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Human MAIT Cell Activation In Vitro

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MAIT Cells

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

Abstract

Mucosal-associated invariant T (MAIT) cells are an abundant innate-like T cell subset in humans, enriched in mucosal tissues and the liver. MAIT cells express a semi-invariant T cell receptor (TCR) and recognize microbial-derived riboflavin metabolites presented on the MHC Class I-like molecule MR1. In addition to activation via the TCR, MAIT cells can also be activated in response to cytokines such as IL-12 and IL-18, in contrast to conventional T cells. Here we describe TCR-dependent and -independent methods for MAIT cell activation. The TCR-dependent approaches include stimulation with microbead- or plate-bound anti-CD3/anti-CD28 antibodies, and with 5-OP-RU or paraformaldehyde (PFA)-fixed E. coli in the presence of antigen-presenting cells (APCs). The latter method includes a combination of TCR- and cytokine-mediated stimulation. The TCR-independent methods include direct stimulation with the recombinant cytokines IL-12 and IL-18, and indirect stimulation with TLR-4/TLR-8 agonists or influenza A virus in the presence of APCs. Finally, we outline a protocol to analyze activated MAIT cells using flow cytometry.

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Acknowledgments

This research was supported by the Wellcome Trust (WT109965MA), NIHR Senior Fellowship (PK), NIHR Biomedical Research Centre, Oxford, NIH U19 I082360, Chinese Scholarship Council (TL). The authors wish to acknowledge the BRC Oxford GI Biobank and BRC Oxford IBD Cohort in collecting and making the samples/data available that were used in the generation of this publication.

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Authors and Affiliations

  1. Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford, Oxford, UK

    Joachim P. Hagel, Matthew Bilton, Hema Mehta, Tianqi Leng, Carl-Philipp Hackstein, Prabhjeet Phalora, Hossain D. Akther, Matthew Edmans, Christian B. Willberg & Paul Klenerman

  2. Translational Gastroenterology Unit, Nuffield Department of Medicine, University of Oxford, Oxford, UK

    Lucy C. Garner, Carl-Philipp Hackstein, Ali Amini, Nicholas M. Provine & Paul Klenerman

  3. NIHR Biomedical Research Centre, University of Oxford, Oxford, UK

    Christian B. Willberg & Paul Klenerman

Authors
  1. Joachim P. Hagel
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  2. Lucy C. Garner
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  3. Matthew Bilton
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  4. Hema Mehta
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  5. Tianqi Leng
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  6. Carl-Philipp Hackstein
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  7. Prabhjeet Phalora
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  8. Ali Amini
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  9. Hossain D. Akther
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  10. Nicholas M. Provine
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  11. Matthew Edmans
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  12. Christian B. Willberg
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  13. Paul Klenerman
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Corresponding author

Correspondence to Paul Klenerman .

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Editors and Affiliations

  1. Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden, Clinical Immunology and Transfusion Medicine, Karolinska University Hospital, Stockholm, Sweden

    Helen Kaipe

  2. Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden

    Isabelle Magalhaes

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Hagel, J.P. et al. (2020). Human MAIT Cell Activation In Vitro. In: Kaipe, H., Magalhaes, I. (eds) MAIT Cells. Methods in Molecular Biology, vol 2098. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0207-2_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0207-2_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0206-5

  • Online ISBN: 978-1-0716-0207-2

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