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Long-term programming of antigen-specific immunity from gene expression signatures in the PBMC of rhesus macaques immunized with an SIV DNA vaccine.

Belisle SE, Yin J, Shedlock DJ, Dai A, Yan J, Hirao L, Kutzler MA, Lewis MG, Andersen H, Lank SM, Karl JA, O'Connor DH, Khan A, Sardesai N, Chang J, Aicher L, Palermo RE, Weiner DB, Katze MG, Boyer J - PLoS ONE (2011)

Bottom Line: We observed that the RANTES-adjuvanted animals were significantly better at suppressing viral replication during chronic infection and exhibited a distinct pattern of gene expression which included immune cell-trafficking and cell cycle genes.Furthermore, a greater percentage of vaccine-induced central memory CD8+ T-cells capable of an activated phenotype were detected in these animals as measured by activation analysis.Thus, co-immunization with the RANTES molecular adjuvant followed by EP led to the generation of cellular immunity that was transcriptionally distinct and had a greater protective efficacy than its DNA alone counterpart.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Washington, Seattle, Washington, United States of America.

ABSTRACT
While HIV-1-specific cellular immunity is thought to be critical for the suppression of viral replication, the correlates of protection have not yet been determined. Rhesus macaques (RM) are an important animal model for the study and development of vaccines against HIV/AIDS. Our laboratory has helped to develop and study DNA-based vaccines in which recent technological advances, including genetic optimization and in vivo electroporation (EP), have helped to dramatically boost their immunogenicity. In this study, RMs were immunized with a DNA vaccine including individual plasmids encoding SIV gag, env, and pol alone, or in combination with a molecular adjuvant, plasmid DNA expressing the chemokine ligand 5 (RANTES), followed by EP. Along with standard immunological assays, flow-based activation analysis without ex vivo restimulation and high-throughput gene expression analysis was performed. Strong cellular immunity was induced by vaccination which was supported by all assays including PBMC microarray analysis that identified the up-regulation of 563 gene sequences including those involved in interferon signaling. Furthermore, 699 gene sequences were differentially regulated in these groups at peak viremia following SIVmac251 challenge. We observed that the RANTES-adjuvanted animals were significantly better at suppressing viral replication during chronic infection and exhibited a distinct pattern of gene expression which included immune cell-trafficking and cell cycle genes. Furthermore, a greater percentage of vaccine-induced central memory CD8+ T-cells capable of an activated phenotype were detected in these animals as measured by activation analysis. Thus, co-immunization with the RANTES molecular adjuvant followed by EP led to the generation of cellular immunity that was transcriptionally distinct and had a greater protective efficacy than its DNA alone counterpart. Furthermore, activation analysis and high-throughput gene expression data may provide better insight into mechanisms of viral control than may be observed using standard immunological assays.

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Proliferative capacity of SIV-specific T-cells in vaccinated animals.PBMCs from DNA-vaccinated RMs were isolated 4 weeks following the final immunization and analyzed for proliferative capacity. (A) Proliferation by CD4+ (top) and CD8+ (bottom) T-cells from one representative animal in the DNA-vaccinated group that were incubated for 5 days in the presence of medium alone, SIV peptides (gag, env, or pol), or ConA. Pseudo-color dot plots are gated on live CD3+ cells and T-cell gates are displayed with numbers representing total percent of CFSEdim T-cells. (B) T-cell data for each peptide (medium alone values subtracted) is enumerated per immunization group and shown for CD4+ and CD8+ T-cells. Error bars represent SD; *P<0.05 per the unpaired t test.
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pone-0019681-g002: Proliferative capacity of SIV-specific T-cells in vaccinated animals.PBMCs from DNA-vaccinated RMs were isolated 4 weeks following the final immunization and analyzed for proliferative capacity. (A) Proliferation by CD4+ (top) and CD8+ (bottom) T-cells from one representative animal in the DNA-vaccinated group that were incubated for 5 days in the presence of medium alone, SIV peptides (gag, env, or pol), or ConA. Pseudo-color dot plots are gated on live CD3+ cells and T-cell gates are displayed with numbers representing total percent of CFSEdim T-cells. (B) T-cell data for each peptide (medium alone values subtracted) is enumerated per immunization group and shown for CD4+ and CD8+ T-cells. Error bars represent SD; *P<0.05 per the unpaired t test.

Mentions: We next investigated the ability of vaccine-induced SIV-specific T-cells to proliferate upon antigen stimulation. PBMCs were isolated from RMs four weeks after the fourth immunization, labeled with CFSE, and then stimulated for 5 days with growth medium alone, SIV peptides, or ConA. T-cell proliferation was measured by flow cytometry, which showed that both CD4+ and CD8+ T-cell proliferative responses were higher in the DNA alone group compared to those from the DNA+RANTES group (Fig. 2). In addition, higher average proliferative responses were observed in the CD8+ compartment and that similar to ELISpot data, the env- and pol-specific responses were more extensive than the gag-specific response. These data show that DNA vaccination generates SIV-specific immunity that is capable of proliferation upon restimulation and that these responses were larger in animal receiving the DNA vaccine alone than in those that were adjuvanted. Altogether, these data demonstrate differences in the magnitudes of SIV-specific immunity generated by immunization with the DNA vaccine alone or in combination with the RANTES molecular adjuvant as measured by standard immunological assays.


Long-term programming of antigen-specific immunity from gene expression signatures in the PBMC of rhesus macaques immunized with an SIV DNA vaccine.

Belisle SE, Yin J, Shedlock DJ, Dai A, Yan J, Hirao L, Kutzler MA, Lewis MG, Andersen H, Lank SM, Karl JA, O'Connor DH, Khan A, Sardesai N, Chang J, Aicher L, Palermo RE, Weiner DB, Katze MG, Boyer J - PLoS ONE (2011)

Proliferative capacity of SIV-specific T-cells in vaccinated animals.PBMCs from DNA-vaccinated RMs were isolated 4 weeks following the final immunization and analyzed for proliferative capacity. (A) Proliferation by CD4+ (top) and CD8+ (bottom) T-cells from one representative animal in the DNA-vaccinated group that were incubated for 5 days in the presence of medium alone, SIV peptides (gag, env, or pol), or ConA. Pseudo-color dot plots are gated on live CD3+ cells and T-cell gates are displayed with numbers representing total percent of CFSEdim T-cells. (B) T-cell data for each peptide (medium alone values subtracted) is enumerated per immunization group and shown for CD4+ and CD8+ T-cells. Error bars represent SD; *P<0.05 per the unpaired t test.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3119060&req=5

pone-0019681-g002: Proliferative capacity of SIV-specific T-cells in vaccinated animals.PBMCs from DNA-vaccinated RMs were isolated 4 weeks following the final immunization and analyzed for proliferative capacity. (A) Proliferation by CD4+ (top) and CD8+ (bottom) T-cells from one representative animal in the DNA-vaccinated group that were incubated for 5 days in the presence of medium alone, SIV peptides (gag, env, or pol), or ConA. Pseudo-color dot plots are gated on live CD3+ cells and T-cell gates are displayed with numbers representing total percent of CFSEdim T-cells. (B) T-cell data for each peptide (medium alone values subtracted) is enumerated per immunization group and shown for CD4+ and CD8+ T-cells. Error bars represent SD; *P<0.05 per the unpaired t test.
Mentions: We next investigated the ability of vaccine-induced SIV-specific T-cells to proliferate upon antigen stimulation. PBMCs were isolated from RMs four weeks after the fourth immunization, labeled with CFSE, and then stimulated for 5 days with growth medium alone, SIV peptides, or ConA. T-cell proliferation was measured by flow cytometry, which showed that both CD4+ and CD8+ T-cell proliferative responses were higher in the DNA alone group compared to those from the DNA+RANTES group (Fig. 2). In addition, higher average proliferative responses were observed in the CD8+ compartment and that similar to ELISpot data, the env- and pol-specific responses were more extensive than the gag-specific response. These data show that DNA vaccination generates SIV-specific immunity that is capable of proliferation upon restimulation and that these responses were larger in animal receiving the DNA vaccine alone than in those that were adjuvanted. Altogether, these data demonstrate differences in the magnitudes of SIV-specific immunity generated by immunization with the DNA vaccine alone or in combination with the RANTES molecular adjuvant as measured by standard immunological assays.

Bottom Line: We observed that the RANTES-adjuvanted animals were significantly better at suppressing viral replication during chronic infection and exhibited a distinct pattern of gene expression which included immune cell-trafficking and cell cycle genes.Furthermore, a greater percentage of vaccine-induced central memory CD8+ T-cells capable of an activated phenotype were detected in these animals as measured by activation analysis.Thus, co-immunization with the RANTES molecular adjuvant followed by EP led to the generation of cellular immunity that was transcriptionally distinct and had a greater protective efficacy than its DNA alone counterpart.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Washington, Seattle, Washington, United States of America.

ABSTRACT
While HIV-1-specific cellular immunity is thought to be critical for the suppression of viral replication, the correlates of protection have not yet been determined. Rhesus macaques (RM) are an important animal model for the study and development of vaccines against HIV/AIDS. Our laboratory has helped to develop and study DNA-based vaccines in which recent technological advances, including genetic optimization and in vivo electroporation (EP), have helped to dramatically boost their immunogenicity. In this study, RMs were immunized with a DNA vaccine including individual plasmids encoding SIV gag, env, and pol alone, or in combination with a molecular adjuvant, plasmid DNA expressing the chemokine ligand 5 (RANTES), followed by EP. Along with standard immunological assays, flow-based activation analysis without ex vivo restimulation and high-throughput gene expression analysis was performed. Strong cellular immunity was induced by vaccination which was supported by all assays including PBMC microarray analysis that identified the up-regulation of 563 gene sequences including those involved in interferon signaling. Furthermore, 699 gene sequences were differentially regulated in these groups at peak viremia following SIVmac251 challenge. We observed that the RANTES-adjuvanted animals were significantly better at suppressing viral replication during chronic infection and exhibited a distinct pattern of gene expression which included immune cell-trafficking and cell cycle genes. Furthermore, a greater percentage of vaccine-induced central memory CD8+ T-cells capable of an activated phenotype were detected in these animals as measured by activation analysis. Thus, co-immunization with the RANTES molecular adjuvant followed by EP led to the generation of cellular immunity that was transcriptionally distinct and had a greater protective efficacy than its DNA alone counterpart. Furthermore, activation analysis and high-throughput gene expression data may provide better insight into mechanisms of viral control than may be observed using standard immunological assays.

Show MeSH
Related in: MedlinePlus