MAIT Cells Methods and Protocols

This volume focuses on various methods used by researchers to study mucosal-associated invariant T (MAIT) cells in the aspects described below. The chapters are organized into Four Parts: part One looks at the function and importance of MAIT cells in health and disease. It covers methods to isolate and characterize MAIT cells from human tissues including liver, colon tumors, placenta and decidua, and endometrium. Part Two discusses MR1 independent stimulation, and looks at the activation of MAIT cells by different stimulatory agents. Part Three describes various methods to develop tools to analyze, monitor, and generate MAIT cells in vitro, including the production of MR1-tetramers and how they can be loaded with bacterial antigens and used for the detection of MAIT cells. The last part covers the use of murine models to study MAIT cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and comprehensive, MAIT Cells: Methods and Protocols is a valuable resource for anyone interested in learning more about this developing field.

Chapters 9 and 17 are open access under a CC BY 4.0 license.

Preface…
Table of Contents… 
Contributing Authors…

Part I MAIT Cells in Human Tissues and Blood 

1. MAIT Cells in Health and Disease
Isabelle Magalhaes, Martin Solders, and Helen Kaipe

2. Isolation and Characterization of MAIT Cells from Human Tissue Biopsies
Daniela Di Blasi, Alessandro Vacchini, Gennaro De Libero, and Lucia Mori

3. Isolation and Characterization of MAIT Cells from Tumor Tissues
William Rodin, Patrik Sundström, and Marianne Quiding Järbrink

4. Isolation of Immune Cells from Placental Tissues and Phenotypic and Functional Analysis of MAIT Cells
Martin Solders, Laia Gorchs, and Helen Kaipe

5. Methods for High-Dimensional Flow Cytometry Analysis of Human MAIT Cells in Tissues and Peripheral Blood
Benedikt Strunz, Christine L. Zimmer, Jonna Bister, Martin A. Ivarsson, and Niklas K. Björkström

6. In Situ Detection of MAIT Cells and MR1 Expressing Cells in Tissue Biopsies Utilizing Immunohistochemistry
Anna Gibbs, Michal J. Sobkowiak, Johan K. Sandberg, and Annelie Tjernlund

Part II Functional Analysis of MAIT Cells

7. Human MAIT Cell Activation In Vitro
Joachim P. Hagel, Lucy C. Garner, Matthew Bilton, Hema Mehta, Tianqi Leng, Carl-Philipp Hackstein, Prabhjeet Phalora, Ali Amini, Hossain D. Akther, Nicholas M Provine, Matthew Edmans, Christian B. Willberg, and Paul Klenerman  

8. Interactions between MAIT Cells and Dendritic Cells
Mariolina Salio and Vincenzo Cerundolo

9. Influenza A Virus-Infected Lung Epithelial Cell Co-Culture with Human Peripheral Blood Mononuclear Cells
Liyen Loh, Marios Koutsakos, Katherine Kedzierska, and Timothy S.C. Hinks

10. Quantification of Human MAIT Cell-Mediated Cellular Cytotoxicity and Antimicrobial Activity
Wan Rong Sia, Caroline Boulouis, Muhammad Yaaseen Gulam, Andrea Lay Hoon Kwa, Johan K. Sandberg, and Edwin Leeansyah

11. MAIT Cell Activation by Fungal Pathogens
Susanne Jahreis, Sarah Boettcher, and Marie von Lilienfeld-Toal

12. A Flow Chamber Assay for Studying MAIT Cell Trafficking
Farhat Parween, Hongwei H. Zhang, and Joshua M. Farber

Part III Monitoring and In Vitro Amplification of MAIT Cells 

13. Production of MR1 Tetramers Loaded with Microbial Ligands
Nicole A. Ladd, Cara A. Froyd, and Erin J. Adams

14. Immune Monitoring of Human Mucosal Associated Invariant T Cells by Quantitative Proteomics 
Isabel Bernal, Marco van Ham, and Lothar Jänsch

15. Generation of MR1-Restricted T Cell Clones by Limiting Dilution Cloning of MR1 Tetramer+ Cells
Meghan Cansler, Megan Null, Erin Meermeier, Gwendolyn Swarbrick, David Lewinsohn, and Melanie Harriff

16. Reprogramming of MAIT Cells to Pluripotency and Redifferentiation
Hiroshi Wakao

Part IV MAIT Cells in Murine Models 

17. Study of MAIT Cell Activation in Viral Infections In Vivo
Timothy S.C. Hinks, Bonnie van Wilgenburg, Huimeng Wang, Liyen Loh, Marios Koutsakos, Katherine Kedzierska, Alexandra J. Corbett, and Zhenjun Chen 

18. MAIT Cells in Type 1 Diabetes Mouse Models
Isabelle Nel, Lucie Beaudoin, and Agnès Lehuen

19. Magnetic-Bead Based Enrichment of Murine MAIT Cells
Samantha J. Winter and Andreas Krueger

Subject Index List…