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Tracking epitope-specific T cells
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DOI：10.1038/nprot.2009.9.png)
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摘要
En 中文
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.
Keyword:
CLASS-II TETRAMERS
EX-VIVO ANALYSIS
IN-VIVO
PRECURSOR FREQUENCY
FUNCTIONAL AVIDITY
PEPTIDE TETRAMERS
ANTIGEN-RECEPTOR
CLONAL EXPANSION
PERIPHERAL-BLOOD
IMMUNE-RESPONSE
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IF:
16
论文数:
4.0K
被引数:
5.6W
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