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 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).