Coadministration of Polyinosinic: Polycytidylic Acid and Immunostimulatory Complexes Modifies Antigen Processing in Dendritic Cell Subsets and Enhances HIV Gag-Specific T Cell Immunity

Article (PDF Available)inThe Journal of Immunology 191(10) · October 2013with24 Reads
DOI: 10.4049/jimmunol.1301730 · Source: PubMed
Abstract
Currently approved adjuvants induce protective Ab responses but are more limited for generating cellular immunity. In this study, we assessed the effect of combining two adjuvants with distinct mechanisms of action on their ability to prime T cells: the TLR3 ligand, polyinosinic:polycytidylic acid (poly I:C), and immunostimulatory complexes (ISCOMs). Each adjuvant was administered alone or together with HIV Gag protein (Gag), and the magnitude, quality, and phenotype of Gag-specific T cell responses were assessed. For CD8 T cells, all adjuvants induced a comparable response magnitude, but combining poly I:C with ISCOMs induced a high frequency of CD127(+), IL-2-producing cells with decreased expression of Tbet compared with either adjuvant alone. For CD4 T cells, combining poly I:C and ISCOMs increased the frequency of multifunctional cells, producing IFN-γ, IL-2, and TNF, and the total magnitude of the response compared with either adjuvant alone. CD8 or CD4 T cell responses induced by both adjuvants mediated protection against Gag-expressing Listeria monocytogenes or vaccinia viral infections. Poly I:C and ISCOMs can alter Ag uptake and/or processing, and we therefore used fluorescently labeled HIV Gag and DQ-OVA to assess these mechanisms, respectively, in multiple dendritic cell subsets. Poly I:C promoted uptake and retention of Ag, whereas ISCOMs enhanced Ag degradation. Combining poly I:C and ISCOMs caused substantial death of dendritic cells but persistence of degraded Ag. These data illustrate how combining adjuvants, such as poly I:C and ISCOMs, that modulate Ag processing and have potent innate activity, can enhance the magnitude, quality, and phenotype of T cell immunity.
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Immunity
and Enhances HIV Gag-Specific T Cell
Antigen Processing in Dendritic Cell Subsets
Immunostimulatory Complexes Modifies
Polyinosinic:Polycytidylic Acid and
Coadministration of
Johnson, Barbara J. Flynn, Karin Loré and Robert A. Seder
A. Darrah, Ross W. B. Lindsay, Sonia T. Hegde, Teresa R.
Kylie M. Quinn, Ayako Yamamoto, Andreia Costa, Patricia
http://www.jimmunol.org/content/191/10/5085
doi: 10.4049/jimmunol.1301730
October 2013;
2013; 191:5085-5096; Prepublished online 2J Immunol
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The Journal of Immunology
Coadministration of Polyinosinic:Polycytidylic Acid and
Immunostimulatory Complexes Modifies Antigen Processing
in Dendritic Cell Subsets and Enhances HIV Gag-Specific
T Cell Immunity
Kylie M. Quinn,* Ayako Yamamoto,* Andreia Costa,*
,1
Patricia A. Darrah,*
Ross W. B. Lindsay,*
,2
Sonia T. Hegde,*
,3
Teresa R. Johnson,*
,4
Barbara J. Flynn,*
Karin Lore
´
,*
,†
and Robert A. Seder*
Currently approved adjuvants induce protective Ab responses but are more limited for generating cellular immunity. In this study,
we assessed the effect of combining two adjuvants with distinct mechanisms of action on their ability to prime T cells: the TLR3
ligand, polyinosinic:polycytidylic acid (poly I:C), and immunostimulatory complexes (ISCOMs). Each adjuvant was administered
alone or together with HIV Gag protein (Gag), and the magnitude, quality, and phenotype of Gag-specific T cell responses were
assessed. For CD8 T cells, all adjuvants induced a comparable response magnitude, but combining poly I:C with ISCOMs induced
a high frequency of CD127
+
, IL-2–producing cells with decreased expression of Tbet compared with either adjuvant alone. For
CD4 T cells, combining poly I:C and ISCOMs increased the frequency of multifunctional cells, producing IFN-g, IL-2, and TNF,
and the total magnitude of the response compared with either adjuvant alone. CD8 or CD4 T cell responses induced by both
adjuvants mediated protection against Gag-expressing Listeria monocytogenes or vaccinia viral infections. Poly I:C and ISCOMs
can alter Ag uptake and/or processing, and we therefore used fluorescently labeled HIV Gag and DQ-OVA to assess these
mechanisms, respectively, in multiple dendritic cell subsets. Poly I:C promoted uptake and retention of Ag, whereas ISCOMs
enhanced Ag degradation. Combining poly I:C and ISCOMs caused substantial death of dendritic cells but persistence of
degraded Ag. These data illustrate how combining adjuvants, such as poly I:C and ISCOMs, that modulate Ag processing and
have potent innate activity, can enhance the magnitude, quality, and phenotype of T cell immunity. The Journal of Immunology,
2013, 191: 5085–5096.
P
reventive vaccination against HIV, malaria, and tubercu-
losis will require induction of potent Ab responses, T cell
responses, or both for optimal protection (1–4). Because
humoral and cellular immune responses can wane following vac-
cination, continued boosting may be required to maintain responses
above a threshold necessary to mediate protection. Protein-based
vaccines given with adjuvants are one approach that can be used in
combination with other vaccine platforms for priming and/or
boosting adaptive immunity. Currently approved clinical adju-
vant formulations include alum and oil/water emulsions, which
elicit protective humoral immunity, but are far less potent for
inducing CD4/Th1 or CD8 T cell immunity [revie wed in (5)]. Im-
proving cellular immunity with protein-based vaccination will re-
quire adjuvants that elicit potent innate cytokines conducive to
induction of cellular responses and efficient Ag presentation.
Polyinosinic:polycytidylic acid (poly I:C) and immunostimula-
tory complexes (ISCOMs) are two adjuvants that show promise in
preclinical studies and early clinical trials for induction of both
Ab and T cell responses (6–9).
Poly I:C is a synthetic dsRNA analog and a ligand for multi-
ple pathogen recognition receptors, including TLR3, melanoma dif-
ferentiation-associated protein 5, retinoic acid-inducible gene 1, and
dsRNA-dependent protein kinase R (10–14). Expression of TLR3
is endosomal and found predominantly in C D8a
+
dendritic cells
(DCs) or langerin
+
dermal DCs (dDCs) (15, 16), whereas melanoma
differentiation-associated protein 5, retinoic acid-inducible gene 1,
and protein kinase R localize to the cytosol and are more broadly
expressed on APCs and nonhematopoietic stromal cells (6, 17,
18). Poly I:C stimulates rapid production of IL-6, IL-10, IL-12
p40,MCP-1,TNF,typeIIFN,andIFN-g, resulting in significant
DC and NK cell activation (6, 19). When coadministered with
protein Ag, poly I:C potently primes CD4/Th1 cell and Ab responses
(6, 7, 20) and promotes cross-presentation of Ag to CD8 T cells by
DCs through TLR3 signaling (21).
ISCOM particles are cage-like structures that assemble from
cholesterol, phospholipids, and saponins [re vie wed in (22)]. ISCOMs
*Vaccine Research Center, National Institute of Allergy and Infectious Diseases,
National Institutes of Health, Bethesda, MD 20892; and
Center for Infectious Med-
icine, Department of Medicine, Karolinska Institutet, Stockholm, SE-171 77, Sweden
1
Current address: Fred Hu tchinson Cancer Research Center, Seattle, WA.
2
Current address: International AIDS Vaccine Initiative, Brooklyn, NY.
3
Current address: Centers for Disease Control and Prevention, Atlanta, GA.
4
Current address: GenVec, Gaithersburg, MD.
Received for publication June 28, 2013. Accepted for publication September 8, 2013.
This work was supported in part by a grant from the Foundation for t he National
Institutes of Health with support from Collaboration for AIDS Vaccine Discovery
Award OPP1039775 from the Bill and Melinda Gates Foundation.
Address correspondence and reprint requests to Dr. Robert A. Seder, Cellular Immu-
nology Section, Vaccine Research Center, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, 40 Convent Drive MSC 3025, Building 40,
Room 3512, Bethesda, MD 20892. E-mail address: rseder@mail.nih.gov
The online version of this article contains supplemental material.
Abbreviations used in this article: AF, Alexa Fluor; DC, dendritic cell; dDC, dermal
DC; dLN, draining lymph node; ICS, intracellular cytokine staining; ISCOM, immu-
nostimulatory complex; KLRG1, killer-cell lectin–like receptor subfamily G1; MFI,
median fluorescent intensity; pDC, plasmacytoid DC; poly I:C, polyinosinic:polycy-
tidylic acid; WT, wild-type.
www.jimmunol.org/cgi/doi/10.4049/jimmunol.1301730
by guest on January 9, 2016http://www.jimmunol.org/Downloaded from
can enhance Ag delivery to APCs when Ag is incorporated into the
particle, but ISCOMs do not function solely as deliv ery vehicles,
because certain fractions of saponin possess intrinsic adjuv ant ac-
tivity (23). ISCOMs have been shown to induce caspase-dependent
cleavage of IL-1b and robust serum production of IL-5, IL-6, GM-
CSF, and IL-12 p40 (24, 25). As a result, ISCOMs prime potent
long-lived Ab responses with a balanced CD4 Th1/Th2 T cell re-
sponse (26) and low-level induction of CTLs. I SCOMs lead to
cross-presentation most likely as a result of disruption of the in-
tegrity of phagolysosomes after endocytosis, which could permit
access of Ag to the cytosol (27, 28). Cross-presentation with
ISCOMs in vitro is most efficient with monocyte-derived DCs (28),
although ex vivo CD8a
+
DCs are responsible for the majority of Ag
presentation to CD8 T cells (25).
A combination of poly I:C and ISCOMs could potentiate the
effect of each adjuvant by activating distinct, but complimentary
innate signaling and Ag-processing pathways. Prior studies using
combined ligands for distinct TLRs have demonstrated enhanced
innate or adaptive immunity in vitro (29) and in vivo (30). Poly I:C
has been used in combination with particulate delivery systems,
such as liposomes, and combined with other TLR agonists, such as
CpG, to enhance innate signaling and priming of T cells (31, 32).
ISCOMs have also been used in combination with CpG, which
enhanced cross-priming of tumor Ag (33) and induced robust HIV
Env-specific humoral immunity (34). However, a combination of
poly I:C with ISCOMs has not been evaluated.
For this study, we hypothesized that combining poly I:C and
ISCOMs would result in more potent T cell immunity than either
adjuvant alone. We show that combining the adjuvants increased
CD4/Th1 cell responses and enhanced the qualitative and phe-
notypic profile of CD8 T cell responses by increasing expression of
CD127 and IL-2. Combining poly I:C with ISCOMs also resulted
in rapid initial degradation but prolonged retention of Ag, which
may represent a mechanism contributing to the observed effects on
T cells. Overall, the