Journal: Nature Communications

Article Title: CTL-mediated immunotherapy can suppress SHIV rebound in ART-free macaques

doi: 10.1038/s41467-019-09725-6

Figure Lengend Snippet: Phenotypic and functional characteristics pre- and post CTL infusion. a Functional profiles of CD8 + T-cells pre- and post- CTL were evaluated by peptide antigen-specific responses and shown as the frequencies of Env-specific CD8 + T-cells producing CD107a, IFN-γ, MIP-1β and TNF-α ( p < 0.0001 using Wilcoxon signed-rank test, n = 6). b Activation profile of Env-specific, IFN-γ-producing CD8 + T-cells for both pre- and post-infusion responses, based on surface expression of CD69, HLADR, CCR5 and intracellular expression of Ki67 (* p < 0.05 and ** p < 0.01 using unpaired Students t test, Mean ± s.d. are presented in all graphs, n = 6). c Env-peptide functional profiles of CD8 + T-cells in controllers, non-controllers and untreated animals ( p < 0.0001 using Wilcoxon signed-rank test, n = 8). d Env-specific, IFN-γ + CD3 + CD8 + central memory T cells pre-and post-CTL infusion among controllers, non-controllers and untreated animals ( p = 0.038 using paired Students t-test, n = 8). e PD-1 + Ki67 + expression on Env-specific CD8 + T-cells pre- and post-CTL infusion among controllers, non-controllers and untreated animals ( p = 0.01 using paired Students t-test, n = 8). f Env-specific CD8 + T-cells expressing PD-1 + Ki67 + were positively correlated with viral load post infusion (Spearman’s correlation coefficient (R) and P values are indicated. Note that x-axis is log 10 scales, n = 8). g The SHIV-suppressive activity of circulating CD8 + T-cells from controllers and non-controllers were measured on autologous activated CD4 + T-cells infected with SHIV-162P3. SHIV suppression was reported as the Log inhibition in p27 titers in CD4 + T-cells supernatants when autologous ex vivo Env-stimulated CD8 + T-cells were added at a 1:1 ratio to the culture ( p = 0.0001 using unpaired Students t test, Mean ± s.d. is presented, n = 6). h Relationship between SHIV-suppressive activity (% inhibition) of circulating CD8 + T-cells and set point of viral load (Spearman’s correlation coefficient (R) and P values are indicated. Note that x- and y-axis are log 10 scales, n = 6)

Article Snippet: Following two rounds of peptide stimulation, the Env-specific CD8 + T cells were isolated by non-human primate CD8 + T-cell isolation kit (Miltenyi Biotec, Catalogue# 130-092-143) and expanded using the Rapid Expansion Protocol in a sterile 25-cm 2 tissue culture flasks.

Techniques: Functional Assay, Activation Assay, Expressing, Activity Assay, Infection, Inhibition, Ex Vivo

Journal: Nature Communications

Article Title: CTL-mediated immunotherapy can suppress SHIV rebound in ART-free macaques

doi: 10.1038/s41467-019-09725-6

Figure Lengend Snippet: Phenotypic and functional characterization of CTL-lines. a In vitro specific antigen recognition of CTL-lines. Data is shown as Grand median for each group. Each symbol represented one animal ( n = 6). b P27 titers in the culture supernatants of PHA-activated CD4 + T-cells infected in vitro with replicative SHIV-SF162P3, in the absence (CD4 alone) or presence of autologous CTL Lines (1:1, 2:1 and 5:1 of ET ratio). Each symbol represented one animal ( n = 6). c The capacity of CTL-lines to suppress SHIV replication in autologous CD4 + T-cells. The Log of p27 decreased when CD4 + T-cells were cocultured with CTL-lines. Each symbol represented one animal ( n = 6). d Correlation between cytotoxic capacity (% inhibition) and frequencies of Env-specific IFN- γ producing (Spearman’s correlation coefficient (R) and P values are indicated. Note that y-axis is log 10 scales, n = 6). e Memory phenotypic characterization of CTL-lines. Data is shown as Grand median for each group. Each symbol represented one animal ( n = 6). f Relationship between PBMCs Env-specific, IFN-γ + CD3 + CD8 + central memory T cells post-infusion and Env-specific memory response of CTL-lines (Spearman’s correlation coefficient (R) and P values are indicated, n = 6)

Article Snippet: Following two rounds of peptide stimulation, the Env-specific CD8 + T cells were isolated by non-human primate CD8 + T-cell isolation kit (Miltenyi Biotec, Catalogue# 130-092-143) and expanded using the Rapid Expansion Protocol in a sterile 25-cm 2 tissue culture flasks.

Techniques: Functional Assay, In Vitro, Infection, Inhibition

Journal: Nature Communications

Article Title: CTL-mediated immunotherapy can suppress SHIV rebound in ART-free macaques

doi: 10.1038/s41467-019-09725-6

Figure Lengend Snippet: Kinetics of Env-specific immune response and the correlates of virologic control. a The frequency of circulating antigen-specific CD8 + T-cells in PBMCs was measured at the indicated time points from SHIV controllers (Animals 1–3, n = 3), non-controllers (Animals 4–6, n = 3) and untreated animals (Animals 7 and 8, n = 2). y -axis, from Lift to Right, frequency of HIV CN54 Env-specific CD8 + T-cell producing IFN-γ + and frequency of SIV mac239 Gag-specific CD8 + T-cell producing IFN-γ + . wk, weeks. b Correlations between Env-specific CD8 + T-cell responses at 2 weeks after CTL infusion and set point log SHIV RNA after CTL infusion (Spearman’s correlation coefficient (R) and P values are indicated. Note that x-axis is log 10 scales, n = 8). c Depletion of CD8 + T cells in controller macaques results in the rapid induction of plasma viremia. CD8 + T-cell count and plasma viral load were shown in SHIV controller macaques following CD8 depletion (Animal 1 and 3, n = 2). x-axis, days post- ART interruption. The shaded gray indicated time points post-CD8 depletion

Article Snippet: Following two rounds of peptide stimulation, the Env-specific CD8 + T cells were isolated by non-human primate CD8 + T-cell isolation kit (Miltenyi Biotec, Catalogue# 130-092-143) and expanded using the Rapid Expansion Protocol in a sterile 25-cm 2 tissue culture flasks.

Techniques: Control, Clinical Proteomics, Cell Counting