title: Monitoring the Dynamics of T Cell Clonal Diversity Using Recombinant Peptide:MHC Technology.
abstract: The capacity to probe antigen specific T cells within the polyclonal repertoire has been revolutionized by the advent of recombinant peptide:MHC (pMHC) technology. Monomers and multimers of pMHC molecules can enrich for and identify antigen specific T cells to elucidate the contributions of T cell frequency, localization, and T cell receptor (TCR) affinity during immune responses. Two-dimensional (2D) measurements of TCR-pMHC interactions are at the forefront of this field because the biological topography is replicated such that TCR and pMHC are membrane anchored on opposing cells, allowing for biologically pertinent measures of TCR antigen specificity and diversity. 2D measurements of TCR-pMHC kinetics have also demonstrated increased fidelity compared to three-dimensional surface plasmon resonance data and are capable of detecting T cell affinities that are below the detection level of most pMHC multimers. Importantly, 2D techniques provide a platform to evaluate T cell affinity and antigen specificity against multiple protein epitopes within the polyclonal repertoire directly ex vivo from sites of ongoing immune responses. This review will discuss how antigen specific pMHC molecules, with a focus on 2D technologies, can be used as effective tools to evaluate the range of TCR affinities that comprise an immune response and more importantly how the breadth of affinities determine functional outcome against a given exposure to antigen.
authors.affiliation: Department of Microbiology and Immunology, Emory University, Atlanta, GA , USA.
authors.forename: J Lori
authors.initials: JL
authors.lastname: Blanchfield
authors.forename: Shayla K
authors.initials: SK
authors.lastname: Shorter
authors.forename: Brian D
authors.initials: BD
authors.lastname: Evavold
keywords: 2D assays; T cell activation; T cell affinity; kinetics; recombinant pMHC