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
Part of the Methods in Molecular Biology book series (MIMB, volume 2098)


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 



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.


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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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