Using polyclonal TCRs specific for a single peptide:MHC complex and a sophisticated two dimensional surface approach to measure peptide/MHC:T cell interactions, Williams et al. demonstrate that peptide/MHC:TCR affinity and on rate positively correlate with T cell function and that the CD8 binding contribution to overall affinity decreases with increasing peptide/MHC:TCR affinity. Some TCRs are independent of CD8 level and would be insensitive to the natural variations in naive cell CD8 expression.
The discovery of naturally occurring T cell receptors (TCRs) that confer specific, high-affinity recognition of pathogen and cancer-associated antigens remains a major goal in cellular immunotherapies. The contribution of the CD8 co-receptor to the interaction between the TCR and peptide-bound major histocompatibility complex (pMHC) has previously been correlated with the activation and responsiveness of CD8+ T cells. However, these studies have been limited to model systems of genetically engineered hybridoma TCRs or transgenic mouse TCRs against either a single epitope or an array of altered peptide ligands. CD8 contribution in a native human antigen-specific T cell response remains elusive. Here, using Hepatitis C Virus-specific precursor CTLs spanning a large range of TCR affinities, we discovered that the functional responsiveness of any given TCR correlated with the contribution of CD8 to TCR/pMHC binding. Furthermore, we found that CD8 contribution to TCR/pMHC binding in the two-dimensional (2D) system was more accurately reflected by normalized synergy (CD8 cooperation normalized by total TCR/pMHC bonds) rather than synergy (total CD8 cooperation) alone. While synergy showed an increasing trend with TCR affinity, normalized synergy was demonstrated to decrease with the increase of TCR affinity. Critically, normalized synergy was shown to correlate with CTL functionality and peptide sensitivity, corroborating three-dimensional (3D) analysis of CD8 contribution with respect to TCR affinity. In addition, we identified TCRs that were independent of CD8 for TCR/pMHC binding. Our results resolve the current discrepancy between 2D and 3D analysis on CD8 contribution to TCR/pMHC binding, and demonstrate that naturally occurring high-affinity TCRs are more capable of CD8-independent interactions that yield greater functional responsiveness even with CD8 blocking. Taken together, our data suggest that addition of the normalized synergy parameter to our previously established TCR discovery platform using 2D TCR affinity and sequence test would allow for selection of TCRs specific to any given antigen with the desirable attributes of high TCR affinity, CD8 co-receptor independence and functional superiority. Utilizing TCRs with less CD8 contribution could be beneficial for adoptive cell transfer immunotherapies using naturally occurring or genetically engineered T cells against viral or cancer-associated antigens.
Author Info: (1) Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, United States. (2) Department of Biomedical Engineering, University of Texas at Austin, Austin,
Author Info: (1) Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, United States. (2) Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, United States. (3) McKetta Department of Chemical Engineering, University of Texas at Austin, Austin, TX, United States. (4) Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, TX, United States. (5) Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, United States. (6) Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, United States. Immunology Graduate Group, University of Pennsylvania, Philadelphia, PA, United States. (7) LIVESTRONG Cancer Institutes, Dell Medical School, University of Texas at Austin, Austin, TX, United States. (8) Center for Cell Engineering, Department of Medicine, Memorial Sloan Kettering Cancer Center (MSKCC), New York, NY, United States. Parker Institute for Cancer Immunotherapy, MSKCC, New York, NY, United States. (9) Department of Biomedical Engineering, University of Texas at Austin, Austin, TX, United States. Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, TX, United States.
