Tracking epitope-specific T cells

JJ Moon, HH Chu, J Hataye, AJ Pagán, M Pepper… - Nature protocols, 2009 - nature.com
JJ Moon, HH Chu, J Hataye, AJ Pagán, M Pepper, JB McLachlan, T Zell, MK Jenkins
Nature protocols, 2009nature.com
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 …
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.
nature.com