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

Figure 2.

Figure 2

Pa MR1T Cells Specifically Proliferate to Riboflavin-Related Metabolite MR1 ligands.

(A and B) (A) Identification of Pa MR1T cells and (B) representative histograms of CellTrace Violet (CTV) dilution after 5 days in vitro culture with the MR1 ligands 5-OP-RU and Ac-6-FP, as well as riboflavin synthesis-autotroph E. coli RibA+ 1100-2 and riboflavin synthesis-deficient E. coli RibA− BSV18 in the presence of recombinant human (rh)IL-2 and IL-7 or control medium.

(C and D) (C) Expansion and (D) proliferation indices of Pa MR1T cells and non-MR1T cells following 5 days culture with various MR1 ligands and E. coli RibA+ EC120S and 1100-2 and E. coli RibA− BSV18 based on CTV dilution using the FlowJo v 9.9 software (n = 7-13). See also Figures S2G and S2H. Box and whisker plots show all data points, median, and the interquartile range. Statistical significance was determined using the Mann-Whitney test for comparisons between control medium and medium supplemented with rhIL-2+IL-7, and between Ac-6-FP and 5-OP-RU (C and D). The Wilcoxon's signed-rank test was used to determine significance between E. coli RibA+ and RibA− strains (C and D). ∗∗∗∗p < 0.0001, ∗∗∗p < 0.001,∗∗p < 0.01,∗p < 0.05. ns, not significant. iCD3, intracellular CD3.