrate form
Factor IX concentrate 1. For hemophilia B 1-6’C Varies ‡
A.k.a. prothrombin
complex
‡
Contains factors II,
VII, IX and X
NSA 1. Hypovolemic shock 2-10’C 5 years 96% Albumin
4% Globulin
Blood Component Indication Storage Transport Shelf-life Other Info
PPF 1. Hypovolemic shock 2-10’C 5 years 80-85% Albumin
10-15% Globulin
SDP 5 years SDP: Single donor
plasma
Methods of Freezing RBCs
Concentration Frozen at Stored at Equipment
High glycerol 40% w/v glycerol -80’C -65’C Mechanical
(Slow freezing) freezer
Low glycerol 20% w/v glycerol -196’C -120’C Liquid nitrogen
(Fast freezing)
Agglomeration Glycerol -80’C -65’C Mechanical
Glucose freezer
Fructose
EDTA
Cryoprotective agent Prevents rupture of RBCs during freezing
Ex. Glycerol
Deglycerolization Removal of glycerol
Hypertonic solution followed by an isotonic solution
High glycerol (DG) 12% NaCl -----> 1.6% NaCl -----> 0.9% NaCl
Low glycerol (DG) 45% NaCl in 15% mannitol
Agglomeration (DG) 50% Glucose + 5% Fructose -----> 0.9% NaCl
Cryoprecipitate After thawing = administer w/in 6 hours
After pooling = administer w/in 4 hours
Leukapheresis HES (Hydroxyethylstarch) = áSeparation bet. WBCs and RBCs
Donor: administered w/ corticosteroids 12-24 hrs before donation
= á# of circulating granulocytes
Plateletpheresis Usually takes 1-2 hours
Donor:
= Platelet count: ≥150 x 109/L
= Aspirin-free: 3 days
Single donor platelets A.k.a. super packed platelets
(SDP) From plateletpheresis
For patients who are refractory or unresponsive to RDP
Limit patient exposure to multiple donors
RDP: Random Donor Platelets SDP: Single Donor Platelets
Preparation From WB: Light spin à Heavy spin Plateletpheresis
Amount 5.5 x 1010 platelets 3.0 x 1011 platelets
Vol. of plasma 50-75mL 300mL
pH ≥ 6.0 (New: 6.2) ≥6.0 (New: 6.2)
Storage 20-24’C w/ agitation 20-24’C w/ agitation
Shelf-life 5 days 5 days
At risk of TA-GVHD Recipient of BM transplant
Patients w/ hematologic or oncologic disease
Patients w/ congenital immunodeficiency
Recipient of blood from 1st degree relative (direct)
Irradiation Radiation source: 25-35 Gy (Gy: Gray | 1Gy = 100 rads)
a. Cesium (Ce)
b. Cobalt (Co)
Infusion IV fluids:
 a. NSS = the only fluid allowed to start an IV line prior to transfusion
b. D5W (5% dextrose in H2O) = Not allowed to start (hypotonic à hemolysis)
c. Ringer’s lactate =not allowed to start (contains Ca2+ à promote coagulation)
Blood warmers 37’C
áTemp. >42’C = hemolyzed
Filters 1. Clot screen filter (170µm) = to remove gross clots
2. Leukocyte depletion filter
Speed of infusion 15gtts = 1mL
60gtts = 1min
1min = 4mL
1hr = 240mL
Rate = 240mL/hr
1 unit must be completed w/in 4hrs of blood transfusion
Polyagglutination (Hubener-Thomsen-Fridenrich Phenomenon)
Cryptantigens Hidden Ag’s
Covered w/ NANA (N-acetylneuraminic acid) or sialic acid
When NANA is destroyed (by neuraminidase) à Ag’s are exposed
Exposed Ag’s à Agglutination = React w/ tetrasaccharide of Thomas & Winzler (T
receptor)
Acquired Microbially-Associated Polyagglutination
T C. perfringens
V. cholerae
S. pneumoniae
All produce neuraminidase
Th E. coli
Proteus sp.
Produce weaker neuraminidase
Tx Bacterial and viral
Unknown mechanism
Tk “CABS”
C. albicans
A. niger
B. fragilis
S. marcescens
All produce endo/exogalactosidase
Altered precursor substances (Altered: ABH, Le, I, P)
Acquired B phenomenon Bacteria: Deacetylase enzyme
N-acetylgalactosamine --(Deacetylase)--> N-acetyl + galactosamine
Galactosamine = Group B
Remedy: Add acetic anhydride
VA Vienna, Virginia
Microbial fucosidase = âfucose = âH
Acquired Non-Microbially Associated Polyagglutination
Tn (-) β-3-D-galactosyltransferase enzyme needed for the normal structure of T receptor
(Tetrasaccharide of Thomas and Winzler)
Inherited Polyagglutination
Cad áSda
HEMPAS/CDA II áAdult i Ag = âH Ag, âSialic acid
Others HbM – Hyde Park
NOR = Norfolk, Virginia

Tn Cad T Tk
Arachis hypogaea - - + +
Salvia sclarea + - - -
Salvia horminum + + - -
Glycine soja + + + -
 Autologous Donation
Predeposit AD Before anticipated transfusion
Requirements:
*No age limit
*No strict weight requirement (âvol. by 4mL/1 pound below 110)
*Hgb ≥ 11g/dL
*Hct ≥ 33%
*Frequency: every ≥ 3days
Intraoperative AD Collect blood during surgical procedure à reinfused immediately
Immediate preoperative Collect blood à replace patient volume w/ colloid/crystalloid à reinfuse during surgical
hemodilution procedure
Postoperative salvage Drainage tube
Postoperative bleeding à salvaged (saved) à clean and reinfused
Immediate Immune Transfusion Reactions
FNHTR á1’C in temperature
Cause: anti-leukocyte Ab’s (leukoagglutinins)
Prevention: Leukopoor RBCs
Allergic Cause: Donor plasma w/ foreign proteins
Prevention: Washed RBCs
Anaphylactic Afebrile (no fever)
Signs and symptoms occur only after the infusion of only few mL of blood
IgA deficiency w/ anti-IgA antibody
Prevention: Washed RBCs | IgA-deficient donor (rare)
Anaphylactoid Afebrile
Normal IgA w/ anti-IgA to donor IgA
Prevention: Washed RBCs | IgA-deficient donor (rare)
TRALI (NCPE) Cause: Anti-leukocyte Ab’s (leukoagglutinins)
Signs and symptoms resemble respiratory distress
Prevention: Leukopoor RBCs
Hemolytic Bleeding, hypotension, hemoglobinuria, anuria
Delayed Immune Transfusion Reactions
TA-GVHD Cause: T lymphocyte proliferation
Prevention: Irradiated RBCs
PTP Onset of thrombocytopenia
Cause: anti-platelet Ab’s (HPA-1a negative platelets)
Prevention: Therapy
a. Administration of corticosteroid
b. Exchange transfusion
c. IV immunoglobulins
DHTR 7 days
Immediate Nonimmune Transfusion Reactions
TACO Administration of blood w/o equivalent blood loss
Iatrogenic: physician-caused
At risk:
a. Young children
b. Elderly patients
c. Patients w/ cardiac disease
Prevention: Therapy
a. Therapeutic phlebotomy
b. IV diuretics
c. O2 therapy
Bacterial contamination Cause: Endotoxin production by psychrophilic/cryophilic bacteria
Y. enterocolitica (most common)
E. coli
P. aeruginosa
Factors:
a. During phlebotomy
b. During preparation/processing
c. During thawing
Prevention:
a. Sterile technique
b. Visual inspection of unit
→ Blood unit = Brown, purple, hemolysis, clot
→ Plasma = Murky (dark brown) purple, red
PCITR Causes:
- Small bore needle
- Warming blood above 50’C
- Freezing blood w/o cryoprotective agent

Iron Overload (Hemosiderosis) Patients w/ normovolemic anemia
Disease transmission
In utero
Neonatal period
Treatment
Full X-match
Abbreviated X-match
Electronic X-match
Emergency situation
(+) Major X-match
Ratio of serum to cells
Saline for washing
Incubation period
Enhancement media
Anti-A1
Anti-H
Anti-M
Anti-N
Anti-B
- Citrate toxicity
Delayed Nonimmune Transfusion Reactions
Transfusion-dependend patients:
- Aplastic anemia
- Congenital hemolytic anemia
Prevention:
a. Iron-chelating agent = Deferroxamine
b. Neocytes = young RBCs, has longer lifespan
HBV, HCV, HDV, CMV, EBV, HTLV-I and II, HIV, T. pallidum, Plasmodium spp., B.
microti, T. cruzi, T. gondii
Hemolytic Disease of the Newborn
Anemia (á immature RBCs, enlarged spleen & liver = extramedullary hematopoiesis)
Hydrops fetalis = cardiac insufficiency, edema
á Unconjugated bilirubin à Brain à Kernicterus
1. Intrauterine transfusion
- In utero
- Corrects anemia
- X-match: Mother’s serum
2. Exchange transfusion
- Neonatal period
- Removes bilirubin & Ab-coated RBCs
- X-match: Mother’s serum (preferred) or infant’s serum
Cross-Matching
2 hours
Can be shortened to 30 mins
Type/screen + immediate spin
DC/PS = no agglutination/hemolysis
Patient blood type is determined on 2 occasions
Patient blood type is unknown
Group O (Rh-) pRBCs
(-) Alloantibody control
(+) Autoantibody, autocontrol
Routine = 40:1
Det. weak Ab’s = 133:1
pH 7.2-7.4
30-120 mins (majority: 30 mins)
1. Albumin
2. PEG
3. LISS (Incubation pd: 5-15 mins)
Lectin Sources
Dolichos biflorus
Ulex europaeus
Iberis amara
Iberis umbellate
Iberis semperivens
Maclura aurantiaca
Bauhinia variegate
Bauhinia purpura
Bauhinia bonatiana
Bauhinia candicans
Vicia graminea
Bandeirae simplicifolia
 Quality Control
Annually Mercury thermometers
Quarterly (Every 3 months) Speed timer (centrifuge)
Cell washers (speed, timer)
Blood warmers
Monthly Alarm Activation (refrigeration and freezers)
Temperature (refrigerated centrifuge)
Daily Refrigerators and freezers (continuous monitors)
Platelet incubators (enclosed, monitored chambers)
Daily when in use Transfusion service:
- Heating blocks
- Water baths
Donor facility:
- Donor unit agitators
- Scales
- Balances
- Hemoglobinometer
- Microhematocrit centrifuges
Every 4 hours Platelet incubators (ambient temperature storage)
Other Topics
Apheresis/Hemapheresis Whole blood is withdrawn, a desired component separated, and the remainder of the
tube returned to the donor
Intermittent-flow centrifugation 1 venipuncture
(IFC) Blood is withdrawn and reinfused through the same needle
Once the desired component is separated, the remaining components are reinfused to the
donor
Continuous-flow centrifugation 2 venipunctures
(CFC) Withdraw, process and return the blood to the individual simultaneously
Antibody screen Collect new specimen every 3 days in a series of transfusion
Donor & recipient samples Keep for 7 days after transfusion (at 1-6’C)
Compatibility testing Series of tests that ensure safety of transfusion to recipient and donor
1. Review of BB records
2. ABO and Rh
3. Ab screen
4. Cross matching
a. Major X-match = DC/PS
b. Minor X-match = PC/DS
DHTRs Occur 3 to 7 days after transfusion
Virus inactivation in plasma Heating in a liquid state w/ LMW stabilizers
products Heating in lyophilized state
UV irradiation
Y. enterocolitica ááá Bacterial contamination of blood
Density Separates neocytes from mature RBCs
Requirements for blood 1. (-) HbS
products to be transfused to 2. <7 days
infants 3. γ-irradiated
4. (-) CMV
Allogeneic donor blood from Repeat ABO and Rh typing
outside sources
w/in 30 mins Time limit when a unit of blood is removed from 2-8’C and returned back to the
refrigerator
Procedure when HTR occurs 1. Stop transfusion
2. Keep IV line open
3. Notify the physician and BB
Specimens for investigation of 1. Patient pre-transfusion blood sample
HTR 2. Patient post-transfusion blood sample
= PT/PTT/DAT
= Pink: Hgb 0.2 g/L or 20 mg/dL
= Red: Hgb >1 g/L or 100 mg/dL
3. Patient post-transfusion urine
= Bilirubin/Urobilinogen
4. Blood bag
= GS/CS
LISS Glycine/glucose + saline
5-15 mins incubation
ZZAP Cysteine-activated papain
Mixture of DTT and papain
True chimerism
Artificial chimerism
AB cis genotype
Acquired A antigen
Acquired B antigen
HAT medium
Hybridoma cells
Polyspecific AHG reagent
Monospecific AHG reagent
Type 1 precursor substance
Type 2 precursor substance
Agglutination in Gel technology Incompatible
Affinity column technology
(Gamma ReACT)
Wash RBCs
# cryobags
# units to test
Enzyme technique
Elution
Adsorption
Biphasic hemolysin
Perfluorocarbons
Presence of 2 cell population
Ex. Twins
Mixed-field agglutination
After:
-BM transplantation
-Blood transfusion
-Exchange transfusion
-Fetomaternal hemorrhage
Group IV discrepancy
Inheritance of 3 ABO genes (AB/O)
1. Tn activation
2. P. mirabilis
“EPIC”
1. E. coli O86
2. P. vulgaris
3. Intestinal obstruction
4. Carcinoma of the colon and rectum
Hypoxanthine, Aminopterin, Thymidine medium
Culture for hybridoma cells
Plasma cells (mouse) + myeloma cells + PEG
Anti-IgG and anti-C3d
Prepared by conventional technique (rabbits)
Anti-IgG or anti-C3d
Prepared by monoclonal Ab production (mice)
β 1à 3
ABH substance in secretions (glycoproteins)
HAB, Se, Le genes
β 1à4
ABH on RBC (glycoproteins)
HAB genes
Affinity adherence of IgG-sensitized RBCs to immunologically active matrix
2-3x to remove unbound globulins
[Desired level of Factor VIII x volume of plasma (mL)] ÷ 80
# units required ÷ frequency of donors (-) for relevant Ag
1. Papain = papaya
2. Bromelin = pineapple
3. Ficin = figs
4. Trypsin = pig’s stomach
Breaking of bonds between Ag and Ab by:
a. Physical: heating, shaking
b. Chemical: ether, acid
Removal of Ab in serum using appropriate RBC
Cold = attaches to RBCs
Warm = RBC lysis
O2 carrying capacity
RBC substitute
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