Tracking antigen-specific CD8⁺ T cells using MHC class I multimers

Immunobiologie des cellules dendritiques, Unité Mixte Pasteur/Inserm U818, Institut Pasteur and Centre d'Immunologie Humaine, Institut Pasteur, Paris, France.
Methods in molecular biology (Clifton, N.J.) (Impact Factor: 1.29). 01/2013; 960:309-326. DOI: 10.1007/978-1-62703-218-6_23
Source: PubMed


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.

Download full-text


Available from: Cécile Alanio, Jul 08, 2014
28 Reads
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Multimer monitoring has become a standard technique for detection of antigen-specific T cells. The term "multimer" refers to a group of reagents based on the multimerisation of molecules in order to raise avidity and thus stabilize binding to their ligand. Multimers for detection of antigen-specific T cell responses are based on major histocompatibility complex class I peptide complexes. Multimer staining enables fast and direct visualization of antigen-specific T cells, thus it is widely applied to assess antiviral immunity, e.g. monitor patients in vaccination trials or confirm purity of cell products for adoptive transfer. Assessment of T cell immunity against persistent pathogens like e.g. cytomegalovirus (CMV) is of major importance in immunosuppressed patients. Recent advancements of multimers facilitate reversible labeling and allow isolation of epitope-specific T cells for adoptive transfer. Here, we give an overview on the different multimers and their applications, with an emphasis on CMV-specific T cell responses.
    Diagnostic microbiology and infectious disease 01/2013; 78(3). DOI:10.1016/j.diagmicrobio.2013.11.007 · 2.46 Impact Factor