Extended Data Fig. 2: Validation of adoptive transfer of MAIT cells.
a, Representative flow cytometry profiles depict gating strategies to sort donor MAIT cells for adoptive transfer experiments. Meningeal tissue was pooled, with tissue from 5 mice in 1 ml of Liberase digestion buffer. Digested tissue from up to 60 mice was pooled per sample, followed by staining with MR1 tetramers and surface antibodies to identify MAIT cells. Events of around 8% of the sample (equivalent to around 5 mice) were collected in the representative flow cytometry profiles. CD3 and TCRβ antibodies were not included for purification of MAIT cells for adoptive transfer. Donor MAIT cells were identified as CD45+ Thy1.2hiIL-18RhiMR1-tetramer+cells. b, Representative profiles of MAIT cells in the meninges of 7-month-old Mr1−/− mice that received adoptive transfer of PBS or MAIT cells. Plots were pre-gated on CD45+CD11b−B220−NK1.1- Thy1.2+ cells. c, Numbers of meningeal MAIT cells per mouse in the meninges and in the small intestinal laminal propria (SILP) of recipient mice at the indicated time points after adoptive transfer. d, Percentages of MAIT cells in the total T cell subset in the dura/arachnoid meningeal tissue obtained from inner calvaria and in the leptomeninges of recipient mice at 6 months post adoptive transfer. Error bars = Mean ± SE. Data are from 3 independent experiments, 2-5 recipient mice pooled in one sample for each experiment (b–d). Each data point indicates one independent experiment (c, d).
