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. 2011 Jan;92(Pt 1):128-40.
doi: 10.1099/vir.0.023242-0.

Induction of Mucosal and Systemic Antibody and T-cell Responses Following Prime-Boost Immunization With Novel Adjuvanted Human Immunodeficiency virus-1-vaccine Formulations

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Free PMC article

Induction of Mucosal and Systemic Antibody and T-cell Responses Following Prime-Boost Immunization With Novel Adjuvanted Human Immunodeficiency virus-1-vaccine Formulations

Anthony D Cristillo et al. J Gen Virol. .
Free PMC article

Abstract

As sexual transmission of human immunodeficiency virus-1 (HIV-1) occurs via the mucosa, an ideal HIV-1 vaccine should induce both mucosal and systemic immunity. We therefore sought to evaluate the induction of mucosal responses using a DNA env prime-gp120 protein boost approach in which sequential nasal and parenteral protein administration was performed with two novel carbohydrate-based adjuvants. These adjuvants, Advax-M and Advax-P, were specifically designed for mucosal and systemic immune enhancement, respectively. Murine intranasal immunization with gp120/Advax-M adjuvant elicited gp120-specific IgA in serum and mucosal secretions that was markedly enhanced by DNA priming. Boosting of DNA-primed mice with gp120/Advax-M and gp120/Advax-P by sequential intranasal and intramuscular immunization, or vice versa, elicited persistent mucosal gp120-specific IgA, systemic IgG and memory T- and B-cell responses. Induction of homologous, but not heterologous, neutralizing activity was noted in the sera of all immunized groups. While confirmation of efficacy is required in challenge studies using non-human primates, these results suggest that the combination of DNA priming with sequential nasal and parenteral protein boosting, with appropriate mucosal and systemic adjuvants, could generate strong mucosal and systemic immunity and may block HIV-1 mucosal transmission and infection.

Figures

Fig. 1.
Fig. 1.
Murine immunization schedule. (a) Mice (n=5) were immunized with recombinant gp120 (25 μg) via the IM route for Advax-P1 (1 mg) or via the IN route for Advax-M (2 μg) at weeks 0, 2 and 5. (b) Alternatively, mice (n=5) were immunized with DNA (100 μg) at weeks 0, 2 and 4, and with adjuvanted gp120 protein at weeks 9 and 11. (c) Mice (n=10) were immunized intramuscularly with DNA (100 μg) at weeks 0, 2 and 4 and boosted with adjuvanted gp120 (25 μg) for either Advax-M or Advax-P1 via the IN or IM routes, respectively, at weeks 9 and 11. Combination delivery strategies were tested in which gp120 was administered by the IM route for Advax-P1 at week 9 and by the IN route for Advax-M at week 11 or vice versa.
Fig. 2.
Fig. 2.
Generation of anti-gp120 IgA antibodies in serum and mucosal compartments of mice immunized with DNA and adjuvanted protein. At 2 weeks post-protein immunization, anti-gp120 IgA responses, assayed by ELISA, were determined for serum (upper panels), vaginal wash samples (middle panels) and saliva (lower panels) following protein (a), and DNA prime–protein boost (b), immunization. Mean serum titres±sem values are shown. For vaginal-wash samples (1 : 2 dilution) and saliva (1 : 5 dilution), mean A450 measurements±sem are shown for the diluted samples.
Fig. 3.
Fig. 3.
T-cell responses in splenocytes of mice immunized with DNA and adjuvanted protein. Secreted Th1 (TNF-α, IFN-γ and IL-2) and Th2 (IL-5 and IL-4) cytokines were quantified, by CBA, following a 24 h ex vivo stimulation of splenocytes with 1 μg  Env peptide pool ml−1, from protein-immunized (a) and DNA prime–protein boosted (b) mice. Mean cytokine responses for each group±sem values are shown.
Fig. 4.
Fig. 4.
Durability of anti-gp120 IgG and serum and mucosal IgA responses following immunization of mice via parenteral, mucosal and combination routes. Anti-gp120 IgG (a, left panel) and IgA (a, right panel) responses, assayed by ELISA, were determined for serum samples of DNA prime–protein boosted mice at weeks 13, 15, 18, 22 and 34 of the study. IgA was also measured at weeks 13 and 34 of the study for saliva (b, 1 : 5 dilution) and vaginal-wash samples (c, 1 : 2 dilution). ELISA titres and A450 are reported.
Fig. 5.
Fig. 5.
Durability of gp120-specific Th1 and Th2 cytokine recall responses. Cellular responses were measured, following ex vivo Env peptide stimulation, by IFN-γ ELISPOT and CBA assays at 2 (left panels) and 23 weeks (right panels) post-protein boost. IFN-γ production, measured by ELISPOT assay (a), and secreted TNF-α (b), IFN-γ (c), IL-2 (d) IL-4 (e) and IL-5 (f) levels measured by CBA, are shown.
Fig. 6.
Fig. 6.
Generation of B- and T-cell memory following immunization of mice via parenteral, mucosal and combination routes. At 23 weeks post-final immunization, splenocytes (1×105, 2×104 and 4×103) were stimulated for 7 days with a ConA supernatant mixture with mitomycin-treated splenocyte feeder cells (a). Alternatively, splenocytes (1×105, 2×104 and 4×103) were stimulated for 7 days with 4 mg CpG-ODN 2006 ml−1 with mitomycin-treated feeder cells (b). gp120-specific antibodies were measured in the media by ELISA, and A450 readings are shown. Alternatively, splenocytes (1.25×106 cells ml−1) were stimulated for 24 h with 1 μg Env peptide pool ml−1 at 37 °C, 5 % CO2 in the presence of 1 μg GolgiPlug (BD Bio Sciences) ml−1 for the final 6 h of stimulation. CD8 and CD4 effector memory (CD44hiCD62L) (c) and central memory (CD44hiCD62L+) (d) T-cell responses, as measured by Th1 cytokine production.

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References

    1. Ahmed, R. K., Biberfeld, G. & Thorstensson, R. (2005). Innate immunity in experimental SIV infection and vaccination. Mol Immunol 42, 251–258. - PubMed
    1. Alving, C. R. & Rao, M. (2008). Lipid A and liposomes containing lipid A as antigens and adjuvants. Vaccine 26, 3036–3045. - PubMed
    1. Asahi-Ozaki, Y., Yoshikawa, T., Iwakura, Y., Suzuki, Y., Tamura, S., Kurata, T. & Sata, T. (2004). Secretory IgA antibodies provide cross-protection against infection with different strains of influenza B virus. J Med Virol 74, 328–335. - PubMed
    1. Barnett, S. W., Srivastava, I. K., Kan, E., Zhou, F., Goodsell, A., Cristillo, A. D., Ferrai, M. G., Weiss, D. E., Letvin, N. L. & other authors (2008). Protection of macaques against vaginal SHIV challenge by systemic or mucosal and systemic vaccinations with HIV-envelope. AIDS 22, 339–348. - PubMed
    1. Barone, F., Patel, P., Sanderson, J. D. & Spencer, J. (2009). Gut-associated lymphoid tissue contains the molecular machinery to support T-cell-dependent and T-cell-independent class switch recombination. Mucosal Immunol 2, 495–503. - PubMed

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