Fig 2: Cell subset expression in mild (n = 6) and moderate-severe (n = 19) AR patients. (A) Percentages and absolute counts of lymphocyte subsets: T cells (CD3+), B cells (CD3-CD19+), and NK cells (CD3-CD16/56+). (B) Percentages and absolute counts of T cell subsets: CD8 cells (CD3+CD8+) and CD4 cells (CD3+CD4+). (C) Percentages and absolute counts of CD4+ T cell subsets: Treg (CD25+CD127low/-), Th1 (CXCR3+CCR4-CCR6-), Th2 (CXCR3-CCR4+CCR6-), and Th17 (CXCR3-CCR4+CCR6+ and CXCR3+CCR4-CCR6+). Horizontal lines indicate mean ± SD. P-values were calculated with the two-tailed Mann-Whitney U test.

Fig 3: HIV-1 exposure alters the T cell expression of transcriptional factors and chemokine receptors.Mucosal immune cells isolated from the colorectal tissue biopsies by enzymatic digestion were exposed to HIV-1BaL (250 ng/mL), either free (F-HIV), complement-opsonized (C-HIV), or virions opsonized by a cocktail of complement and antibodies (CI-HIV) or mock-treated, by spinning the cultures. The effect HIV exposure had on mucosal T cells phenotype was assessed after 4 days. (A–D) The immune cells were stained with CD3, CD4, CD8, Tbet, GATA3, ROR?t, and FoxP3 mAbs. (A–B) Percentage of unexposed, F-HIV, C-HIV, or CI-HIV exposed CD4+ T cells expressing GATA3, Tbet, ROR?t, or FoxP3, or ROR?t and Tbet were assessed by flow cytometry. (C–D) Percentage of unexposed, F-HIV, C-HIV, or CI-HIV exposed CD8+ T cells expressing GATA3, Tbet, ROR?t, or FoxP3, or ROR?t and Tbet were assessed by flow cytometry. (E–F) The immune cells were stained with CD3, CD4, CD8, CCR4, CCR6, and CXCR3 mAbs. (E) The proportion of each of the four T helper populations: CCR4+CCR6-, CCR4+CCR6+, CXCR3+CCR6- or CXCR3+CCR6+ among unexposed CD4+ T cells were assessed by flow cytometry. (F) Percentage of unexposed, F-HIV, C-HIV, or CI-HIV exposed CD4+ T cells expressing CCR4+CCR6-, CCR4+CCR6+, CXCR3+CCR6- or CXCR3+CCR6+ were assessed by flow cytometry (N = 12–15). Statistical significance was tested using repeated measures of ANOVA followed by Tukey's posttest. *p<0.05, **p<0.01, ***p<0.001. Data are shown as mean ± SEM.

Fig 4: Cell subset expression in AR patients (n = 25) and HCs (n = 20). (A) Percentages and absolute counts of lymphocyte subsets: T cells (CD3+), B cells (CD3-CD19+), and NK cells (CD3-CD16/56+). (B) Percentages and absolute counts of T cell subsets: CD8 cells (CD3+CD8+) and CD4 cells (CD3+CD4+). (C) Percentages and absolute counts of CD4+ T cell subsets: Treg (CD25+CD127low/-), Th1 (CXCR3+CCR4-CCR6-), Th2 (CXCR3-CCR4+CCR6-), and Th17 (CXCR3-CCR4+CCR6+ and CXCR3+CCR4-CCR6+). Horizontal lines indicate mean ± SD. P-values were calculated with the two-tailed Mann-Whitney U test.

Fig 5: Representative flow cytometry gating strategy for identification of CD4+T subsets. Treg (CD25+CD127low/-), Th1 (CXCR3+CCR4−CCR6−), Th2 (CXCR3−CCR4+CCR6−), and Th17 (CXCR3−CCR4+CCR6+ and CXCR3+CCR4−CCR6+) in peripheral blood mononuclear cells. The data were obtained from 1 healthy control subject.