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<p>iNKT frequency was comparable among BD and DN groups (A). BD (n = 10) and DN (n = 10) showed similar CD161-expressing iNKT cell frequencies (B) and a significant increase in TNF production following PMA/ionomycin stimulation... more
<p>iNKT frequency was comparable among BD and DN groups (A). BD (n = 10) and DN (n = 10) showed similar CD161-expressing iNKT cell frequencies (B) and a significant increase in TNF production following PMA/ionomycin stimulation (p = .002 and p = .027 respectively). Despite a trend to higher spontaneous TNF release in BD patients (p = .075), comparable cytokine levels were recorded upon PMA/ionomycin (C). BD patients alone responded to PMA/ionomycin with significant IFN-γ production following stimulation (p = .0488) (D). Significantly higher TNF production was detected in BD subjects (p = .031) prior to α-GalCer stimulation. Upon α-GalCer stimulation, BD patients displayed a trend to significant increases in TNF release (p = .063), leading to higher cytokine levels in this population (p = .056) (E). No significant differences were noted in terms of IFN-γ production following α-GalCer, although BD patients tended to significant cytokine production (p = .063) (F). CD and DN showed comparable iNKT cell frequencies (G). CD (n = 10) and DN (n = 10) showed similar CD161-expressing iNKT cell frequencies (H). CD subjects showed higher TNF release both prior to (p = .005) and following stimulation with PMA/ionomycin (p = .029). Of note, DN patients alone responded to stimulation by significantly increasing TNF release from iNKT cells aspecific stimulation (p = .027) (I). In keeping with these results, the CD group displayed a trend to higher IFN-γ release after PMA/ionomycin stimulation (p = .052) (J). No statistical differences were noted between groups in terms of iNKT function following specific activation with α-GalCer (K, L). Horizontal lines indicate median values. Each symbol represents an individual.</p
<p>Gating strategy of flow cytometry analysis for staining of iNKT cell frequencies, phenotype and intracellular cytokine production in a representative HIV-positive individual (A); an example of staining for intracellular cytokines... more
<p>Gating strategy of flow cytometry analysis for staining of iNKT cell frequencies, phenotype and intracellular cytokine production in a representative HIV-positive individual (A); an example of staining for intracellular cytokines is also shown of a representative HIV-negative subject (B). PBMCs were gated on lymphocytes, and iNKT cells were visualized as CD3+, Vα24+ and CD1d-tetramer+. An example of CD161 surface staining is shown in the far right plot. iNKT frequency were comparable in DP and DN groups (C). iNKT cell phenotype was analyzed through the <i>ex vivo</i> expression of CD161 in DP (n = 10) and DN (n = 10) patients (D). DP subjects exhibited significantly higher levels of CD161 on iNKT cell surface compared to DN patients (p = .001). iNKT cell function was measured through the production of TNF and IFN-γ <i>ex vivo</i> (US) and following stimulation with PMA/ionomycin (n = 10 per group) (E, F) and α-GalCer (n = 5 per group) (G, H). Although DP and DN patients significantly increased TNF production upon PMA/ionomycin stimulation (p = .002 and p = .027 respectively), DP subjects showed higher TNF release both prior to (p = .049) and following PMA/ionomycin (E). Study groups exhibited similar frequencies of IFN-γ-producing iNKT cells both <i>ex vivo</i> and after stimulation with PMA/ionomycin (F). DP patients were characterized by significantly higher TNF release both prior to (p = .047) and following stimulation with α-GalCer (p = .021) (G). Similar results were obtained in terms of IFN-γ production, with a trend to higher cytokine production in DP subjects following iNKT-specific stimulation (p = .059) (H). FSC, Forward Scatter: SSC, Side Scatter. Each symbol represents an individual.</p
<p>A significant reduction of the CD4+ Tscm pool and no changes in the CD8+ Tscm subset was measured in the course of the study and both maintained lower frequencies compared to controls. In HIV-negative individuals, Tscm cells... more
<p>A significant reduction of the CD4+ Tscm pool and no changes in the CD8+ Tscm subset was measured in the course of the study and both maintained lower frequencies compared to controls. In HIV-negative individuals, Tscm cells correlated negatively with naïve and positively with effector memory subsets. This relationship lacked in HIV-infected, untreated individuals and was partially restored in the course of cART. Data are presented as median, interquartile range (IQR) for continuous variables. Changes in study parameters over time in HIV-infected subjects introducing cART were measured by Wilcoxon signed rank test; comparisons between HIV-infected and uninfected individuals were assessed by Mann-Whitney test. Tscm, T stem cell memory cells. Correlations were analyzed by Spearman’s Correlation test.</p
<p>DP: Double Positive; BD: Bone Disease; CD: Cardiovascular Disease; DN: Double Negative. MSM: Males Who Have Sex With Males. IVD: Intravenous Drug. HCV: Hepatitis C Virus. HAART: Highly Active Antiretroviral Therapy; PI: Protease... more
<p>DP: Double Positive; BD: Bone Disease; CD: Cardiovascular Disease; DN: Double Negative. MSM: Males Who Have Sex With Males. IVD: Intravenous Drug. HCV: Hepatitis C Virus. HAART: Highly Active Antiretroviral Therapy; PI: Protease Inhibitor; NNRTI: Non-Nucleoside Retroscriptase Inhibitor. DXA: Dual-energy X-ray absorptiometry. IMT: Intima Media Thickness. Data presented as: median (interquartile range, IQR) for continuous variables; absolute number (percentage) for categorical variables. p<0.05: <sup>a</sup>DP vs DN; <sup>b</sup>DP vs BD; <sup>c</sup>BD vs DN; <sup>d</sup>BD vs CD; <sup>e</sup>CD vs DN; <sup>f</sup>CD vs DP.</p><p>Patient characteristics.</p
<p>Demographic and clinical characteristics of the study population.</p
<p>CD4+ T-cell subsets expressing CCR6+CD161+ significantly increased in the peripheral blood following antiretroviral therapy, while those expressing CCR9+α4β7+ decreased upon cART. Pre-cART viral load and CD4+ T-cell activation... more
<p>CD4+ T-cell subsets expressing CCR6+CD161+ significantly increased in the peripheral blood following antiretroviral therapy, while those expressing CCR9+α4β7+ decreased upon cART. Pre-cART viral load and CD4+ T-cell activation both positively correlated with gut-homing CD4+ T-cells. Data are presented as median, interquartile range (IQR). Changes in study parameters in HIV-infected subjects introducing cART were measured by Wilcoxon signed rank test; comparisons between HIV-infected and uninfected individuals were assessed by Mann-Whitney test. Correlations were analyzed by Spearman’s Correlation test.</p
<p>No significant variation in microbial translocation and gut inflammation parameters was measured in the course of the study, while a selective increase of Lactobacillus and Bacteroides was detected among fecal bacteria genera.... more
<p>No significant variation in microbial translocation and gut inflammation parameters was measured in the course of the study, while a selective increase of Lactobacillus and Bacteroides was detected among fecal bacteria genera. LPS, lipopolysaccharide; sCD14, soluble CD14; EndocAb, Endocore toxin Antibodies. Data are presented as median, interquartile range (IQR) and were analyzed by Wilcoxon signed rank test.</p