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Human MAIT Cell Activation In Vitro.

Joachim P Hagel1, Lucy C Garner2, Matthew Bilton1

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

Methods in Molecular Biology (Clifton, N.J.)
|December 4, 2019
PubMed
Summary
This summary is machine-generated.

Mucosal-associated invariant T (MAIT) cells can be activated through T cell receptor (TCR)-dependent or -independent pathways. This study details methods for activating MAIT cells and analyzing them via flow cytometry.

Keywords:
5-OP-RUBeadsCD28CD3CytokinesE. coliInnateInterleukin-12Interleukin-18Invariant T cell receptorMAIT cellsMR1Plate-boundToll-like receptorVα7.2

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Area of Science:

  • Immunology
  • Cellular Biology

Background:

  • Mucosal-associated invariant T (MAIT) cells are a key innate-like T cell population found in human mucosal tissues and the liver.
  • MAIT cells possess a semi-invariant T cell receptor (TCR) that recognizes microbial riboflavin metabolites presented by the MHC Class I-like molecule MR1.

Purpose of the Study:

  • To describe various methods for activating MAIT cells, encompassing both TCR-dependent and TCR-independent pathways.
  • To provide a protocol for analyzing activated MAIT cells using flow cytometry.

Main Methods:

  • TCR-dependent activation involved stimulation with anti-CD3/anti-CD28 antibodies, 5-OP-RU, or PFA-fixed E. coli with antigen-presenting cells (APCs).
  • TCR-independent activation utilized direct stimulation with IL-12 and IL-18, or indirect stimulation with TLR agonists or influenza A virus in the presence of APCs.
  • Flow cytometry was employed to analyze the activation status of MAIT cells.

Main Results:

  • Successful activation of MAIT cells was achieved through both TCR-dependent and TCR-independent routes.
  • The study outlines specific protocols for each activation method, detailing the reagents and conditions required.
  • Analysis of activated MAIT cells via flow cytometry is demonstrated.

Conclusions:

  • MAIT cells exhibit versatile activation mechanisms, responding to both TCR engagement and cytokine signaling.
  • The described methods provide a comprehensive toolkit for researchers studying MAIT cell function and activation.
  • This work facilitates further investigation into the role of MAIT cells in immunity and disease.