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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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Vaccination corresponded to differential gene expression response to SIV Pol stimulation.(A) 563 gene sequences were differentially expressed at 8 months post-vaccination by ANOVA while (B) 699 gene sequences were differentially expressed among the three treatment groups at 10 days post-challenge by ANOVA. Genes shown in red have higher expression than their genetically matched mock, while genes shown in green have lower expression. Clustering was by Hierarchical algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.6 logratio to mock.
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pone-0019681-g005: Vaccination corresponded to differential gene expression response to SIV Pol stimulation.(A) 563 gene sequences were differentially expressed at 8 months post-vaccination by ANOVA while (B) 699 gene sequences were differentially expressed among the three treatment groups at 10 days post-challenge by ANOVA. Genes shown in red have higher expression than their genetically matched mock, while genes shown in green have lower expression. Clustering was by Hierarchical algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.6 logratio to mock.

Mentions: To explore the long-term effect of vaccination on response to SIV antigen stimulation and possible mechanisms underlying differential response to SIV challenge, gene expression in response to SIV pol peptide stimulation in fresh PBMCs was measured using DNA microarray at eight months following the last vaccination (prior to SIV challenge) and at peak viremia (10 days following SIV challenge). We applied statistical methods to distinguish treatment groups (n = 5–6 animals per group) by gene expression. At eight months following vaccination, there were 563 gene sequences that distinguished the three groups and at peak viremia there were 699 gene sequences that distinguished the three groups (Figure 5). The majority of gene expression differences were between the control group and any of the vaccinated groups, with quantitatively fewer differences distinguishing the vaccine groups. These general gene expression signatures mirror the standard immunological assays, suggesting not only the long-term impact on antigen specific response to vaccination but also relative subtly in the differences between the vaccinated groups.


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)

Vaccination corresponded to differential gene expression response to SIV Pol stimulation.(A) 563 gene sequences were differentially expressed at 8 months post-vaccination by ANOVA while (B) 699 gene sequences were differentially expressed among the three treatment groups at 10 days post-challenge by ANOVA. Genes shown in red have higher expression than their genetically matched mock, while genes shown in green have lower expression. Clustering was by Hierarchical algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.6 logratio to mock.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0019681-g005: Vaccination corresponded to differential gene expression response to SIV Pol stimulation.(A) 563 gene sequences were differentially expressed at 8 months post-vaccination by ANOVA while (B) 699 gene sequences were differentially expressed among the three treatment groups at 10 days post-challenge by ANOVA. Genes shown in red have higher expression than their genetically matched mock, while genes shown in green have lower expression. Clustering was by Hierarchical algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.6 logratio to mock.
Mentions: To explore the long-term effect of vaccination on response to SIV antigen stimulation and possible mechanisms underlying differential response to SIV challenge, gene expression in response to SIV pol peptide stimulation in fresh PBMCs was measured using DNA microarray at eight months following the last vaccination (prior to SIV challenge) and at peak viremia (10 days following SIV challenge). We applied statistical methods to distinguish treatment groups (n = 5–6 animals per group) by gene expression. At eight months following vaccination, there were 563 gene sequences that distinguished the three groups and at peak viremia there were 699 gene sequences that distinguished the three groups (Figure 5). The majority of gene expression differences were between the control group and any of the vaccinated groups, with quantitatively fewer differences distinguishing the vaccine groups. These general gene expression signatures mirror the standard immunological assays, suggesting not only the long-term impact on antigen specific response to vaccination but also relative subtly in the differences between the vaccinated groups.

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