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

Figure 6.

Figure 6

Expanded Pa MR1T Cells Kill Target Epithelial Cell Lines Pulsed with Agonist MR1 Ligands.

(A) Histograms and (B) levels (MFI) of the MR1-5-OP-RU tetramer staining of expanded (d15-17) Pa MR1T cells or hMAIT cells following 24 h co-culture with Pa kidney (PaKi) cell line fed with Ac-6-FP, 5-OP-RU, E. coli RibA− (BSV18), or E. coli RibA+ (1100-2 or EC120S) (n = 6-13).

(C and D) (C) Flow cytometry plots and (D) expression of active caspase (Casp)3 and amine-reactive cytoplasmic dye (dead cell marker; DCM) by the PaKi target cells fed with various antigens as indicated following 24 h co-culture with expanded (d15-17) Pa MR1T or hMAIT cells (n = 6-22).

(E and F) Correlation between frequency of Pa MR1T cells within the expanded culture and frequency of Casp3+ and/or DCM + PaKi cell lines fed with (E) 5-OP-RU or E. coli RibA+ (1100-2 or EC120S) (n = 19), or (F) Ac-6-FP or E. coli RibA− (BSV18) (n = 6-9).

(G, H, and I) Histograms and MFI of the MR1-5-OP-RU tetramer staining of expanded Pa MR1T cells or hMAIT cells (G) and 5-OP-RU-pulsed 293T-hMR1 target cells apoptosis (H and I) following 24 h co-culture, in the presence of anti-hMR1 mAb or IgG2a isotype control (n = 5-6). See also Figure S4. Box and whisker plot shows all data points, median, and the interquartile range. Statistical significance was determined using unpaired t test (B) and the Mann-Whitney test (D and I). Correlations were assessed using the Spearman rank correlation (E and F). ∗∗∗p < 0.001, ∗∗p < 0.01,∗p < 0.05. ns, not significant.