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MAIT Cells pp 23-38 | Cite as

Isolation and Characterization of MAIT Cells from Human Tissue Biopsies

  • Daniela Di Blasi
  • Alessandro Vacchini
  • Gennaro De Libero
  • Lucia MoriEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2098)

Abstract

Human mucosal-associated invariant T (MAIT) cells are unconventional T cells highly enriched in tissues exposed to microbial antigens including the oral, gastrointestinal and genital mucosae, liver, and lung. Here we describe a protocol for isolation and characterization of peripheral blood and tissue-infiltrating MAIT cells by using multicolor flow cytometry. This technology allows the analysis of multiple markers in a single sample at a single-cell level. Study of human samples requires particular care since the sample amount is often limited. We present a protocol optimized for the isolation and characterization of human MAIT cells and the identification of MAIT cell populations detected by simultaneous expression of multiple activation markers and inhibitory receptors.

Key words

Human MR1 tetramer Immune phenotype MAIT cells Multicolor flow cytometry Tissue-resident cells 

Notes

Acknowledgments

We thank Paula Cullen and Julian Spagnuolo for critical reading of the manuscript, and the DBM Flow Facility staff. The MR1 tetramer technology was developed jointly by Dr. James McCluskey, Dr. Jamie Rossjohn, and Dr. David Fairlie. The MR1 tetramer was produced by the NIH Tetramer Core Facility as permitted to be distributed by the University of Melbourne. The author’s studies are supported by SNF 310030-173240, KFS-3730-08-2015, and PMB-02-17 Grants.

References

  1. 1.
    Tilloy F, Treiner E, Park SH, Garcia C, Lemonnier F, de la Salle H, Bendelac A, Bonneville M, Lantz O (1999) An invariant T cell receptor alpha chain defines a novel TAP-independent major histocompatibility complex class Ib-restricted alpha/beta T cell subpopulation in mammals. J Exp Med 189:1907–1921CrossRefGoogle Scholar
  2. 2.
    Treiner E, Duban L, Bahram S, Radosavljevic M, Wanner V, Tilloy F, Affaticati P, Gilfillan S, Lantz O (2003) Selection of evolutionarily conserved mucosal-associated invariant T cells by MR1. Nature 422:164–169CrossRefGoogle Scholar
  3. 3.
    Gold MC, Cerri S, Smyk-Pearson S, Cansler ME, Vogt TM, Delepine J, Winata E, Swarbrick GM, Chua WJ, Yu YY, Lantz O, Cook MS, Null MD, Jacoby DB, Harriff MJ, Lewinsohn DA, Hansen TH, Lewinsohn DM (2010) Human mucosal associated invariant T cells detect bacterially infected cells. PLoS Biol 8:e1000407CrossRefGoogle Scholar
  4. 4.
    Tang XZ, Jo J, Tan AT, Sandalova E, Chia A, Tan KC, Lee KH, Gehring AJ, De Libero G, Bertoletti A (2013) IL-7 licenses activation of human liver intrasinusoidal mucosal-associated invariant T cells. J Immunol 190:3142–3152CrossRefGoogle Scholar
  5. 5.
    Lepore M, Kalinichenko A, Colone A, Paleja B, Singhal A, Tschumi A, Lee B, Poidinger M, Zolezzi F, Quagliata L, Sander P, Newell E, Bertoletti A, Terracciano L, De Libero G, Mori L (2014) Parallel T-cell cloning and deep sequencing of human MAIT cells reveal stable oligoclonal TCRbeta repertoire. Nat Commun 5:3866CrossRefGoogle Scholar
  6. 6.
    Kjer-Nielsen L, Patel O, Corbett AJ, Le Nours J, Meehan B, Liu L, Bhati M, Chen Z, Kostenko L, Reantragoon R, Williamson NA, Purcell AW, Dudek NL, McConville MJ, O’Hair RA, Khairallah GN, Godfrey DI, Fairlie DP, Rossjohn J, McCluskey J (2012) MR1 presents microbial vitamin B metabolites to MAIT cells. Nature 491:717–723CrossRefGoogle Scholar
  7. 7.
    Miley MJ, Truscott SM, Yu YY, Gilfillan S, Fremont DH, Hansen TH, Lybarger L (2003) Biochemical features of the MHC-related protein 1 consistent with an immunological function. J Immunol 170:6090–6098CrossRefGoogle Scholar
  8. 8.
    Huang S, Gilfillan S, Kim S, Thompson B, Wang X, Sant AJ, Fremont DH, Lantz O, Hansen TH (2008) MR1 uses an endocytic pathway to activate mucosal-associated invariant T cells. J Exp Med 205:1201–1211CrossRefGoogle Scholar
  9. 9.
    Treiner E, Duban L, Moura IC, Hansen T, Gilfillan S, Lantz O (2005) Mucosal-associated invariant T (MAIT) cells: an evolutionarily conserved T cell subset. Microbes Infect 7:552–559CrossRefGoogle Scholar
  10. 10.
    Gold MC, McLaren JE, Reistetter JA, Smyk-Pearson S, Ladell K, Swarbrick GM, Yu YY, Hansen TH, Lund O, Nielsen M, Gerritsen B, Kesmir C, Miles JJ, Lewinsohn DA, Price DA, Lewinsohn DM (2014) MR1-restricted MAIT cells display ligand discrimination and pathogen selectivity through distinct T cell receptor usage. J Exp Med 211:1601–1610CrossRefGoogle Scholar
  11. 11.
    Reantragoon R, Kjer-Nielsen L, Patel O, Chen Z, Illing PT, Bhati M, Kostenko L, Bharadwaj M, Meehan B, Hansen TH, Godfrey DI, Rossjohn J, McCluskey J (2012) Structural insight into MR1-mediated recognition of the mucosal associated invariant T cell receptor. J Exp Med 209:761–774CrossRefGoogle Scholar
  12. 12.
    Dusseaux M, Martin E, Serriari N, Peguillet I, Premel V, Louis D, Milder M, Le Bourhis L, Soudais C, Treiner E, Lantz O (2011) Human MAIT cells are xenobiotic-resistant, tissue-targeted, CD161hi IL-17-secreting T cells. Blood 117:1250–1259CrossRefGoogle Scholar
  13. 13.
    Reantragoon R, Corbett AJ, Sakala IG, Gherardin NA, Furness JB, Chen Z, Eckle SB, Uldrich AP, Birkinshaw RW, Patel O, Kostenko L, Meehan B, Kedzierska K, Liu L, Fairlie DP, Hansen TH, Godfrey DI, Rossjohn J, McCluskey J, Kjer-Nielsen L (2013) Antigen-loaded MR1 tetramers define T cell receptor heterogeneity in mucosal-associated invariant T cells. J Exp Med 210:2305–2320CrossRefGoogle Scholar
  14. 14.
    Corbett AJ, Eckle SB, Birkinshaw RW, Liu L, Patel O, Mahony J, Chen Z, Reantragoon R, Meehan B, Cao H, Williamson NA, Strugnell RA, Van Sinderen D, Mak JY, Fairlie DP, Kjer-Nielsen L, Rossjohn J, McCluskey J (2014) T-cell activation by transitory neo-antigens derived from distinct microbial pathways. Nature 509:361–365CrossRefGoogle Scholar
  15. 15.
    Schmaler M, Colone A, Spagnuolo J, Zimmermann M, Lepore M, Kalinichenko A, Bhatia S, Cottier F, Rutishauser T, Pavelka N, Egli A, Azzali E, Pieroni M, Costantino G, Hruz P, Sauer U, Mori L, De Libero G (2018) Modulation of bacterial metabolism by the microenvironment controls MAIT cell stimulation. Mucosal Immunol 11:1060–1070CrossRefGoogle Scholar
  16. 16.
    Di Blasi D, Boldanova T, Mori L, Terracciano L, Heim MH, De Libero G (2019) Cell Mol Gastroenterol Hepatol,  https://doi.org/10.1016/j.jcmgh.2019.08.004
  17. 17.
    van der Maaten L (2014) Accelerating t-SNE using tree-based algorithms. J Mach Learn Res 15:3221–3245Google Scholar
  18. 18.
    Lepore M, Kalinichenko A, Calogero S, Kumar P, Paleja B, Schmaler M, Narang V, Zolezzi F, Poidinger M, Mori L, De Libero G (2017) Functionally diverse human T cells recognize non-microbial antigens presented by MR1. Elife 6:e24476CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Daniela Di Blasi
    • 1
  • Alessandro Vacchini
    • 1
  • Gennaro De Libero
    • 1
  • Lucia Mori
    • 1
    Email author
  1. 1.Experimental Immunology, Department of BiomedicineUniversity Hospital and University of BaselBaselSwitzerland

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