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. 2020 Nov 28;23(12):101876. doi: 10.1016/j.isci.2020.101876

Figure 5.

Figure 5

Expanded Pa MR1T Cells Produce TNF and IL-17 following Stimulation with Agonist MR1 Ligands.

(A–D) (A) Representative flow cytometry plots of (B) TNF and (C) IL-17 expression by Pa MR1T cells, Pa non-MR1T cells, or hMAIT cells following 8 h co-culture or (D) at indicated time points of expanded (day 15–17) Pa BM MNC or hMAIT cells with freshly-thawed, autologous resting Pa BM MNC or hPBMC, respectively, that had been pulsed with MR1 ligands Ac-6-FP or 5-OP-RU, or E. coli strains EC120S, 1100-2 (both RibA+), or BSV18 (RibA-) (n = 4-7 (B and C), 2–6 (D)). PMA/ionomycin treatment for 8 h on expanded Pa BM MNC alone were used as positive controls.

(E) Flow cytometry plots and expression of IL-17 in TNF− and TNF+ Pa MR1T cells co-cultured with freshly-thawed, autologous resting Pa BM MNC pre-pulsed with 5-OP-RU (n = 6).

(F) Histograms and levels of TNF (mean fluorescence intensity; MFI) within IL-17- TNF+ and IL-17+ TNF+ Pa MR1T cell subsets following co-culture with freshly thawed, autologous resting Pa BM MNC pre-pulsed with 5-OP-RU (n = 6). See also Figures S3E–S3J. Box and whisker plot shows all data points, median, and the interquartile range. Data presented as a line graph with error bars represent the mean and standard error. Statistical significance was determined using the Mann-Whitney test (B and C), Wilcoxon's signed-rank test (E), or paired t test (F). ∗∗p < 0.01,∗p < 0.05, [∗] p < 0.1. ns, not significant.