10X PBS pH 7.2，orb348532，Biorb

A subset of Tem-like cells sorted based on surface markers defining clusters 12 and 13 are highly susceptible to HIV infection. (A) Shown are the CyTOF datasets, with UI CD4+ T cells shown in gray and the HIV-susceptible PRE cells shown in red. Cells were pre-gated on live, singlet CD3 + CD19−CD8−CD4+ T cells. A sequential gating strategy was then implemented using surface markers characteristic of HIV-susceptible cells as defined by clusters 12 and 13. This strategy was used to characterize a final population of "population-1" cells (CD3 + CD4+ CD45RO + CD45RA−CCR7low/medCD29med/highCD69med/high CD62LlowCD57low/med), which were more abundant among PRE cells than among UI cells. For comparison, we characterized a "population-2" (CD3 + CD4+ CD45RO + CD45RA−CCR7low/medCD29lowCD69low and not CD62LlowCD57low/med) predicted to be much less susceptible to infection because it comprised a significantly lower proportion of PRE cells. The gating strategies are shown on the left, whereas the graphs on the right depict the frequencies of the population-1 and population-2 subsets within the UI and PRE cell populations. Note that the over-representation of population-1 cells among PRE cells suggest their preferential susceptibility to infection, whereas the under-representation of population-2 cells among PRE cells suggest their relative resistance to infection. *p < 0.05, ****p < 0.0001 as determined by a Student's paired t test. Error bars correspond to the standard deviation.

Quantification of CD8 and CD4 T cell clusters by CyTOF analysis in LP of control and CD patients. a Schematic t-SNE of CD4+ and CD8+ T cells from LP of all donors concatenated together (n = 18) controls (N), n = 8; CD, non-inflamed site (NI), n = 9; CD, inflamed site (II), n = 6. Total of 23 samples. b t-SNE of the indicated markers in CD4+ and CD8+ T cells. c, d Quantification of total CD8+ (c), total CD4+ (d) in LP of controls and CD patients by CyTOF (triangles = fresh samples) and FACS (circles = frozen samples). c, d Control (N), n = 17 (8 fresh, 9 frozen); CD, non-inflamed site (NI) n = 19 (9 fresh, 10 frozen); CD, inflamed site (II), n = 14 (6 fresh, 8 frozen). e–i Quantification of total CD8+ TRM (e) and CD8+ clusters 2 (f), 4 (g), 12 (h), and 16 (i) in LP of controls and CD patients by CyTOF. j–l Quantification of the CD4+ clusters 5 and 14 (j), 7 (k), and 9 (l) in LP of controls and CD patients by CyTOF. e–l Controls (N), n = 8; CD, non-inflamed site (NI), n = 9; CD, inflamed site (II), n = 6. Circles and triangles on the boxplots show data collected for each individual donor. Data were median and interquartile range. Significance was calculated using an ordinary, one-way ANOVA, multiple comparisons test with Prism v8 software. c **P = 0.0014; d **P = 0.028; e *P = 0.0139; f *P = 0.0178; h *P = 0.0178; i *P = 0.0219; j N vs. NI *P = 0.0156, NI vs. II *P = 0.0465; k **P = 0.0014; l **P = 0.0283. TRM tissue-resident memory T cell.