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. 2022 Aug 1;11(8):e1407. doi: 10.1002/cti2.1407

Figure 2.

Figure 2

Peripheral T cells from Crohn's disease (CD) donors have a more inflammatory phenotype than those from non‐inflammatory bowel disease (non‐IBD) donors. PBMCs were isolated from non‐IBD or CD donors. (a) Frequency of PBMC CD4+IL‐17A+, CD4+IFN‐γ+ and regulatory T cells (CD4+CD25+FOXP3hi) of CD3+ cells. Data are shown as individual patients, and the bar represents the median frequency. Non‐IBD (n = 9), CD (n = 8). (b) 80 000 PBMCs were cultured with anti‐CD3/anti‐CD28 beads at a 1:1 ratio for 48 h. Cytokine supernatants were detected using a LegendPlex Human T Cell Cytokine Panel, then analysed by flow cytometry. Data are shown as individual patients, and the bar represents the median concentration (pg mL−1). Where appropriate, upper or lower limits of detection are shown with the dotted line. Non‐IBD (n = 7), CD (n = 10). (c) Tconvs were stained with Cell‐Trace Violet to allow for tracking of division events. Precursor frequency was calculated by (N/2 i )/(Total number of events/2 i ) × 100 = % P, then precursor frequency determined by the sum of % P of all peaks where division occurred (peaks 1–4). Proliferation index was calculated by ((N/2 i ) × N)/(Total number of events), where N indicates the number of events within a proliferation peak and 2 i indicates the number progeny per division stage. Non‐IBD (n = 6), CD (n = 6). Statistical analyses were calculated using the Mann–Whitney U‐test (*P < 0.05, **P < 0.01). (d) PBMCs were isolated from non‐IBD and CD donors and cultured for 72 h in the presence or absence of 107 heat‐killed Faecalibacterium prausnitzii, then analysed by flow cytometry. Left top: Frequency of CD4+IFN‐γ+ T cells. Right top: Frequency of CD4+IL‐17A+ T cells. Left bottom: Frequency of regulatory T cells (CD4+CD25+FOXP3HI). Right bottom: Frequency of CD4+CD25HIFOXP3− T cells. Statistical analyses were calculated using the Wilcoxon matched‐pairs signed‐rank test, (**P < 0.01, ***P < 0.001). Non‐IBD (n = 9), CD (n = 8). Lines represent matched‐patient PBMCs in each condition. Similar results were obtained with culture of 106 bacteria.