Journal
NATURE PROTOCOLS
Volume 4, Issue 4, Pages 565-581Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.9
Keywords
-
Categories
Funding
- National Institutes of Health
- American Heart Association
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R01AI039614, F32AI063793, R01AI066018, R37AI027998] Funding Source: NIH RePORTER
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available