Abstract
The T-cell receptor provides T cells with specificity for antigens of particular molecular structure (the “epitope”); the T-cell pool in an individual responds to the presence of many different antigenic epitopes, but any particular T cell will respond preferentially to one defined epitope. After stimulation of a T cell by the binding of its receptor to its cognate antigen in the context of a major histocompatibility complex (MHC) molecule on an antigen-presenting cell, the T cell will begin to proliferate and synthesize cytokines. Tetramer binding and the enzyme-linked immunospot (ELISPOT) method have been used to determine what proportion of cells in the T-cell pool can bind to a defined antigenic peptide or will secrete cytokines in response to a particular antigenic stimulation. The method described here uses tracking dyes to determine what proportion of T cells will proliferate in response to stimulation. As a flow cytometric “single-cell” method, it can be combined with tetramer and cytokine staining to determine the precursor frequencies of cells in the T-cell pool able to bind tetramer, to synthesize cytokines, and to proliferate in response to antigen.
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Givan, A.L., Fisher, J.L., Waugh, M.G., Bercovici, N., Wallace, P.K. (2004). Use of Cell-Tracking Dyes to Determine Proliferation Precursor Frequencies of Antigen-Specific T Cells. In: Hawley, T.S., Hawley, R.G. (eds) Flow Cytometry Protocols. Methods in Molecular Biology™, vol 263. Humana Press. https://doi.org/10.1385/1-59259-773-4:109
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DOI: https://doi.org/10.1385/1-59259-773-4:109
Publisher Name: Humana Press
Print ISBN: 978-1-58829-234-6
Online ISBN: 978-1-59259-773-4
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