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Tracking epitope-specific T cells.
Title | Tracking epitope-specific T cells. |
Publication Type | Journal Article |
Year of Publication | 2009 |
Authors | Moon, JJ, H Chu, H, Hataye, J, Pagán, AJ, Pepper, M, McLachlan, JB, Zell, T, Jenkins, MK |
Journal | Nat Protoc |
Volume | 4 |
Issue | 4 |
Pagination | 565-81 |
Date Published | 2009 |
ISSN | 1750-2799 |
Keywords | Animals, Binding Sites, Cell Count, Cell Culture Techniques, Epitopes, T-Lymphocyte, Flow Cytometry, Genetic Markers, Magnetics, Major Histocompatibility Complex, Mice, Mice, Transgenic, Receptors, Antigen, T-Cell, T-Lymphocytes |
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. |
DOI | 10.1038/nprot.2009.9 |
Alternate Journal | Nat Protoc |
PubMed ID | 19373228 |
PubMed Central ID | PMC3517879 |
Grant List | F32 AI063793 / AI / NIAID NIH HHS / United States R01 AI039614 / AI / NIAID NIH HHS / United States R01 AI066018 / AI / NIAID NIH HHS / United States R37 AI027998 / AI / NIAID NIH HHS / United States |