Limits...
Prime-boost vaccination using chemokine-fused gp120 DNA and HIV envelope peptides activates both immediate and long-term memory cellular responses in rhesus macaques.

Qin H, Nehete PN, He H, Nehete B, Buchl S, Cha SC, Sastry JK, Kwak LW - J. Biomed. Biotechnol. (2010)

Bottom Line: HIV vaccine candidates with improved immunogenicity and induction of mucosal T-cell immunity are needed.With a peptide-cocktail vaccine containing a set of conserved polypeptides of HIV-1 envelope protein, given by nasogastric administration, primed T-cell immunity was significantly boosted.Therefore, data from this investigation offer proof-of-principle for potential effectiveness of the prime-boost strategy with a chemokine-fused gp120 DNA and warrant further testing in the nonhuman primate models for developing as a potential HIV vaccine candidate in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Lymphoma and Myeloma, M. D. Anderson Cancer Center, The University of Texas, Houston, TX 77030, USA.

ABSTRACT
HIV vaccine candidates with improved immunogenicity and induction of mucosal T-cell immunity are needed. A prime-boost strategy using a novel HIV glycoprotein 120 DNA vaccine was employed to immunize rhesus macaques. The DNA vaccine encoded a chimeric gp120 protein in fusion with monocyte chemoattractant protein-3, which was hypothesized to improve the ability of antigen-presenting cells to capture viral antigen through chemokine receptor-mediated endocytosis. DNA vaccination induced virus-reactive T cells in peripheral blood, detectable by T cell proliferation, INFgamma ELISPOT and sustained IL-6 production, without humoral responses. With a peptide-cocktail vaccine containing a set of conserved polypeptides of HIV-1 envelope protein, given by nasogastric administration, primed T-cell immunity was significantly boosted. Surprisingly, long-term and peptide-specific mucosal memory T-cell immunity was detected in both vaccinated macaques after one year. Therefore, data from this investigation offer proof-of-principle for potential effectiveness of the prime-boost strategy with a chemokine-fused gp120 DNA and warrant further testing in the nonhuman primate models for developing as a potential HIV vaccine candidate in humans.

Show MeSH

Related in: MedlinePlus

Immunization of rhesus macaques with plasmid DNA encoding MCP3-gp120 fusion primed viral-reactive T-cell immunity, which was further boosted by HIV-1 envelope peptide-cocktail vaccine. The DNA priming schedule included four rounds of immunization at 1-month intervals. Two rhesus macaques were immunized with 20 μg of the plasmid DNA by gene gun injection at weeks 0, 4, 8, and 12. The boosting schedule included a peptide-cocktail vaccine that contains six highly conserved peptides from the HIV-1 envelope protein. After a rest period of 20 weeks following the final round of DNA vaccination to allow for the establishment of memory T cells, the peptide-cocktail boosts were administered by intranasal route along with a mutant cholera toxin on Weeks 32 and 37.  Peripheral blood mononuclear cells collected after DNA vaccination revealed viral-reactive T-cell immunity as evidenced by the cell proliferation (a) and IFN-γ producing cells (b) in response to in vitro stimulation with cell-free, heat-inactivated SHIV89.6P antigen-specific T-cell immunity was boosted by the peptide-cocktail vaccine, especially in #J160 showing significant elevation of IFN-γ producing cells in response to heat-inactivated SHIV89.6P) (b). The immunogenicity of the peptide vaccine was confirmed by analysis of peptide-specific T-cell immunity (c). Vaccine-induced cytokine production was examined using LINCOplex multiple cytokine luminescent assay. Among the cytokines assayed, IL-6 was predominantly detected in both macaques after DNA vaccination at Week 12. Virus-reactive IL-6 secretion was more apparent in monkey #J160, particularly during the period of peptide-cocktail boost.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2864514&req=5

fig1: Immunization of rhesus macaques with plasmid DNA encoding MCP3-gp120 fusion primed viral-reactive T-cell immunity, which was further boosted by HIV-1 envelope peptide-cocktail vaccine. The DNA priming schedule included four rounds of immunization at 1-month intervals. Two rhesus macaques were immunized with 20 μg of the plasmid DNA by gene gun injection at weeks 0, 4, 8, and 12. The boosting schedule included a peptide-cocktail vaccine that contains six highly conserved peptides from the HIV-1 envelope protein. After a rest period of 20 weeks following the final round of DNA vaccination to allow for the establishment of memory T cells, the peptide-cocktail boosts were administered by intranasal route along with a mutant cholera toxin on Weeks 32 and 37. Peripheral blood mononuclear cells collected after DNA vaccination revealed viral-reactive T-cell immunity as evidenced by the cell proliferation (a) and IFN-γ producing cells (b) in response to in vitro stimulation with cell-free, heat-inactivated SHIV89.6P antigen-specific T-cell immunity was boosted by the peptide-cocktail vaccine, especially in #J160 showing significant elevation of IFN-γ producing cells in response to heat-inactivated SHIV89.6P) (b). The immunogenicity of the peptide vaccine was confirmed by analysis of peptide-specific T-cell immunity (c). Vaccine-induced cytokine production was examined using LINCOplex multiple cytokine luminescent assay. Among the cytokines assayed, IL-6 was predominantly detected in both macaques after DNA vaccination at Week 12. Virus-reactive IL-6 secretion was more apparent in monkey #J160, particularly during the period of peptide-cocktail boost.

Mentions: We used nonhuman primates to evaluate the translational potential for the chemokine-fusion strategy in the development of a novel HIV vaccine. Two macaques immunized with the plasmid DNA encoding MCP3-gp120 fusions demonstrated virus-reactive T-cell immunity at week 12, evidenced by the proliferation of T cells (Figure 1(a)) and IFN-γ producing cells in response to in vitro stimulation with heat-inactivated SHIV89.6P (Figure 1(b)). Peptide-cocktail vaccine boosted the viral-reactive T-cell immunity, as evidenced by the significant elevation of IFN-γ producing cells in response to the heat-inactivated SHIV89.6P in one of the two monkeys (Figure 1(b) #J160). The peptide-cocktail-specific response was readily detected after peptide boost, confirming the immunogenicity of the peptide-cocktail vaccine (Figure 1(c)). The macaque (#J8) that was weakly responsive to the peptide boost (Figure 1(b) #J8) also showed a low magnitude of peptide-cocktail-specific systemic cellular responses (Figure 1(c)). PBMC cultured with medium or treated with irrelevant human papilloma virus peptide failed to show positive responses (data not shown). We were unable to detect antibody responses, either by neutralization assay or by Western blot (data not shown).


Prime-boost vaccination using chemokine-fused gp120 DNA and HIV envelope peptides activates both immediate and long-term memory cellular responses in rhesus macaques.

Qin H, Nehete PN, He H, Nehete B, Buchl S, Cha SC, Sastry JK, Kwak LW - J. Biomed. Biotechnol. (2010)

Immunization of rhesus macaques with plasmid DNA encoding MCP3-gp120 fusion primed viral-reactive T-cell immunity, which was further boosted by HIV-1 envelope peptide-cocktail vaccine. The DNA priming schedule included four rounds of immunization at 1-month intervals. Two rhesus macaques were immunized with 20 μg of the plasmid DNA by gene gun injection at weeks 0, 4, 8, and 12. The boosting schedule included a peptide-cocktail vaccine that contains six highly conserved peptides from the HIV-1 envelope protein. After a rest period of 20 weeks following the final round of DNA vaccination to allow for the establishment of memory T cells, the peptide-cocktail boosts were administered by intranasal route along with a mutant cholera toxin on Weeks 32 and 37.  Peripheral blood mononuclear cells collected after DNA vaccination revealed viral-reactive T-cell immunity as evidenced by the cell proliferation (a) and IFN-γ producing cells (b) in response to in vitro stimulation with cell-free, heat-inactivated SHIV89.6P antigen-specific T-cell immunity was boosted by the peptide-cocktail vaccine, especially in #J160 showing significant elevation of IFN-γ producing cells in response to heat-inactivated SHIV89.6P) (b). The immunogenicity of the peptide vaccine was confirmed by analysis of peptide-specific T-cell immunity (c). Vaccine-induced cytokine production was examined using LINCOplex multiple cytokine luminescent assay. Among the cytokines assayed, IL-6 was predominantly detected in both macaques after DNA vaccination at Week 12. Virus-reactive IL-6 secretion was more apparent in monkey #J160, particularly during the period of peptide-cocktail boost.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2864514&req=5

fig1: Immunization of rhesus macaques with plasmid DNA encoding MCP3-gp120 fusion primed viral-reactive T-cell immunity, which was further boosted by HIV-1 envelope peptide-cocktail vaccine. The DNA priming schedule included four rounds of immunization at 1-month intervals. Two rhesus macaques were immunized with 20 μg of the plasmid DNA by gene gun injection at weeks 0, 4, 8, and 12. The boosting schedule included a peptide-cocktail vaccine that contains six highly conserved peptides from the HIV-1 envelope protein. After a rest period of 20 weeks following the final round of DNA vaccination to allow for the establishment of memory T cells, the peptide-cocktail boosts were administered by intranasal route along with a mutant cholera toxin on Weeks 32 and 37. Peripheral blood mononuclear cells collected after DNA vaccination revealed viral-reactive T-cell immunity as evidenced by the cell proliferation (a) and IFN-γ producing cells (b) in response to in vitro stimulation with cell-free, heat-inactivated SHIV89.6P antigen-specific T-cell immunity was boosted by the peptide-cocktail vaccine, especially in #J160 showing significant elevation of IFN-γ producing cells in response to heat-inactivated SHIV89.6P) (b). The immunogenicity of the peptide vaccine was confirmed by analysis of peptide-specific T-cell immunity (c). Vaccine-induced cytokine production was examined using LINCOplex multiple cytokine luminescent assay. Among the cytokines assayed, IL-6 was predominantly detected in both macaques after DNA vaccination at Week 12. Virus-reactive IL-6 secretion was more apparent in monkey #J160, particularly during the period of peptide-cocktail boost.
Mentions: We used nonhuman primates to evaluate the translational potential for the chemokine-fusion strategy in the development of a novel HIV vaccine. Two macaques immunized with the plasmid DNA encoding MCP3-gp120 fusions demonstrated virus-reactive T-cell immunity at week 12, evidenced by the proliferation of T cells (Figure 1(a)) and IFN-γ producing cells in response to in vitro stimulation with heat-inactivated SHIV89.6P (Figure 1(b)). Peptide-cocktail vaccine boosted the viral-reactive T-cell immunity, as evidenced by the significant elevation of IFN-γ producing cells in response to the heat-inactivated SHIV89.6P in one of the two monkeys (Figure 1(b) #J160). The peptide-cocktail-specific response was readily detected after peptide boost, confirming the immunogenicity of the peptide-cocktail vaccine (Figure 1(c)). The macaque (#J8) that was weakly responsive to the peptide boost (Figure 1(b) #J8) also showed a low magnitude of peptide-cocktail-specific systemic cellular responses (Figure 1(c)). PBMC cultured with medium or treated with irrelevant human papilloma virus peptide failed to show positive responses (data not shown). We were unable to detect antibody responses, either by neutralization assay or by Western blot (data not shown).

Bottom Line: HIV vaccine candidates with improved immunogenicity and induction of mucosal T-cell immunity are needed.With a peptide-cocktail vaccine containing a set of conserved polypeptides of HIV-1 envelope protein, given by nasogastric administration, primed T-cell immunity was significantly boosted.Therefore, data from this investigation offer proof-of-principle for potential effectiveness of the prime-boost strategy with a chemokine-fused gp120 DNA and warrant further testing in the nonhuman primate models for developing as a potential HIV vaccine candidate in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Lymphoma and Myeloma, M. D. Anderson Cancer Center, The University of Texas, Houston, TX 77030, USA.

ABSTRACT
HIV vaccine candidates with improved immunogenicity and induction of mucosal T-cell immunity are needed. A prime-boost strategy using a novel HIV glycoprotein 120 DNA vaccine was employed to immunize rhesus macaques. The DNA vaccine encoded a chimeric gp120 protein in fusion with monocyte chemoattractant protein-3, which was hypothesized to improve the ability of antigen-presenting cells to capture viral antigen through chemokine receptor-mediated endocytosis. DNA vaccination induced virus-reactive T cells in peripheral blood, detectable by T cell proliferation, INFgamma ELISPOT and sustained IL-6 production, without humoral responses. With a peptide-cocktail vaccine containing a set of conserved polypeptides of HIV-1 envelope protein, given by nasogastric administration, primed T-cell immunity was significantly boosted. Surprisingly, long-term and peptide-specific mucosal memory T-cell immunity was detected in both vaccinated macaques after one year. Therefore, data from this investigation offer proof-of-principle for potential effectiveness of the prime-boost strategy with a chemokine-fused gp120 DNA and warrant further testing in the nonhuman primate models for developing as a potential HIV vaccine candidate in humans.

Show MeSH
Related in: MedlinePlus