Journal: Cell Communication and Signaling : CCS

Article Title: Subsequent malaria enhances virus-specific T cell immunity in SIV-infected Chinese rhesus macaques

doi: 10.1186/s12964-022-00910-7

Figure Lengend Snippet: T-cell repertoire diversity and SIV-specific immune response in malaria- and SIV-infected monkeys. A Diversity index differences with respect to the 8th week of SIV infection (corresponding to malaria introduction in the S + P group). Repertoire diversity was restored in the S + P group but not in the P + S group. A significant difference between the S + P and P + S groups was observed for the Shannon diversity index. P -values computed using ANOVA. B Ratio of SIV-specific clonotype frequency with respect to the control point (8th week of SIV infection, corresponding to malaria introduction in the S + P group). The S + P group, but not the P + S group, was characterized by a persistently higher SIV-specific clonotype frequency. P -values were calculated using ANOVA. C New SIV-specific clonotype production. Fractions of existing (detected at previous sampling point) and new SIV-specific clonotypes shown for each monkey in the P + S, S and S + P groups. A significant increase in new SIV-specific clonotype production was observed in the S + P group during the whole period of Plasmodium infection (at 11–17 weeks and 50 weeks post SIV infection). D Frequency of IFN-g-producing SIV Gag-specific PBMCs. Number of IFN-γ-producing PBMCs against pooled SIV Gag peptides analyzed by ELISPOT. The sample sizes were 6, 4 and 5 on day 49 and 4, 3, and 5 on day 161 of the S, P + S and S + P groups, respectively. SFC: spot-forming cells

Article Snippet: The plates were washed with PBST (PBS containing 0.05% Tween-20), and a biotinylated anti-IFN-γ polyclonal detector antibody (BD Biosciences) was added.

Techniques: Infection, Sampling, Enzyme-linked Immunospot

Journal: Frontiers in Immunology

Article Title: Immune Protection of SIV Challenge by PD-1 Blockade During Vaccination in Rhesus Monkeys

doi: 10.3389/fimmu.2018.02415

Figure Lengend Snippet: Modulating SIV-specific T cell immune responses by PD-1 blockade in vitro . SIV-specific T cells from chronic SIV-infected monkeys were stimulated with either SIV peptide pool alone or with SIV peptides in combination with genolimzumab, and then detected by IFN-γ-mediated enzyme-linked immunospot (ELISPOT) assay (A) . The above samples were also analyzed for IFN-γ expression (B) and IL-2 (C) expression by Q-PCR. Moreover, the frequency of SIV Gag-specific cytokine-secreting CD3+ T cells was further detected by intracellular cytokine staining. IFN-γ+ CD3+ T cells (D) , IL-2+CD3 T cells (E) , and TNF-α+CD3+ T cells (F) are indicated, respectively. The final data are represented as the mean ± SEM from at least triplicate experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: SIV peptide pools were added into cells for 20–24 h for stimulation, and then a polyclonal anti-monkey IFN-γ biotinylated detector antibody (BD Pharmingen) was added.

Techniques: In Vitro, Infection, Enzyme-linked Immunospot, Expressing, Staining

Journal: Frontiers in Immunology

Article Title: Immune Protection of SIV Challenge by PD-1 Blockade During Vaccination in Rhesus Monkeys

doi: 10.3389/fimmu.2018.02415

Figure Lengend Snippet: SIV-specific T cell immune responses regulated by PD-1 blockade in vivo in rhesus monkeys. Rhesus monkeys were intravenously injected with anti-PD-1 antibody during the SIV vaccine immunization schedule as described in Figure . For assessing total cellular immune responses induced by SIV vaccine regimen, IFN-γ-secreting cells were detected by ELISPOT assay over the course of immunization. (A) Representative data to illustrate IFN-γ-mediated ELISPOT cellular immune responses against SIV structure proteins (Gag, Pol, and Env), for each monkey at 9 weeks after first immunization. (B) Dynamic analysis of SIV-specific IFN-γ-mediated ELISPOT cellular immune responses over time until 42 weeks after first immunization. Data represent spot-forming cells (SFC) per million PBMCs. For assessing the polyfunctionality of SIV-specific CD8+ T cells, IFN-γ-and TNF-α-secreting cells were detected by intracellular cytokine staining (ICS) assays. (C) Representative flow cytometry dot plots for IFN-γ-and TNF-α-secreting CD8+ T cell populations. (D) Statistical analysis for the frequency of IFN-γ-and TNF-α-secreting CD8+ T cells by ICS. CFSE staining was used for assessing the ability of SIV antigen-stimulated T cell proliferation, whereby PBMCs were incubated with CFSE stain and stimulated with SIV peptides. (E) Representative dot plots for specific CD8+ T cell proliferation by CFSE staining. Statistical analysis for the proliferation index of CD8+ T cells in response to SIV antigen stimulation (F) or PMA+I simulation (G) . The daughter cells from proliferation had lower fluorescence intensity (CFSE-low cells). DMSO treatment was used as a negative control. * P < 0.05; *** P < 0.001.

Article Snippet: SIV peptide pools were added into cells for 20–24 h for stimulation, and then a polyclonal anti-monkey IFN-γ biotinylated detector antibody (BD Pharmingen) was added.

Techniques: In Vivo, Injection, Enzyme-linked Immunospot, Staining, Flow Cytometry, Incubation, Fluorescence, Negative Control