82 research outputs found
HIV subject MDC are impaired in naïve CD4 T cell activation activity.
<p>Freshly prepared MDC from healthy control (n = 17) and HIV+ (n = 18) subjects were used in titrated numbers (<i>x</i>-axis) to activate one healthy control subject's allogeneic naive CD4 T-cells to produce IFN-γ or IL-2 (<i>y</i>-axis) in a 72-h culture performed in the absence (Medium, <b>panels </b><b><i>A</i></b><b>. and </b><b><i>C</i></b><b>.</b>), or presence of poly I∶C (<b>panels </b><b><i>B</i></b><b>. and </b><b><i>D</i></b><b>.</b>). Control cultures of TLR ligand and naïve CD4 T cells resulted in <3 sfu IFN-γ, and data shown are sfu above this background.</p
Increased PDL-1 and PDL-2 expression on HIV subject MDC.
<p>Isolated MDC were evaluated for expression of inhibitory molecules PDL-1 (<b>panel </b><b><i>A</i></b>) and PDL-2 (<b>panel </b><b><i>B</i></b>) in control (n = 10) and HIV+ (n = 10) subjects. Black lines represent median MFI value for each group. <b>Panel </b><b><i>C</i></b>. Association between PDL-1 MFI on MDC in HIV+ subjects and plasma HIV level. <b>Panel </b><b><i>D</i></b>. The percentage of Annexin V+ T-cells are shown following naive T-cell co-cultures with control vs. HIV+ subject MDC (10,000cells/well) in the absence and presence of poly I∶C.</p
HIV subject DC exhibit increased maturation.
<p><b>Panel </b><b><i>A</i></b>. Representative flow cytometric analysis of freshly isolated MDC and PDC from one healthy control. Mean Fluorescent Intensity (<b>MFI</b>) for HLA-DR, CD86, and CD83 were analyzed. <b>Panel </b><b><i>B</i></b>. Baseline activation/maturation phenotype of freshly isolated MDC and PDC from healthy control (n = 22) and HIV+ subjects (n = 24). The black line represents the median MFI value for each group for HLA-DR, CD86, and CD83.</p
Hydrogen peroxide impairs IL-7 induction of P-STAT5 in T cell subsets.
<p>PBMC from healthy control donors were pre-incubated with H<sub>2</sub>O<sub>2</sub> at the indicated concentrations for 30 minutes prior to stimulation with IL-7 (1 ng/ml). The sequential gating strategy included gating on live lymphocytes by forward and side scatter properties, removal of doublets (FSC-A vs FSC-H), gating on CD3+CD4+ cells or CD3+CD8+ cells and then gating on cells based on CD45RA and CD27 co-expression patterns to define subsets. Medians and inter-quartile ranges of P-STAT5+ cells within indicated subsets are represented by box-and-whisker plots for CD4+ T cells (n = 10) and CD8+ T cells (n = 6). Differences between H<sub>2</sub>O<sub>2</sub>-treated cells and cells incubated in medium alone were statistically significant at each concentration and for each cell type (p values <0.05). (A) P-STAT5 expression is shown in representative histograms of CD4+ (left column) and CD8+ (right column) T cells from either PBMC cell cultures (top) or purified T cells (>98% purity; bottom). Cells were incubated with H<sub>2</sub>O<sub>2</sub> at the indicated molar concentrations for 30 min. or incubated in medium alone. Cells were then stimulated with IL-7 and assessed for intracellular P-STAT5 expression (1 ng/ml). Dashed histograms represent P-STAT5 expression in unstimulated cells. Data are representative of experiments from 3 different donors (B). PBMC from 5 different donors were used to assess the effects of H<sub>2</sub>O<sub>2</sub> on T cell apoptosis and P-STAT5 induction. The percentages of total apoptotic CD4+ and CD8+ T cells were determined by annexin V staining during culture of PBMC with 98 µM H<sub>2</sub>O<sub>2.</sub> These percentages are compared to the percent inhibition of P-STAT5 signaling in PBMC that was observed after stimulation with IL-7 in the presence of 98 µM H<sub>2</sub>O<sub>2</sub> (C). Note that the proportions of cells that show evidence of apoptosis are far lower than the proportions of cells with impaired P-STAT5 induction.</p
Spearman correlations for relationships between MDA serum concentrations and measures of CD127 expression in T cell subsets.
<p>Spearman correlations for relationships between MDA serum concentrations and measures of CD127 expression in T cell subsets.</p
DC TLR ligand responsiveness.
<p><b>Panels </b><b><i>A and B</i></b>. CD86 MFI and HLA-DR MFI on isolated MDC following overnight culture in presence of Medium and poly I∶C stimulation in healthy control (n = 21) and HIV+ subjects (n = 18). The p value in panel B is for comparison between the delta HLA-DR MFI for controls and HIV infected subjects . IL-6 production from isolated MDC on a subset of the same subjects (n = 18 control and n = 16 HIV+) (<b>panel </b><b><i>C</i></b>) following overnight culture in absence and presence of poly I∶C stimulation. IL-6 production from isolated PDC on a subset of the same subjects (n = 17 control and n = 17 HIV+) (<b>panel </b><b><i>D</i></b>) following overnight culture in absence and presence of R848 stimulation. Healthy control (n = 17) and HIV+ subject (n = 16) PDC IFN-α production in response to overnight R848 stimulation is shown in <b>panel </b><b><i>E</i></b>. <b>Panel .</b> Association between MDC spontaneous IL-6 production and HIV plasma level.</p
P-STAT5 induction is related to CD127 expression in CD8+ T cells.
<p>The relationship between IL-7-induced P-STAT+ cells and the percentage of CD127+ T cells in whole blood are shown for CD4 cells (left column) and CD8 cells (right column) within the indicated subsets. Open symbols represent aviremic subjects and closed symbols represent viremic subjects. Correlation coefficients and P values were determined by Spearman's correlations.</p
Increased sCD14 and MDA adducts in serum of HIV+ donors.
<p>Serum samples from HIV+ and HIV- donors were stored at −80°C until analyzed in batch with commercial ELISA kits. Data are shown comparing cytokine levels in all HIV+ donors to controls (A) or comparing viremic, aviremic and controls (B). Statistical significance was determined by Mann Whitney tests (A) or by Kruskal-Wallis Test for multi-group analysis and Mann Whitney (B).</p
Serum concentrations of MDA adducts are inversely related to IL-7-induction of P-STAT5.
<p>Percent induction of P-STAT5 in the indicated T cell subsets was plotted against serum MDA adducts for each subject. Open symbols represent aviremic subjects and closed symbols represent viremic subjects. Correlation coefficients and P values were determined by Spearman's correlations.</p
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