Figure 5. MR1-restricted MAIT cell clones display ligand discrimination for the vitamin B metabolite RL-6,7-diMe. (A) A549 cells (20,000 per well) were pulsed with 5 µg/ml RL-6,7-diMe overnight and tested for their ability to stimulate MR1-restricted clones (10,000 per well) in IFN-γ ELISPOT assays. Clones D426 B1 and D481 A9 express TRAJ33; clones D481 F12 and D481 C7 express TRAJ20. TRBV usage and CDR3α and CDR3β sequences are indicated. Error bars represent SEM of duplicate determinations. (B) A549 cells (25,000 per well) were infected with S. typhimurium, C. albicans, or M. smegmatis at an MOI of 50 (hi), 25 (med), or 12.5 (lo) and then incubated overnight with the indicated MR1-restricted MAIT cell clones (10,000 per well) in duplicate wells. IFN-γ production was quantified by ELISPOT. Similar results were obtained from a minimum of three independent experiments.
Figure 5.

MR1-restricted MAIT cell clones display ligand discrimination for the vitamin B metabolite RL-6,7-diMe. (A) A549 cells (20,000 per well) were pulsed with 5 µg/ml RL-6,7-diMe overnight and tested for their ability to stimulate MR1-restricted clones (10,000 per well) in IFN-γ ELISPOT assays. Clones D426 B1 and D481 A9 express TRAJ33; clones D481 F12 and D481 C7 express TRAJ20. TRBV usage and CDR3α and CDR3β sequences are indicated. Error bars represent SEM of duplicate determinations. (B) A549 cells (25,000 per well) were infected with S. typhimurium, C. albicans, or M. smegmatis at an MOI of 50 (hi), 25 (med), or 12.5 (lo) and then incubated overnight with the indicated MR1-restricted MAIT cell clones (10,000 per well) in duplicate wells. IFN-γ production was quantified by ELISPOT. Similar results were obtained from a minimum of three independent experiments.

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