Human MAIT Cell Activation In Vitro
| Autor: | Nicholas M. Provine, Joachim Hagel, Christian B. Willberg, Carl-Philipp Hackstein, Lucy C. Garner, Paul Klenerman, Matthew Bilton, Ali Amini, Tianqi Leng, Prabhjeet Phalora, Matthew Edmans, Hema Mehta, Hossain Delowar Akther |
|---|---|
| Přispěvatelé: | Kaipe, H, Magalhaes, I |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
CD3 Receptors Antigen T-Cell Antigen-Presenting Cells Lymphocyte Activation Mucosal-Associated Invariant T Cells Flow cytometry Cell Line Immunophenotyping 03 medical and health sciences 0302 clinical medicine T-Lymphocyte Subsets MHC class I medicine Escherichia coli Humans Cells Cultured Toll-like receptor medicine.diagnostic_test biology Staining and Labeling Chemistry T-cell receptor Toll-Like Receptors CD28 Flow Cytometry Cell biology 030104 developmental biology Viruses Interleukin 12 biology.protein Cytokines Cell activation Biomarkers 030215 immunology |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071602065 |
| ISSN: | 1940-6029 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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