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T Cell Response in the Lung Following Influenza Virus Infection

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1591)

Abstract

Methods that enable the identification of virus-specific CD4 and CD8 T cells are key to our understanding of how the adaptive immune response controls viral infection. Here we describe two distinct methods to evaluate the T cell response to influenza A virus (IAV). The number and phenotype of T cells that respond to natural IAV epitopes can be assessed by flow cytometry using MHC class I and class II tetramers. Using this system, IAV-specific T cells can be tracked in various organs within the same animal, or, in different cohorts, the response can be evaluated at several time points following infection. While providing clear quantitative data, flow cytometry cannot provide any information about T cell location within the lung or interactions between responding T cells and other cell types. Here we also describe a method to examine activated CD4 T cells in the lungs of living animals using multiphoton intravital microscopy, thus providing real-time analysis of T cell behavior during an infection.

Keywords

T cells MHC tetramers Lung Influenza A virus Intravital imaging Memory Infection 
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Notes

Acknowledgments

We are grateful to Prof. Matthew Krummel, University of California, San Francisco, and his laboratory for sharing the methodology of the intravital lung imaging technique and to Prof. James Brewer and Dr. Ross McQueenie for their assistance in setting up this method at Glasgow University. We acknowledge the NIH Tetramer Core Facility (contract HHSN272201300006C) for provision of MHC tetramers.

Dr. MacLeod is an Arthritis Research UK Fellow (Grant ID: 19905).

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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Centre for Immunobiology, Institute of Infection, Immunity and InflammationThe University of GlasgowGlasgowUK
  2. 2.Centre for Immunobiology, Institute of Infection, Immunity and InflammationThe University of GlasgowGlasgowUK

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