Spotlight
TFH cell
Germinal Center
Lymphocyte Ratios
and Successful HIV
Vaccines
Paula Gonzalez-Figueroa,1
Jonathan A. Roco,1 and
Carola G. Vinuesa1,*
Current HIV vaccines are poor
inducers of neutralizing antibodies
(nAbs). A recent study in Cell
Reports used serial ne-needle
aspirates from rhesus macaque
lymph nodes following HIV-1 sur-
face envelope glycoprotein (Env)
trimer immunization, generating a
substantial production of HIV-1
nAbs. A remarkable correlation
was found between antibody titers
and a high frequency and ratio of
germinal center B and T follicular
helper (TFH) lymphocytes.
HIV-1 infection is a major public health
issue, affecting over 36.7 million people
worldwide. Despite huge efforts by the
scientic community over the past three
decades, no effective cure against HIV is
currently available and the protection
afforded by current vaccines is low.
Most successful vaccines to date rely on
the production of nAbs for protection. In
the case of HIV-1, induction of broadly
neutralizing antibodies (bnAbs) can pro-
tect against a diverse spectrum of HIV-1
strains, but is particularly challenging
owing to the high rate of viral mutations
that lead to the emergence of neutraliza-
tion-resistant viruses and to the ability of
HIV-1 to shield conserved epitopes in Env
glycoproteins [1]. HIV-1 bnAbs arise in
viremic controller subjects years after
the initial infection, accumulating a sur-
prisingly high number of somatic muta-
tions as a result of the continuous
evolution of antibodies within germinal such as IL-21, IL-4, and CD40L.
centers (GCs) as viral escape occurs [2]. expansion is also crucial in facilitating the
control of chronic lymphocytic choriome-
To date, nAbs against clinically relevant ningitis viral infection in mice as well as in
HIV-1 strains (Tier-2 and Tier-3) have not supporting the production of high-afnity
been detected in HIV-1 vaccine trials, and antibodies in SIV-infected RMs. Never-
the mechanisms needed to induce them theless, in the case of chronic HIV/SIV
are poorly understood. To improve the infection, TFH cell expansion has been
quality of the antibodies elicited, one step consistently found to correlate with
forward has been the design of Env trimers increased GC B cell numbers but to neg-
that adopt a native-like conformation atively impact on the quality of HIV-1 anti-
which exposes the key neutralizing epito- body responses [5,6]. It has been
pes to B cells and hides the immunodo- suggested that excess TFH cells corrupt
minant non-neutralizing epitopes [1]. B cell selection owing to the low selection
threshold imposed to GC B cells [7].
A study by Havenar-Daughton and col-
leagues reports the generation of signi- This study suggests that there are char-
cant titers of autologous Tier-2 nAbs in acteristics of the GC response beyond
rhesus macaques (RMs) following estimation of cell frequencies that may
sequential HIV-1 Env trimer immunization be important predictors of the quality of
[3]. Using cutting-edge techniques, the antibody responses. The authors showed
group identied cellular biomarkers in that TFH cells from top neutralizer individ-
draining lymph nodes (LNs) that may pre- uals exhibit increased expression of mol-
dict nAb responses in vaccinated RMs. ecules associated with B cell help
After vaccination, high frequencies of GC (CD40L, IL-21, ICOS), with a high ratio
B and TFH cells as well as a high GC B to of GC B cells to TFH cells. Indeed, a high
TFH cell ratio emerged as good indicators GC B:TFH cell ratio suggests sustained B
of nAb responses (Figure 1). The same cell competition for TFH cell help, which
group previously showed that high num- has been deemed important for afnity-
bers of circulating TFH cells correlated based selection [7]. Therefore, measuring
with bnAb responses in HIV-infected indi- GC B:TFH cell ratios rather than individual
viduals [4]. subset frequencies may constitute a more
accurate assessment of effective anti-
This is the rst study to show that HIV-1 body responses.
protein vaccines are able to induce strong
GC responses in primates. Fine-needle Investigating the contribution of follicular
aspirate (FNA) collection was validated regulatory T (TFR) cells to this response will
as a robust technique to study and moni- now be relevant. TFR cells can regulate the
tor GC cell changes in LNs. Moreover, the magnitude and output of the GC reaction
work demonstrates that FNAs allow lon- and the proportions of GC TFH and/or B
gitudinal analyses in vaccinated subjects cells. Furthermore, TFR cells have been
over multiple time-points with no disrup- recently shown to expand after HIV-1/
tion of GC processes. SIV infection [8] and could represent
one of the missing pieces to help to
TFH cells are particularly susceptible to explain the presence of dysfunctional
HIV-1 or simian immunodeciency virus TFH and CD8+[74_TD$IF] T cells in GCs during
(SIV) infection owing at least in part to the chronic viral infection.
high expression of the HIV-1 coreceptor
CCR5 by TFH cell precursors. TFH cells are This study also reports the highest titers
crucial for afnity-based B cell selection to date against a Tier-2 HIV-1 virus in non-
and differentiation through the production human primates. How did the authors
of cytokines and costimulatory molecules manage to achieve such high nAb
Trends in Molecular Medicine, February 2017, Vol. 23, No. 2 95
 these antibodies in their ability to neutral-
ize different HIV-1 strains.

Despite the dramatic increase in antibody

titers compared to previous vaccination
strategies, approximately 25% of vacci-
nated RMs still failed to produce nAbs.
Further improvements may come from
efforts aiming to mimic more closely the
production of naturally occurring bnAbs.
Recent studies have shown that the
induction of bnAbs against HIV-1 could
be guided in animal models by reverse
engineering the priming immunogen in a
multiple immunization regime [10]. This
approach is supported by the fact that
the inferred germline versions of known
bnAbs show negligible afnity for the
native Env structure and require priming
Figure 1. Increased GC Lymphocyte Responses and B:TFH Cell Ratios Mark Neutralizing Anti-
with a mutated version to be activated.
body (nAb) Production Against HIV-1. Different outcomes of nAb production following multiple immu-
nizations with an engineered HIV-1 Env trimer in rhesus macaques are shown. Lymph nodes from vaccinated Sequential challenges using native-like
animals are sequentially sampled by ne-needle aspirates (FNA) to study GC responses. Top-neutralizing trimers with decreasing number of muta-
animals are characterized by greater TFH and GC numbers and a higher GC B:TFH cell ratio. The optimal tions have been shown to direct the afn-
frequency of GC B and TFH cells in these animals may represent a powerful biomarker to predict the production
of nAb against HIV-1. Abbreviations: Env, HIV-1 surface envelope glycoprotein; GC, germinal center; TFH, T
ity maturation of the germline form of
follicular helper cell. known bnAbs. Adding this strategy to
the immunization regime used in this
responses? This could be the result of a a non-neutralizing immunodominant epi- study may help to close the response
combination of several factors. First, they tope. However, the latter is unlikely to gap in HIV-1 vaccine trials and bring us
took advantage of a sequential immuni- explain the success of the vaccination closer to a much needed end of the HIV
zation approach to sustain the GC reac- strategy given that the modied trimer epidemic.
tion and enhance memory B cell recall was not able to reduce the immunogenic-
responses. ity of the V3 loop. No differences were Acknowledgments
observed between the use of two different C.G.V. is funded by the National Health and Medical
Second, the protocol involved injecting adjuvants, one of which, Iscomatrix, has Research Council (NHMRC) of Australia.
Env at two subcutaneous sites, which been previously used to immunize RMs
1
the authors predicted would engage with Env trimer, and it did not elicit the Department of Immunology and Infectious Disease, John
Curtin School of Medical Research, Australian National
more B cells and T cells. Indeed, through robust immune response observed in this University, Canberra, Australia
B cell receptor (BCR) sequencing they study [9].
showed that different and non-overlap- *Correspondence:
carola.vinuesa@anu.edu.au (C.G. Vinuesa).
ping clones could be found at the two Although it is not possible to conclude
http://dx.doi.org/10.1016/j.molmed.2016.12.009
sites. The cells recruited into the GC reac- which of the various factors drove the high
tion were likely to be Env-specic B cells, production of nAb titers because they References
and phylogenetic analysis suggested were not evaluated independently, it is 1. Haynes, B.F. et al. (2016) HIVhost Interactions:
implications for vaccine design. Cell Host Microbe 19,
that these cells could have undergone likely that multiple doses at more than 292303
afnity maturation. Additional studies are one site may have been key for the robust 2. van Gils, M.J. and Sanders, R.W. (2013) Broadly neutral-
izing antibodies against HIV-1: templates for a vaccine.
needed to identify those B cell clones that and persistent TFH and GC reaction pre- Virology 435, 4656
were specically induced by neutralizing dicted to underpin the nAb response. 3. Havenar-Daughton, C. (2016) Direct probing of germinal
epitopes. center responses reveals immunological features and bot-
tlenecks for neutralizing antibody responses to HIV Env
A crucial next step will be to determine the trimer. Cell Rep. 17, 21952209
Third, the authors used a modied Env potential of these nAbs to protect from 4. Locci, M. et al. (2013) Human circulating PD-
1+CXCR3 CXCR5+ memory Tfh cells are highly functional
trimer predicted to increase its stability HIV-1 infection in vivo. It will also be and correlate with broadly neutralizing HIV antibody
and reduce the exposure of the V3 loop, important to establish the breadth of responses. Immunity 39, 758769

96 Trends in Molecular Medicine, February 2017, Vol. 23, No. 2

5. Onabajo, O.O. and Mattapallil, J.J. (2013) Expansion or
depletion of T follicular helper cells during HIV infection:
consequences for B cell responses. Curr. HIV Res. 11,
595600
6. Petrovas, C. et al. (2012) CD4 T follicular helper cell dynamics
during SIV infection. J. Clin. Invest. 122, 32813294
7. Pratama, A. and Vinuesa, C.G. (2014) Control of TFH
cell numbers: why and how? Immunol. Cell Biol. 92,
4048
8. Miles, B. et al. (2015) Follicular regulatory T cells impair
follicular T helper cells in HIV and SIV infection. Nat. Com-
mun. 6, 8608
9. Sanders, R.W. et al. (2015) HIV-1 neutralizing antibodies
induced by native-like envelope trimers. Science 349,
aac4223
10. Escolano, A. et al. (2016) Sequential immunization elicits
broadly neutralizing anti-HIV-1 antibodies in Ig knockin
mice. Cell 166, 14451458
Spotlight
Metabolic
Syndrome: One
Speckled Stone Kills
a Flock of Birds?
Ylva Bonde1 and
Bo Angelin1,*
Effectively treating metabolic
syndrome and its progression to
type 2 diabetes, steatohepatitis
and cardiovascular disease remain
a major clinical challenge. The use
of a novel engineered molecule
that combines thyroid hormone
and glucagon to target liver and
adipose tissue might provide a
new magic bullet with exciting
future prospects.
Treating the increasingly prevalent inci-
dence of a complex metabolic phenotype
with the aim to retard progression into
type 2 diabetes, steatohepatitis, athero-
sclerosis and/or cardiovascular disease
remains a major obstacle to many physi-
cians. Such patients can suffer from
abdominal obesity, insulin resistance,
dyslipidemia, hypertension, and fatty liver
disease [1]. While there is good evidence
for the benet of treating each of these
traits actively, current practice generally
involves multidrug therapy with risks of
unfavorable drug interactions, unwanted
side effects, or simply an inability to reach
desirable treatment goals. There is there-
fore great interest in the development
of novel therapies to efciently target
several common pathways of metabolism
and reinstitute homeostasis. Many such
efforts are based on our increased
knowledge of how natural hormones or
nutrients may exert integrated control of
metabolism, as for instance through their
actions as ligands for nuclear receptors.
Thyroid hormone (TH) has long been rec-
ognized as a potent regulator of multiple
metabolic pathways by interacting with
TH receptors (TRs) in various tissues such
as liver and adipose tissue [2]. Lipoprotein
metabolism in strongly inuenced by TH,
and dyslipidemia is common in thyroid
disorders. TH stimulates processes that
contribute to the elimination of cholesterol
from the body, including its conversion
into bile acids. TH also reduces intestinal
absorption of cholesterol and promotes
fat oxidation and energy expenditure,
resulting in reduced body weight and
improved glucose metabolism. However,
negative effects on heart, muscle, and
bone have been found to occur following
administration of TH, thus precluding its
therapeutic use [2]. Renement of more
selective thyromimetics has proceeded
essentially along two paths: either
through elaboration of analogs with
higher afnity to the TR-b isoform mainly
expressed in the liver and regulating cho-
lesterol metabolism or through develop-
ment of analogs that are selectively taken
up by or activated within hepatocytes.
Such compounds lower plasma low-den-
sity lipoprotein (LDL) and triglycerides
(TGs) without inuencing heart or muscle
function, or perturbing the hypothalamic
pituitarythyroid axis [2]. However, possi-
bly mainly due to the lack of TH effects on
adipose tissue, these agents do not sub-
stantially inuence body weight. Despite
promising results in clinical trials [3,4],
further development of the leading liver-
Trends in Molecular Medicine, February 2017, Vol. 23, No. 2
selective thyromimetic, eprotirome, was
terminated in 2012 following observations
of cartilage damage in long-term toxicity
studies in dogs [4].
Researchers at the Helmholtz Diabetes
Center in Munich now present extensive
preclinical data regarding treatment with a
novel compound that may open exciting
possibilities for future clinical develop-
ment [5]. The novel study, published in
Cell, is conceptually based on their previ-
ous ground-breaking work that demon-
strates how unimolecular combination
therapeutics such as the administration
of conjugate molecules of glucagon and
glucagon-like peptide-1 can lead to
improved pharmacological activity [6,7].
They now report in several murine disease
models that by coupling triiodothyronine
(T3) to glucagon, it is possible to selec-
tively target TH to the liver and adipose
tissue, and consequently, to simulta-
neously obtain synergistic effects from
both hormones to normalize metabolic
disturbances [5]. The engineered T3/
glucagon conjugate is actively taken up
by hepatocytes and to some extent
by white adipocytes via the glucagon
receptor. The internalized T3 can then
activate TR-regulated metabolic path-
ways. Through the hepatic b receptors,
bile acid synthesis is stimulated, and the
expression of LDL receptors (LDLRs) and
the high-density lipoprotein (HDL) recep-
tor, scavenger receptor class B member
1 (SRB1), is increased, resulting in a
marked lowering of plasma cholesterol
(Figure 1). Also, the hepatic production
of TG is reduced, leading to lower levels
in both plasma and liver. In addition, the
conjugate reduces body weight in ani-
mals fed a high-fat diet by increasing
energy expenditure [5].
The tissue selectivity of the T3/glucagon
compound relative to the administration
of pure T3 can be adequately explained
by the distribution of glucagon receptors,
which critically decides the selective
uptake of the conjugate compound. Sim-
ilarly, the pronounced effects on bile acid
97