Protocol

Antigen Processing

Volume 960 of the series Methods in Molecular Biology pp 309-326

Date:

Tracking Antigen-Specific CD8+ T Cells Using MHC Class I Multimers

  • Cécile AlanioAffiliated withImmunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d’Immunologie Humaine, Institut Pasteur
  • , Isabelle BouvierAffiliated withImmunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d’Immunologie Humaine, Institut Pasteur
  • , Hélène Jusforgues-SaklaniAffiliated withImmunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d’Immunologie Humaine, Institut Pasteur
  • , Matthew L. AlbertAffiliated withImmunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d’Immunologie Humaine, Institut Pasteur Email author 

* Final gross prices may vary according to local VAT.

Get Access

Abstract

The tracking of epitope-specific T cells is a useful approach for the study of adaptive immune responses. This protocol describes how Major Histocompatibility Complex Class I (MHC-I) multimers can be used to stain, enrich, and enumerate (rare) populations of CD8+ T cells specific for a given antigen. It provides the detailed steps for multimer labeling, magnetic enrichment, and cytometric analysis. Additionally, it provides informations for multiplexing experiments in order to achieve simultaneous detection of multiple antigenic specificities, and strategies for coupling the protocol with functional assays (e.g., intracellular cytokine staining). Future developments in cytometric systems (e.g., mass spectroscopy-based cytometry) and gene expression studies (e.g., single cell PCR) will extend these approaches and provide an unprecedented assessment of the immune repertoire.

Key words

CD8+ T cells MHC Class I multimers Antigen-specific T cells T-cell receptor