Tracking epitope-specific T cells.
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.
Department of Microbiology and Center for Immunology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
James J
JJ
Moon
H Hamlet
HH
Chu
Jason
J
Hataye
Antonio J
AJ
Pagán
Marion
M
Pepper
James B
JB
McLachlan
Traci
T
Zell
Marc K
MK
Jenkins
D000818:Animals; D001665:Binding Sites; D002452:Cell Count / Q000379:methods; D018929:Cell Culture Techniques; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D005819:Genetic Markers; D008280:Magnetics; D008285:Major Histocompatibility Complex; D051379:Mice; D008822:Mice, Transgenic; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology*
D018984:Epitopes, T-Lymphocyte; D005819:Genetic Markers; D011948:Receptors, Antigen, T-Cell