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Tracking epitope-specific T cells

Nature Protocols volume 4, pages 565–581 (2009)Cite this article

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.

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Figure 1: Graphical representation of in vivo T-cell-tracking strategies.
Figure 2: Analysis of TCR transgenic T cells before adoptive transfer.
Figure 3: Analysis of donor TCR transgenic T cells from a high-frequency adoptive transfer.
Figure 4: Analysis of donor TCR transgenic T cells from a low-frequency adoptive transfer.
Figure 5: Analysis of endogenous epitope-specific T cells.

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Acknowledgements

This work was supported by the National Institutes of Health (J.J.M., J.H., A.J.P., M.P., J.B.M. and M.K.J.) and the American Heart Association (H.H.C.).

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  1. Jason Hataye

    Present address: Present address: Department of Medicine, David Geffen School of Medicine, University of California-Los Angeles, Los Angeles, California 90095, USA.,

  2. Traci Zell

    Present address: Present address: eBioscience, 10255 Science Center Drive, San Diego, California 92121, USA.,

Authors and Affiliations

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

    James J Moon, H Hamlet Chu, Jason Hataye, Antonio J Pagán, Marion Pepper, James B McLachlan & Marc K Jenkins

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Moon, J., Chu, H., Hataye, J. et al. Tracking epitope-specific T cells. Nat Protoc 4, 565–581 (2009). https://doi.org/10.1038/nprot.2009.9

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