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. 2008 Feb 19;105(7):2574-9.
doi: 10.1073/pnas.0711976105. Epub 2008 Feb 6.

The microbial mimic poly IC induces durable and protective CD4+ T cell immunity together with a dendritic cell targeted vaccine

Christine Trumpfheller  1 , Marina Caskey, Godwin Nchinda, Maria Paula Longhi, Olga Mizenina, Yaoxing Huang, Sarah J Schlesinger, Marco Colonna, Ralph M Steinman
Affiliations

The microbial mimic poly IC induces durable and protective CD4+ T cell immunity together with a dendritic cell targeted vaccine

Christine Trumpfheller et al. Proc Natl Acad Sci U S A. .

Abstract

CD4(+) Th1 type immunity is implicated in resistance to global infectious diseases. To improve the efficacy of T cell immunity induced by human immunodeficiency virus (HIV) vaccines, we are developing a protein-based approach that directly harnesses the function of dendritic cells (DCs) in intact lymphoid tissues. Vaccine proteins are selectively delivered to DCs by antibodies to DEC-205/CD205, a receptor for antigen presentation. We find that polyriboinosinic:polyribocytidylic acid (poly IC) independently serves as an adjuvant to allow a DC-targeted protein to induce protective CD4(+) T cell responses at a mucosal surface, the airway. After two doses of DEC-targeted, HIV gag p24 along with poly IC, responder CD4(+) T cells have qualitative features that have been correlated with protective function. The T cells simultaneously make IFN-gamma, tumor necrosis factor (TNF)-alpha, and IL-2, and in high amounts for prolonged periods. The T cells also proliferate and continue to secrete IFN-gamma in response to HIV gag p24. The adjuvant role of poly IC requires Toll-like receptor (TLR) 3 and melanoma differentiation-associated gene-5 (MDA5) receptors, but its analog poly IC(12)U requires only TLR3. We suggest that poly IC be tested as an adjuvant with DC-targeted vaccines to induce numerous multifunctional CD4(+) Th1 cells with proliferative capacity.

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Conflict of interest statement

Conflict of interest statement: R.M.S. is a consultant to Celldex, which is developing human DEC-205-based vaccines. All other authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Properties of HIV gag-specific CD4+ T cells after prime-boost immunization with α-DEC-p24 and poly IC. C57BL/6 mice were injected s.c. with 5 μg of α-DEC-p24 and 2, 10, or 50 μg of poly IC or PBS, and boosted with the same conditions 6 wk later. One other group received 5 μg of α-DEC-p24 and 50 μg of poly IC once at the time of the boost. Two weeks later, HIV gag-specific, CD3+ CD4+, splenic T cells were analyzed for IFN-γ-, IL-2-, and TNF-α-secretion (A), percentage of CD4+ T cells expressing each of seven possible combinations of IFN-γ, TNF-α, and IL-2 (B), and IFN-γ, TNF-α, and IL-2 MFIs of gag-specific cells expressing 3+, 2+, or 1+ cytokines after prime-boost with α-DEC-p24 plus 50 μg of poly IC (C). Data are representative of two experiments with two mice pooled in each.
Fig. 2.
Fig. 2.
Prime-boost immunization with α-DEC-p24 and poly IC induces long-lived IFN-γ-secreting and proliferating CD4+ T cells. (A) As in Fig. 1, but 2 wk after boosting, bulk splenocytes were CFSE-labeled and stimulated with α-CD28 and gag p24 peptide mix, control gag p17 peptide mix, or α-CD3 for 3 days, whereupon the cells were restimulated for 6 h with HIV gag p24 peptides to detect IFN-γ in proliferated CFSElow, CD3+CD4+ T cells. Data are the percentage of IFN-γ+ or IFN-γ− CD3+CD4+ proliferating T cells (Top Left and Bottom Left, respectively) from one of two similar experiments with two mice pooled in each. (B) As in A, but mean ± SD of the frequencies of IFN-γ-producing and proliferating CD3+CD4+ T cells in three experiments in BALB/c or C57BL/6 mice immunized twice with the indicated vaccines at a 6-wk interval, 2 and 7 wk after boost (except control Ig-p24; n = 2 in C57BL/6 and n = 1 in BALB/c mice).
Fig. 3.
Fig. 3.
α-DEC-nef and poly IC induces nef-specific CD4+ T cell immunity. BALB/c mice were injected twice i.p. with 5 μg of α-DEC-nef or unconjugated α-DEC mAb and 50 μg of poly IC at a 6-wk interval. Nef-specific T cells were analyzed a week after boosting. (A) IFN-γ in spleen CD3+CD4+ T cells in response to HIV nef peptide mix, different nef peptide pools, or nonreactive peptide mix. (B) CFSE-labeled bulk splenocytes were stimulated with α-CD28 and nef peptide mix, nonreactive peptide mix or α-CD3 for 3 days, whereupon the cells were restimulated with HIV nef peptides to detect IFN-γ in proliferated CFSElow, CD3+CD4+ T cells. Shown is the percentage of IFN-γ+ or IFN-γ− CD3+CD4+ proliferating T cells (Upper Left and Lower Left, respectively) from one of three experiments with two mice pooled in each.
Fig. 4.
Fig. 4.
Prime-boost immunization with 5 μg of α-DEC-gag and 50 μg of poly IC provides protective immunity to airway challenge with vaccinia-gag virus. (A) Average weight loss (Left) and lung virus titers (Right) as a mean ± SD after challenge from six experiments, three each in BALB/c and C57BL/6 mice (except control Ig-p24; n = 2 in BALB/c). Mice were primed and boosted at a 6-wk interval with the indicated vaccines. Another group was immunized with α-DEC-p24 and poly IC at the time of the boost only. Six to eight weeks after boost, mice were challenged i.n. with 5 × 104 PFU vaccinia-gag. (B) As in A, but C57BL/6, DEC-205−/−, or TLR3−/− mice were immunized with α-DEC-p41 and challenged with vaccinia-gag. Data are one of two similar experiments. Data are average body weight (Left) or vaccinia plaque-forming titers in lung (Right) as mean ± SD (n = 5).
Fig. 5.
Fig. 5.
Poly IC12U acts as an adjuvant for CD4+ T cell immunity to α-DEC-p24 vaccine in a TLR3-dependent manner. (A) CxB6 F1 mice were injected i.p. with 5 μg of α-DEC-p24 plus graded doses of poly IC (InVivoGen) or poly IC12U (Ampligen) or PBS, and boosted with the same conditions 6 wk later. The percentage of IFN-γ-producing and proliferating CD3+CD4+ T cells in response to HIV gag p17 or p24 mix 1 wk after boost are shown as mean ± SD (n = 4 mice). (B) IFN-γ-secretion in response to HIV gag p24 peptides by CD4+ splenocytes in wild-type, TLR3−/−, and MDA5−/− mice immunized with two doses of α-DEC-p24 plus 50 μg of poly IC (InVivoGen) or 250 μg of poly IC12U (Ampligen). Data are one of two similar experiments with two mice in each.
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