MAIT cell plasticity enables functional adaption that drives anti-bacterial immune protection

Single Cell RNA Sequencing was used to describe temporal transcriptomic changes on MAIT cells upon F.tularensis infection. We find that MAIT17-derived MAIT1 cells were distinct from canonical MAIT1 cells could migrate out of mucosal tissues to contribute to the global MAIT1 pool in subsequent systemic infections Overall design: MAIT cells were sorted (BD FACS Aria III) from pooled lungs of IL-17a-fm mice intranasally infected with 80 CFU F. tularensis LVS at 6 (4 mice), 9 (2 mice), 13 (2 mice) or 60 days (4 mice) after infection. Cells were then incubated with TotalSeqTM-C mouse hashtags for 20 minutes on ice and washed three times with PBS containing 2% BSA and resuspended in PBS containing 0.04% BSA. Approximately 7,000 cells from each sample were pooled together and loaded onto a Chromium controller (10x Genomics) for generation of gel bead-in-emulsions using 5' gene expression kits (v2; 10x Genomics). cDNA libraries were generated according to the manufacturer's instructions (10x Genomics) and then converted using the MGIEasy Universal Library Conversion Kit (BGI) before sequencing on a MGISEQ-2000 instrument (BGI).