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Nat Protoc. 2009;4(4):565-81. doi: 10.1038/nprot.2009.9.

Tracking epitope-specific T cells.

Author information

1
Department of Microbiology and Center for Immunology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.

Abstract

The tracking of antigen-specific T cells in vivo is a useful approach for the study of the adaptive immune response. This protocol describes how populations of T cells specific for a given peptide-major histocompatibility complex (pMHC) epitope can be tracked based solely on T-cell receptor (TCR) specificity as opposed to other indirect methods based on function. The methodology involves the adoptive transfer of TCR transgenic T cells with defined epitope specificity into histocompatible mice and the subsequent detection of these cells through the use of congenic or clonotypic markers. Alternatively, endogenous epitope-specific T cells can be tracked directly through the use of pMHC tetramers. Using magnetic bead-based enrichment and advanced multiparameter flow cytometry, populations as small as five epitope-specific T cells can be detected from the peripheral lymphoid organs of a mouse. The adoptive transfer procedure can be completed within 3 h, whereas analysis of epitope-specific cells from mice can be completed within 6 h.

PMID:
19373228
PMCID:
PMC3517879
DOI:
10.1038/nprot.2009.9
[Indexed for MEDLINE]
Free PMC Article

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