Human MAIT cell activation in vitro

Book section

Item description: Chapter 7 in book 'MAIT Cells' (2020), ed H Kaipe and I Magalhaes. Methods in Molecular Biology, volume 2098.
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.

Authors: Hagel, JP; Garner, LC; Bilton, M; Mehta, H; Leng, T

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Humana Press
• Volume: 2098
• Pages: 97-124
• Series: Methods in Molecular Biology
• Place of publication: New York, USA
• Publication date: 2019-12-03
• Event title: MAIT Cells
• DOI: 10.1007/978-1-0716-0207-2_7
• EISSN: 1940-6029
• ISSN: 1064-3745
• Pmid: 31792818
• EISBN: 978-1-0716-0207-2
• ISBN: 978-1-0716-0206-5
• Language: English
• Keywords: beads; cytokines; interleukin-18; 5-OP-RU; CD28; CD3; Vα7.2; invariant T cell receptor; MAIT cells; FFR; innate; SBTMR; toll-like receptor; MR1; interleukin-12; E. coli; plate-bound