小鼠粘膜相关不变性T细胞(MAIT)的鉴定和纯化

Q2 Immunology and Microbiology

Current Protocols in Immunology Pub Date : 2019-09-23 DOI:10.1002/cpim.89

Zhenjun Chen, Huimeng Wang, C. D’Souza, H. Koay, Bronwyn S. Meehan, Zhe Zhao, Troi J Pediongco, M. Shi, Tianyuan Zhu, Bingjie Wang, L. Kjer-Nielsen, S. Eckle, J. Rossjohn, D. Fairlie, D. Godfrey, R. Strugnell, J. McCluskey, A. Corbett

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引用次数: 11

摘要

本单元描述了用于研究粘膜相关不变性T (MAIT)细胞免疫的各种小鼠感染和免疫模型的效用:MAIT细胞可以从肺部(或其他组织/器官)中分离出来，然后使用MR1四聚体与一系列抗体联合使用流式细胞术进行鉴定和表征。研究了长滩军团菌或伤寒沙门氏菌感染或人工合成MAIT细胞抗原加Toll样受体激动剂免疫后，肺MAIT细胞的反应动力学、细胞因子谱和功能分化。在此过程中富集或扩增的MAIT细胞可用于进一步研究。一步一步的协议提供了MAIT细胞分类和自适应转移。然后可以对小鼠进行挑战，并对MAIT细胞进行跟踪和进一步检查。©2019 by John Wiley & Sons, Inc。

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

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.

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来源期刊

Current Protocols in Immunology Immunology and Microbiology-Immunology

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