Characterization and Purification of Mouse Mucosal-Associated Invariant T (MAIT) Cells

Zhenjun Chen; Huimeng Wang; Criselle D'Souza; Hui-Fern Koay; Bronwyn Meehan; Zhe Zhao; Troi Pediongco; Mai Shi; Tianyuan Zhu; Bingjie Wang; Lars Kjer-Nielsen; Sidonia B G Eckle; Jamie Rossjohn; David P Fairlie; Dale I Godfrey; Richard A Strugnell; James McCluskey; Alexandra J Corbett

doi:10.1002/cpim.89

Characterization and Purification of Mouse Mucosal-Associated Invariant T (MAIT) Cells

Curr Protoc Immunol. 2019 Dec;127(1):e89. doi: 10.1002/cpim.89.

Authors

Zhenjun Chen¹, Huimeng Wang¹, Criselle D'Souza², Hui-Fern Koay¹, Bronwyn Meehan¹, Zhe Zhao¹, Troi Pediongco¹, Mai Shi^{1

3}, Tianyuan Zhu¹, Bingjie Wang¹, Lars Kjer-Nielsen¹, Sidonia B G Eckle¹, Jamie Rossjohn^{4

5

6}, David P Fairlie^{7

8}, Dale I Godfrey^{1

9}, Richard A Strugnell¹, James McCluskey¹, Alexandra J Corbett¹

Affiliations

¹ Department of Microbiology and Immunology, The University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
² Human Immunology Translational Research Lab (HITRL), Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia.
³ School of Medicine, Tsinghua University, Beijing, China.
⁴ Infection and Immunity Program and the Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.
⁵ Australian Research Council Centre of Excellence in Advanced Molecular Imaging, Monash University, Clayton, Victoria, Australia.
⁶ Institute of Infection and Immunity, Cardiff University, School of Medicine, Heath Park, Wales, United Kingdom.
⁷ Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia.
⁸ Australian Research Council Centre of Excellence in Advanced Molecular Imaging, The University of Queensland, Brisbane, Queensland, Australia.
⁹ Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, Australia.

PMID: 31763782
DOI: 10.1002/cpim.89

Abstract

This unit describes the utility of various mouse models of infection and immunization for studying mucosal-associated invariant T (MAIT) cell immunity: MAIT cells can be isolated from the lungs (or from other tissues/organs) and then identified and characterized by flow cytometry using MR1 tetramers in combination with a range of antibodies. The response kinetics, cytokine profiles, and functional differentiation of lung MAIT cells are studied following infection with the bacterial pathogen Legionella longbeachae or Salmonella enterica Typhimurium or immunization with synthetic MAIT cell antigen plus Toll-like receptor agonist. MAIT cells enriched or expanded during the process can be used for further studies. A step-by-step protocol is provided for MAIT cell sorting and adoptive transfer. Mice can then be challenged and MAIT cells tracked and further examined. © 2019 by John Wiley & Sons, Inc.

Keywords: MR1 tetramers; flow cytometry; mouse models; mucosal-associated invariant T (MAIT) cells; respiratory infection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Female
Flow Cytometry*
Male
Mice
Mice, Inbred C57BL
Minor Histocompatibility Antigens / immunology*
Mucosal-Associated Invariant T Cells / cytology*
Mucosal-Associated Invariant T Cells / immunology*

Substances

Minor Histocompatibility Antigens