Immunogenicity of inactivated rotavirus in rhesus monkey, and assessment of immunologic mechanisms

Figures

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  Figure 1.. Schedule of IRV immunization and blood collection. Monkeys were immunized with different dose (160 EU,320 EU and 640 EU) of IRV at weeks 0,4 or 0,4, and 8. Serum was collected from the two-dose immunization group was collected at weeks 4 and 6 for the determination of neutralizing antibodies and IgG and IgA antibodies. Serum was collected from the three-dose immunization group at weeks 4, 6 and 10 for the determination of neutralizing antibodies and IgG and IgA antibodies. Serum was collected at weeks 2, 6, 8, 12, 16, 24, and 28 in the two-dose immunization group and at weeks 2, 6, 10, 12, 28, and 52 for in the three-dose immunization group for immune persistence evaluation. The peripheral blood of monkeys 2 weeks after the whole immunization was collected to separate peripheral blood lymphocytes for IFN-γ ELISpot detection. The peripheral blood of the monkeys was collected for cytokine detection. PBMC samples from the peripheral blood of monkeys 5 days after each dose of immunization were collected for gene profile analysis. Red tubes and red hearts represent all blood collection time points. Specifically, red hearts represent gene expression profile detection time points,…

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  Figure 2.. Neutralizing antibody responses after vaccine immunization. (a) Neutralizing antibodies (GMT) of monkeys at week 4 and 6 of 160 EU, 320 EU, and 640 EU in the two-dose immunization group (n = 3). (b) Neutralizing antibodies (GMT) of monkeys at week 4, 6, and 10 of 160 EU, 320 EU, and 640 EU in the three-dose immunization group (n = 3); (c) Anti-rotavirus IgG antibody responses in the three-dose immunization group (n = 3); (d) Anti-rotavirus IgA antibody responses in the three-dose immunization group (n = 3). Error bars represent the standard error of each group.

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  Figure 3.. Neutralizing antibody responses in immune persistence evaluation. (a) Neutralizing antibodies (GMT) of monkeys in the two-dose immunization group at week 0, 2, 6, 8, 12, 16, 24, and 28 (n = 3). (b) Neutralizing antibodies (GMT) of monkeys in the three-dose immunization group at weeks 2, 6, 10,12, 28, and 52 (n = 3).

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  Figure 4.. Cellular immunoassay of monkeys after IRV vaccination conducted via IFN-γ ELISPOT detection and cytokine array. (a) VP7-specific IFN-γ response of monkeys on week 2 after the last vaccination. IFN-γ ELISPOT detection was performed according to the instructions of the Monkey IFN-γ ELISPOT PLUS(ALP) kit (MABTECH, 3421 M-4HST-2); (b) Cytokines (TNF-α, IL-6, IL-5, IL-4, IL-1β, IL-16, IL-15, IL-12p70, IFN-γ, and GM-CSF) of plasma were quantified via Quantibody® array (*: p < .05; **: p < .01; ***: p < .001);(c) Fold change of cytokines (TNF-α, IL-6, IL-5, IL-4, IL-1β, IL-16, IL-15, IL-12p70, IFN-γ, and GM-CSF) after IRV vaccination. (*: p < .05).

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  Figure 5.. Gene Ontology (GO) analysis of differentially expressed genes between pre-immunization and 5 days after the first immunization in the three-dose immunization group. Differential genes with a fold change > = 2.0 were chosen for GO analysis, including biological process, cellular component, and molecular function analysis.

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  Figure 6.. KEGG analysis of differentially expressed genes between pre-immunization and 5 days after the first immunization in the three-dose immunization group. Differential genes with a fold change > = 2.0 were chosen for KEGG analysis.

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  Figure 7.. Gene network analysis. The Chemokine-mediated signaling pathway and immune response pathway were mainly activated after IRV immunization. To identify the interaction of genes in these pathways, differentially genes (FC ≥ 2) in the chemokine-mediated signaling pathway and immune response term were chosen for network analysis. GeneMANIA was used to build a gene network according to the relationships among the genes. The interaction relationships of the differentially expressed genes will be represented as the gene network graphs. Each node in the graph represents one protein. The line between the nodes indicates the interaction between the two proteins. The protein with multiple connecting lines indicates the presence of multiple interactions, and the protein is the core protein. (a) Network analysis of differentially expressed genes (FC ≥ 2) in the ‘chemokine-mediated signaling ’pathway; (b) Network analysis of differentially expressed genes (FC ≥ 2) in the ‘immune response.