Single-cell analysis of human MAIT cell transcriptional, functional and clonal diversity [Exp 3]

MAIT cells are innate-like T cells that recognize microbial metabolites through a semi-invariant T cell receptor (TCR). Major questions remain regarding the extent of human MAIT cell functional and clonal diversity. To address these, we analyzed the single-cell transcriptome and TCR repertoire of blood and liver MAIT cells and developed functional RNA-sequencing (fRNA-seq), a method to integrate function and TCR clonotype at single-cell resolution. MAIT cell clonal diversity was comparable to conventional memory T cells, with private TCR repertoires shared across matched tissues. Baseline functional diversity was low and largely related to tissue site. MAIT cells showed stimulus-specific transcriptional responses in vitro, with cells positioned along gradients of activation. Clonal identity influenced resting and activated transcriptional profiles, but intriguingly was not associated with the capacity to produce IL-17. Overall, MAIT cells show phenotypic and functional diversity according to tissue localization, stimulation environment, and clonotype. Overall design: Enriched CD8+ T cells (n = 3 donors) were left unstimulated, or stimulated with MR1/5-OP-RU + aCD28 or IL-12 + IL-18. After 20 hours, MAIT cells (CD3+CD8+CD26+CD161hiVa7.2+) were sorted for single-cell RNA-sequencing (scRNA-seq) and single-cell TCR-sequencing (scTCR-seq) using 10x Genomics technology.