IL-17 producing cell function is higher in colorectum than blood in uninfected RMs.

<p><b>(A)</b> Comparison between cytokine profiles of Th17, Th22, and Th17/Th22 cells in PBMC and RB in uninfected RMs (d. -20 p.i.). Cytokine profiles were generated for each cell population by SPICE program v. 5.33, and were calculated by Flowjo Boolean gating. <b>(B)</b> Functional scores were compared for all three subsets between PBMC and RB. Functional scores represent average number of cytokines produced per individual cell (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005412#sec011" target="_blank">Methods</a>). Averaged data are presented as means ± SEM.</p

Loss of Th17, Th22, and Th17/Th22 cell function and levels are correlates and predictors of SIV persistence.

<p>Correlations between colorectal SIV-DNA content at late-ART (d. 256 p.i.) and IL-17 and IL-22 producing cell function and levels at both ART (<b>A-C</b>) and pre-ART infection (d. 58 p.i). <b>(D,E)</b>. Levels and function of the three subsets at pre-ART and after ART interruption additionally correlated with levels of blood DNA levels after ART interruption (d. 440 p.i.) <b>(F-I)</b>. Pearson product-moment correlation coefficients were measured for all plots except for 8E, which required Spearman’s rank correlation coefficient.</p

SIV infection severely ablates intestinal IL-17 and IL-22 producing cell function and levels.

<p><b>(A)</b> Comparison between PBMC and RB Th17, Th22, and Th17/Th22 cell cytokine profiles at pre-infection (d.-20 p.i.) vs SIV infection (d. 58 p.i.). While all RB cytokine profiles significantly changed after SIV infection, the blood cytokine profiles remained unchanged. <b>(B)</b> No changes in functional score in PBMC after SIV infection. <b>(C)</b> In all three subsets, colorectal functional scores drastically decreased after SIV infection. Th17 cells marked as red, Th22 cells marked as blue, and Th17/Th22 cells marked as green. Averaged data are presented as means ± SEM.</p

Loss of Th17, Th22, and Th17/Th22 cell function correlates with colorectal immune activation, soluble markers of inflammation, and levels of CD4⁺ T-cells.

<p>Correlations between Th17, Th22, and Th17/Th22 cell function and plasma viral loads (copies viral RNA per mL of plasma) <b>(A,B)</b>, absolute number of CD4⁺ T-cells <b>(C)</b>, levels of activated colorectal (HLA-DR⁺CD38⁺) CD4⁺ T-cells <b>(D,E)</b>, the level of proliferating (Ki-67⁺) RB CD4⁺ T-cells <b>(F)</b>, as well as soluble markers of inflammation sCD14 and sCD163 <b>(G, H)</b>. Pearson product-moment correlation coefficients were measured for all plots except for 7A, which required Spearman’s rank correlation coefficient.</p

Levels of Th17, Th22, and Th17/Th22 are more drastically depleted by SIV infection in RB than PBMC.

<p>Comparison of frequencies (measured as percentages of total CD4⁺ T-cell populations) of circulating <b>(A)</b> and colorectal <b>(B)</b> Th17 (red), Th22 (blue), and Th17/Th22 (green) cells before (d.-20 p.i.) and after (d. 58 p.i.) SIV infection. Averaged data are presented as means ± SEM.</p

Study timeline and representative cytokine panel staining.

<p><b>(A)</b> Complete study timeline of longitudinal tissue collections, SIVmac239 i.v. infection (d.0 p.i.), ART treatment (d. 58 to d.256 p.i.), and off-ART time period. <b>(B)</b> Representative staining for IL-17, IL-22, IFNγ, TNFα, and IL-2 within intestinal CD4⁺ T-cells in a representative uninfected RM.</p

ART treatment is insufficient for full restoration of cell subset levels and function.

<p><b>(A-C)</b> Longitudinal functional scores of Th17, Th22, and Th17/Th22 cells in RB at pre-infection (d. -20 p.i.), pre-ART (d. 58 p.i.), and throughout ART (d. 84 p.i. through d. 256 p.i.). Data are presented as box and whisker plots, with the median functional score plotted in between the 25% and 75% quartiles. Dotted line marks time of SIV infection and shaded gray box represents time of ART treatment. ART significantly increased all three subsets’ functional scores, but did not bring Th17 cell function back to pre-infection level. <b>(D)</b> Levels of subsets (as percentages of total CD4⁺ T-cell populations) in RB during ART remain significantly lower than pre-infection. <b>(E)</b> Longitudinal cumulative subset scores, calculated by multiplying cell frequencies and functional scores, showed the inability of ART to fully restore the levels and function of Th17, Th22, and Th17/Th22 cells. Th17 cells marked as red, Th22 cells marked as blue, and Th17/Th22 cells marked as green. Averaged data are presented as means ± SEM.</p

Dufour et al. Source Data.xlsx

Dataset used to create every graph of the paper "Phenotypic characterization of single CD4+ T cells harboring genetically intact and inducible HIV genomes" in Nature Communications Author list: Caroline Dufour1, Corentin Richard1, Marion Pardons1, Marta Massanella1, Antoine Ackaoui1, Ben Murrell2, Bertrand Routy1, Réjean Thomas3, Jean-Pierre Routy4, Rémi Fromentin1, Nicolas Chomont1 1Centre de Recherche du CHUM and Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, H2X 0A9, Quebec, Canada 2Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, 171 77, Sweden 3Clinique médicale l’Actuel, Montreal, H2L 4P9, Quebec, Canada 4Division of Hematology & Chronic Viral Illness Service, McGill University Heath Centre, Montreal, H4A 3J1, Quebec, Canada</p

Ehretia dicksonii Hance var. japonica Nakai

原著和名: マルバチシャノキ科名: ムラサキ科 = Boraginaceae採集地: 千葉県 安房郡 天津小湊町 実入 (安房 天津小湊町 実入)採集日: 1966/6/26採集者: 萩庭丈壽整理番号: JH001226国立科学博物館整理番号: TNS-VS-95122

Co-expression of PD-1, TIGIT and LAG-3 is a marker of HIV persistence during ART.

<p>(A) Frequency of CD4⁺ T cells co-expressing PD-1 and/or TIGIT and/or LAG-3 (PD-1/TIGIT/LAG-3 triple–(P-T-L-), PD-1 single + (P+), TIGIT single + (T+), LAG-3 single + (L+), PD-1/TIGIT double + (P+T+), TIGIT/LAG-3 double + (T+L+), PD-1/LAG-3 double + (P+L+) and PD-1/TIGIT/LAG-3 triple + (P+T+L+)) determined by Boolean gating in cohort 1 (n = 48). Horizontal bars indicate median values with interquartile ranges. (B) Venn diagram showing the pattern of co-expression of PD-1, TIGIT and LAG-3. (C), (D), (E), (F) Associations between the frequency of CD4⁺ T cells harboring integrated HIV DNA and the frequency of CD4⁺ T cells expressing none of these markers (triple–), PD-1 and TIGIT (double +), TIGIT and LAG-3 (double +) and PD-1 and TIGIT and LAG-3 (triple +), respectively. P values were obtained from negative binomial regression analysis. Effect sizes for the associations are as follows: (C) 0.69-fold-change in integrated HIV DNA for 1 point increase in percentage of PD-1/TIGIT/LAG-3 triple—CD4⁺ T cells, (D) 1.18-fold-change in integrated HIV DNA for 1 point increase in percentage of PD-1/TIGIT double + CD4⁺ T cells, (E) 1.30-fold-change in integrated HIV DNA for 1 point increase in percentage of TIGIT/LAG-3 double + CD4⁺ T cells and (F) 1.94-fold-change in integrated HIV DNA for 1 point increase in percentage of PD-1/TIGIT/LAG-3 triple + CD4⁺ T cells. Open circles represent the limit of detection in the negative samples (based on cell input).</p