Peptide:MHC tetramer-based enrichment of epitope-specific T cells.

摘要: A basic necessity for researchers studying adaptive immunity with in vivo experimental models is an ability to identify T cells based on their cell antigen receptor (TCR) specificity. Many indirect methods are available which a bulk population of stimulated vitro specific and epitope-specific identified through the measurement functional response such as proliferation, cytokine production, or expression activation markers1. However, these only exhibiting one many possible functions, they not sensitive enough detect at naive precursor frequencies. popular alternative TCR transgenic adoptive transfer model, monoclonal from mouse seeded into histocompatible hosts create large that can be easily tracked use congenic marker antibody2,3. While powerful, this method suffers artifacts associated unphysiological frequency specificity single epitope4,5. Moreover, system cannot used investigate heterogeneity clones within polyclonal population. The ideal way study should involve direct detection endogenous repertoire using distinguishes solely by its binding cognate peptide:MHC (pMHC) complexes. The pMHC tetramers flow cytometry accomplishes this6, but limited high populations found following antigen-induced clonal expansion. In protocol, we describe coordinates magnetic enrichment technology enable extremely low lymphoid tissues3,7. With technique, comprehensively track entire mice all stages immune response.