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. 2020 Nov 20;11:585134. doi: 10.3389/fimmu.2020.585134

Figure 5.

Figure 5

Optimal IL-17A and Il-17F production from MAIT cells requires MR1 and IL-12/IL-18 signaling. (A) Representative bivariate FC plots of IL-17F and IL-17A production by MAIT cells following culture of PBMCs with fixed E. coli for 1, 2, or 3 days with IL-12 and IL-18. (B) Frequency of cytokine producing MAIT cells following 1, 2, or 3 days of PBMC culture with fixed E. coli, IL-12, and IL-18. Significance calculated using two-way ANOVA with multiple comparisons across days (n = 8). (C) Median fluorescence intensity of cytokine producing MAIT cells cultured as in (B), analyzed as the median fluorescence intensity gated on the cytokine producing subset. Significance calculated using two-way ANOVA with multiple comparisons across days (n = 8). (D–H) Combined results showing the frequency of cytokine producing CD8+ MAIT cells following 3 days of culture with fixed E. coli with or without IL-12 and IL-18, and with or without the addition of antibodies inhibiting MR1 or IL-23p19. Statistics calculated as two-way ANOVA, with repeated measures using either ‘E. coli’ or ‘IL-12/IL-18 E. coli’ as reference group (n = 10). (I) Representative bivariate FC plots of PBMCs stimulated with fixed E. coli, IL-12 and IL-18 for 72 h, showing cytokine production gated on total CD3+ T cells, with or without MR1 blockade. **p<0.01, ****p<0.0001.