期刊
NATURE PROTOCOLS
卷 4, 期 4, 页码 565-581出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.9
关键词
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资金
- National Institutes of Health
- American Heart Association
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R01AI039614, F32AI063793, R01AI066018, R37AI027998] Funding Source: NIH RePORTER
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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