Abstract
MAIT cells are abundant, highly evolutionarily conserved innate-like lymphocytes expressing a semi-invariant T cell receptor (TCR), which recognizes microbially derived small intermediate molecules from the riboflavin biosynthetic pathway. However, in addition to their TCR-mediated functions they can also be activated in a TCR-independent manner via cytokines including IL-12, -15, -18, and type I interferon. Emerging data suggest that they are expanded and activated by a range of viral infections, and significantly that they can contribute to a protective anti-viral response. Here we describe methods used to investigate these anti-viral functions in vivo in murine models. To overcome the technical challenge that MAIT cells are rare in specific pathogen-free laboratory mice, we describe how pulmonary MAIT cells can be expanded using intranasal bacterial infection or a combination of synthetic MAIT cell antigen and TLR agonists. We also describe protocols for adoptive transfer of MAIT cells, methods for lung homogenization for plaque assays, and surface and intracellular cytokine staining to determine MAIT cell activation.
Highlights
MAIT cells are relatively recently described innate-like lymphocytes, with similarities to the invariant natural killer T and γδ T cell subsets [1–4]
To overcome the technical challenge that MAIT cells are rare in specific pathogen-free laboratory mice, we describe (1) how pulmonary MAIT cells can be expanded using intranasal (i.n.) bacterial infection or a combination of synthetic MAIT cell antigen and TLR agonists as well as protocols for (2) adoptive transfer of MAIT cells, (3) viral preparation and infection of mice, (4) lung homogenization, (5) surface and intracellular cytokine staining to determine MAIT cell activation, and (6) plaque assays
From a 500 mL bottle of phosphate buffered saline (PBS), add 40 mL to a 50 mL falcon containing 2.5 g bovine serum albumin (BSA) powder, vortex hard, filter sterilize back into PBS bottle using a syringe through a 0.22-μm filter
Summary
MAIT cells are relatively recently described innate-like lymphocytes, with similarities to the invariant natural killer T (iNKT) and γδ T cell subsets [1–4]. MAIT cells express a semi-invariant T cell receptor (TCR), which recognizes microbially derived small molecule intermediates from the riboflavin biosynthetic pathway [1, 4, 8, 9] These molecular intermediates exist only in microbes but not in mammals, and constitute a signature of microbial infection. Emerging data suggest that they are expanded and activated by a range of human viral infections including dengue, hepatitis C, and influenza virus [11, 13]. It was not clear from observational human studies whether this would lead to enhanced immune protection, or, contribute to immunopathology. To overcome the technical challenge that MAIT cells are rare in specific pathogen-free laboratory mice, we describe (1) how pulmonary MAIT cells can be expanded using intranasal (i.n.) bacterial infection or a combination of synthetic MAIT cell antigen and TLR agonists as well as protocols for (2) adoptive transfer of MAIT cells, (3) viral preparation and infection of mice, (4) lung homogenization, (5) surface and intracellular cytokine staining to determine MAIT cell activation, and (6) plaque assays
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