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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 is associated with differential regulation of cell lysis pathways at 8 months post-vaccination.(A) IPA networks indicate the relationship between differentially expressed genes related to cytolysis from analysis of the genes that significantly distinguished the treatment groups only at peak viremia by ANOVA. Molecule shading reflects the average logratio of expression for each group. Red represents an average increase in expression, green represents a decrease in average expression. (B) Heatmap indicating the expression of cytolysis genes at 8 months post-vaccination. Genes shown in red have higher expression than their genetically matched mock, gene shown in green have lower expression than their genetically matched mock. Clustering was by Hierarchial algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.4 logratio to mock.
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pone-0019681-g008: Vaccination is associated with differential regulation of cell lysis pathways at 8 months post-vaccination.(A) IPA networks indicate the relationship between differentially expressed genes related to cytolysis from analysis of the genes that significantly distinguished the treatment groups only at peak viremia by ANOVA. Molecule shading reflects the average logratio of expression for each group. Red represents an average increase in expression, green represents a decrease in average expression. (B) Heatmap indicating the expression of cytolysis genes at 8 months post-vaccination. Genes shown in red have higher expression than their genetically matched mock, gene shown in green have lower expression than their genetically matched mock. Clustering was by Hierarchial algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.4 logratio to mock.

Mentions: Next, we examined the biological functions of the genes that distinguished the treatment groups only at 8 months post-vaccination (Table S2). Differences in the expression of these genes suggest a long-term transcriptional reprogramming of cells within the PBMC following vaccination that were not maintained at peak viremia. We found that vaccination was associated with an up-regulation of cytolysis-related gene expression following POL stimulation (Figure 8). A qualitative examination of the expression of these genes at 10 days post-challenge suggested that these genes did not pass the statistical thresholds at peak viremia because the expression of these genes was generally up-regulated in all three groups at peak viremia (Data not shown).


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 is associated with differential regulation of cell lysis pathways at 8 months post-vaccination.(A) IPA networks indicate the relationship between differentially expressed genes related to cytolysis from analysis of the genes that significantly distinguished the treatment groups only at peak viremia by ANOVA. Molecule shading reflects the average logratio of expression for each group. Red represents an average increase in expression, green represents a decrease in average expression. (B) Heatmap indicating the expression of cytolysis genes at 8 months post-vaccination. Genes shown in red have higher expression than their genetically matched mock, gene shown in green have lower expression than their genetically matched mock. Clustering was by Hierarchial algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.4 logratio to mock.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0019681-g008: Vaccination is associated with differential regulation of cell lysis pathways at 8 months post-vaccination.(A) IPA networks indicate the relationship between differentially expressed genes related to cytolysis from analysis of the genes that significantly distinguished the treatment groups only at peak viremia by ANOVA. Molecule shading reflects the average logratio of expression for each group. Red represents an average increase in expression, green represents a decrease in average expression. (B) Heatmap indicating the expression of cytolysis genes at 8 months post-vaccination. Genes shown in red have higher expression than their genetically matched mock, gene shown in green have lower expression than their genetically matched mock. Clustering was by Hierarchial algorithm with average link criteria and cosine similarity measure. Color saturation was set at +/−0.4 logratio to mock.
Mentions: Next, we examined the biological functions of the genes that distinguished the treatment groups only at 8 months post-vaccination (Table S2). Differences in the expression of these genes suggest a long-term transcriptional reprogramming of cells within the PBMC following vaccination that were not maintained at peak viremia. We found that vaccination was associated with an up-regulation of cytolysis-related gene expression following POL stimulation (Figure 8). A qualitative examination of the expression of these genes at 10 days post-challenge suggested that these genes did not pass the statistical thresholds at peak viremia because the expression of these genes was generally up-regulated in all three groups at peak viremia (Data not shown).

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