Extended Data Fig. 2: Acquisition of cytotoxic properties by IELs requires IL-15 expression in both the lamina propria and epithelium. | Nature

Extended Data Fig. 2: Acquisition of cytotoxic properties by IELs requires IL-15 expression in both the lamina propria and epithelium.

From: IL-15, gluten and HLA-DQ8 drive tissue destruction in coeliac disease

Extended Data Fig. 2

aj, DQ8-Dd-IL-15tg, DQ8-villin-IL-15tg and DQ8-Dd-villin-IL-15tg mice were maintained on a GFD or fed gluten for 30 days (gluten). ai, The intestinal epithelium was isolated and analysed by flow cytometry. A subset of IELs was identified as TCRβ+CD4CD8αβ+ cells. In parallel, IELs were identified as TCRβ+CD4CD8+ cells by flow cytometry and quantified among IECs on H&E-stained ileum sections. a, Percentage of NKG2D+NKG2CD8αβ+ IELs are indicated (DQ8-Dd-IL-15tg sham, n = 11, gluten n = 14; DQ8-villin-IL-15tg sham, n = 20, gluten n = 20; DQ8-Dd-villin-IL-15tg sham, n = 17, gluten n = 22). b, Numbers of NKG2D+NKG2CD8+ IELs per 100 IECs (DQ8-Dd-IL-15tg sham, n = 11, gluten n = 13; DQ8-villin-IL-15tg sham, n = 20, gluten n = 20; DQ8-Dd-villin-IL-15tg sham, n = 16, gluten n = 19). c, Percentage of CD94+NKG2ACD8αβ+ IELs are indicated (DQ8-Dd-IL-15tg sham, n = 10, gluten n = 10; DQ8-villin-IL-15tg sham, n = 11, gluten n = 11; DQ8-Dd-villin-IL-15tg sham, n = 9, gluten n = 10). d, Numbers of CD94+NKG2ACD8+ IELs per 100 IECs (DQ8-Dd-IL-15tg sham, n = 10, gluten n = 10; DQ8-villin-IL-15tg sham, n = 11, gluten n = 11; DQ8-Dd-villin-IL-15tg sham, n = 9, gluten n = 10). e, Percentage of intracellular granzyme B+ IELs (DQ8-Dd-IL-15tg sham, n = 9, gluten n = 10; DQ8-villin-IL-15tg sham, n = 14, gluten n = 13; DQ8-Dd-villin-IL-15tg sham, n = 12, gluten n = 18). f, Numbers of granzyme B+ IELs per 100 IECs (DQ8-Dd-IL-15tg sham, n = 11, gluten n = 13; DQ8-villin-IL-15tg sham, n = 20, gluten n = 20; DQ8-Dd-villin-IL-15tg sham, n = 16, gluten n = 19). g, Mean fluorescent intensity (MFI) of intracellular granzyme B was measured. (DQ8-Dd-IL-15tg sham, n = 9, gluten n = 9; DQ8-villin-IL-15tg sham, n = 14, gluten n = 13; DQ8-Dd-villin-IL-15tg sham, n = 10, gluten n = 15). h, Percentage of CD8αβ+CD107a+ IELs. i, Numbers of CD8+CD107a+ IELs per 100 IECs (DQ8-Dd-IL-15tg sham, n = 7, gluten n = 6; DQ8-Dd-villin-IL-15tg sham, n = 8, gluten n = 5; DQ8-Dd-villin-IL-15tg sham, n = 5, gluten n = 4). j, Expression of Prf1 in the intestinal epithelium was measured by qPCR. Relative expression levels in sham and gluten-fed mice for each strain are shown (DQ8-Dd-IL-15tg sham, n = 16, gluten n = 16; DQ8-villin-IL-15tg sham, n = 15, gluten n = 15; DQ8-Dd-villin-IL-15tg sham, n = 20, gluten n = 26). Data are mean ± s.e.m. (j) or mean values (ag, i) from six (a, b), three (c, d, j), five (e, f), four (g) or two (h, i) independent experiments. P values were determined by one-way ANOVA with Tukey’s multiple comparison test (ag, i) or unpaired, two-tailed, t-test (j).

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