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Author: MCCLUSKEY, JAMES ×

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    Characterization and Purification of Mouse Mucosal-Associated Invariant T (MAIT) Cells
    Chen, Z ; Wang, H ; D'Souza, C ; Koay, H-F ; Meehan, B ; Zhao, Z ; Pediongco, T ; Shi, M ; Zhu, T ; Wang, B ; Kjer-Nielsen, L ; Eckle, SBG ; Rossjohn, J ; Fairlie, DP ; Godfrey, DI ; Strugnell, RA ; McCluskey, J ; Corbett, AJ (John Wiley & Sons, 2019)
    This unit describes the utility of various mouse models of infection and immunization for studying mucosal-associated invariant T (MAIT) cell immunity: MAIT cells can be isolated from the lungs (or from other tissues/organs) and then identified and characterized by flow cytometry using MR1 tetramers in combination with a range of antibodies. The response kinetics, cytokine profiles, and functional differentiation of lung MAIT cells are studied following infection with the bacterial pathogen Legionella longbeachae or Salmonella enterica Typhimurium or immunization with synthetic MAIT cell antigen plus Toll-like receptor agonist. MAIT cells enriched or expanded during the process can be used for further studies. A step-by-step protocol is provided for MAIT cell sorting and adoptive transfer. Mice can then be challenged and MAIT cells tracked and further examined.
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    Human blood MAIT cell subsets defined using MR1 tetramers
    Gherardin, NA ; Souter, MNT ; Koay, H-F ; Mangas, KM ; Seemann, T ; Stinear, TP ; Eckle, SBG ; Berzins, SP ; d'Udekem, Y ; Konstantinov, IE ; Fairlie, DP ; Ritchie, DS ; Neeson, PJ ; Pellicci, DG ; Uldrich, AP ; McCluskey, J ; Godfrey, DI (WILEY, 2018-05)
    Mucosal-associated invariant T (MAIT) cells represent up to 10% of circulating human T cells. They are usually defined using combinations of non-lineage-specific (surrogate) markers such as anti-TRAV1-2, CD161, IL-18Rα and CD26. The development of MR1-Ag tetramers now permits the specific identification of MAIT cells based on T-cell receptor specificity. Here, we compare these approaches for identifying MAIT cells and show that surrogate markers are not always accurate in identifying these cells, particularly the CD4+ fraction. Moreover, while all MAIT cell subsets produced comparable levels of IFNγ, TNF and IL-17A, the CD4+ population produced more IL-2 than the other subsets. In a human ontogeny study, we show that the frequencies of most MR1 tetramer+ MAIT cells, with the exception of CD4+ MAIT cells, increased from birth to about 25 years of age and declined thereafter. We also demonstrate a positive association between the frequency of MAIT cells and other unconventional T cells including Natural Killer T (NKT) cells and Vδ2+ γδ T cells. Accordingly, this study demonstrates that MAIT cells are phenotypically and functionally diverse, that surrogate markers may not reliably identify all of these cells, and that their numbers are regulated in an age-dependent manner and correlate with NKT and Vδ2+ γδ T cells.