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Simultaneous Analysis of HCV-Specific CD4+ and CD8+ T Cells by Multicolor Flow Cytometry

  • David Wolski
  • Georg M. LauerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1911)

Abstract

CD4 T cell responses are key to effective control of HCV infection. However, their precise mechanisms of action and the molecular programs leading to effective T help versus CD4 T cell failure remain elusive. Direct visualization of HCV-specific CD4 T cells using HLA class II tetramers holds the promise to better define the function and phenotype of these cells and to isolate them for further molecular analysis. Here we describe how to utilize peptide-MHC (pMHC) class II tetramers in multicolor flow cytometry to define the expression of molecules on the surface and within HCV-specific CD4 T cells, how to analyze HCV-specific CD4 and CD8 T cells in the same tube, and how to sort live HCV-specific CD4 T cells as single cells or T cell populations for further analysis by RNAseq or other methods.

Key words

HCV-specific T cells CD4 CD8 Cell sorting Flow cytometry Tetramer Pentamer Peptide-MHC Intracellular staining Transcription factor 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Gastrointestinal Unit and Liver Center, Massachusetts General Hospital andHarvard Medical SchoolBostonUSA

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