Fig 2: Reduced level of CD71 and NKG2D expression on NK cells in mice on a gluten-free (GF) diet. (a) Representative plot of lymphocyte, singlet and live gate (upper panel). Examples of dot-plots showing the gating and percentages of DX5+ cells in BALB/c standard diet (STD) and BALB/c GF mice and histograms showing expression of CD71 and NKG2D on DX5+ cells. (b) Bars represent percentages of DX5+ cells, and geometric mean fluorescence intensity (MFI) of CD71 and NKG2D gated on DX5+ cells in BALB/c and non-obese diabetic (NOD) spleen (S). Data are represented as mean values ± standard deviation of three independent experiments with three mice in each group. Black bars: STD diet; white bars: GF diet. (c) The same representation as in (b) in pancreatic lymph nodes (PLN); (d) the same representation as in (b) in auricular lymph nodes (ALN). *P < 0·05; **P < 0·01; ***P < 0·001.

Fig 3: Bystander potential of Va3.2+ CD8+ T cells. (A) Proliferation of Va3.2+ and Va3.2- naïve CD8+ T cells from Themis –/– and Themis +/+ mice in response to IL-15 stimulation. (B) Histogram summary of the proliferative responses to IL-15 stimulation. (C) Proportion of Va3.2+ and non-Va3.2+ CD8 T cells upon IL-15 stimulation. (D) Proportions of Va3.2+ T cells in the bystander (CD8+ Tetramerneg CD44hi NKG2D+) and control (CD8+ Tetramerneg CD44lo) population on day 4 of LM-OVA infection in uninfected and infected Themis –/– and Themis +/+ mice. Data are representative from three independent experiments with 3-4 biological replicates per genotype per experiment. nsnot significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 as determined by two-sided Student’s t-test. All error bars represent SDs.

Fig 4: Reduced mRNA levels of NKG2D and DX5 in gluten-free (GF) mice. (a) Relative messenger RNA expression levels of NKG2D and DX5 in tissues from 13-week-old mice kept on a gluten free (GF), strictly gluten-free (SGF) or matched controls, standard diet (STD) and strictly standard diet (SSTD). Expression levels of NKG2D and DX5 in intestinal tissue (upper graphs). NKG2D and DX5 expression levels in islets (lower graphs). (b) Relative messenger RNA levels of NKG2D and DX5 in intestines from SGF-fed non-obese diabetic (NOD) mice at 8, 13 and 20 weeks of age. *P < 0·05; **P < 0·01; ***P < 0·001.

Fig 5: Reduced level of CD71 expression and CD8+ cells in mice on a gluten-free (GF) diet. (a) Representative plot of lymphocyte, singlet and live gate (upper panel). Examples of dot-plots showing the gating and percentages of CD8+ cells in BALB/c standard diet (STD) and BALB/c GF mice and histograms showing expression of CD71 and NKG2D on CD8+ cells. (b) Bars represent percentages of CD8+ cells, and geometric mean fluorescence intensity (MFI) of CD71 and NKG2D gated on CD8+ cells in BALB/c and non-obese diabetic (NOD) spleen (S). Data are represented as mean values ± standard deviation of three independent experiments with three mice in each group. Black bars: STD diet; white bars: GF diet. (c) The same representation as in (b) in pancreatic lymph nodes (PLN); (d) the same representation as in (b) in auricular lymph nodes (ALN). *P < 0·05; **P < 0·01; ***P < 0·001.