The Impact of Altered T Cell Receptor—Peptide-Major Histocompatibility Complex Interactions on Antigen Recognition and T Cell Function

Adoptive cell transfer (ACT) using T cell receptor (TCR) gene-modified T cells is an exciting and rapidly evolving field. Numerous basic science and clinical studies have demonstrated various levels of feasibility, safety, and efficacy using TCR-engineered T cells to treat cancer and viral infections. Genomic instability of targeted diseases, however, requires effective and safe TCRs to cross-recognize mutated antigens while minimizing on- or off-target toxicities. Thus, improvements to T cell-based therapeutics mandate a broader understanding of the principles governing antigen recognition. This dissertation addresses critical biologic questions evaluating which parameters are most important in facilitating antigen recognition, and how alterations in TCR-peptide major histocompatibility complex (pMHC) interactions affect T cell function. Our model to study antigen recognition utilizes traditional and novel approaches characterizing the capability of T cells gene-modified to express an HLA-A2-restricted, cross-reactive TCR to recognize naturally occurring mutant hepatitis C virus NS3:1406-1415 epitopes. Contrary to what is generally accepted in the field, we found that TCR-pMHC affinity is not necessarily the most important factor dictating antigen recognition. Other cellular parameters, including ligand density, TCR density, and co-receptor signaling greatly influenced the recognition of altered pMHC ligands. Additionally, we found that the field’s interpretation of antigen recognition may be misguided when evaluation of T cell function is limited to a single cytokine. Functional phenotypes by seven-parameter flow cytometry revealed that T cell functional profiles are more complex than were previously believed, and evaluation of a single functional phenotype did not accurately reflect the functional behavior of a T cell culture. Combining functional studies with structural analysis of the TCR-pMHC interface helps bring clarity to these unexpected results. Together, our data suggest that the field is oversimplifying T cell function and establish a new working model highlighting a previously unappreciated and complex relationship between kinetic, cellular, and structural parameters governing antigen recognition. This enhanced understanding will not only help steer rational, structure-guided design of TCRs to generate better functioning T cells for ACT, but will also impact the way in which we design, evaluate, and implement novel immunotherapies

Physical and biological variables affecting seabird distributions during the upwelling season of the northern California Current

Author Posting. © The Authors, 2004. This is the author's version of the work. It is posted here by permission of Elsevier B. V. for personal use, not for redistribution. The definitive version was published in Deep Sea Research Part II: Topical Studies in Oceanography 52 (2005): 123-143, doi:10.1016/j.dsr2.2004.08.016.As a part of the GLOBEC-Northeast Pacific project, we investigated variation in the abundance of marine birds in the context of biological and physical habitat conditions in the northern portion of the California Current System (CCS) during cruises during the upwelling season 2000. Continuous surveys of seabirds were conducted simultaneously in June (onset of upwelling) and August (mature phase of upwelling) with ocean properties quantified using a towed, undulating vehicle and a multi-frequency bioacoustic instrument (38-420 kHz). Twelve species of seabirds contributed 99% of the total community density and biomass. Species composition and densities were similar to those recorded elsewhere in the CCS during earlier studies of the upwelling season. At a scale of 2-4 km, physical and biological oceanographic variables explained an average of 25% of the variation in the distributions and abundance of the 12 species. The most important explanatory variables (among 14 initially included in each multiple regression model) were distance to upwelling-derived frontal features (center and edge of coastal jet, and an abrupt, inshore temperature gradient), sea-surface salinity, acoustic backscatter representing various sizes of prey (smaller seabird species were associated with smaller prey and the reverse for larger seabird species), and chlorophyll concentration. We discuss the importance of these variables in the context of what factors may be that seabirds use to find food. The high seabird density in the Heceta Bank and Cape Blanco areas indicate them to be refuges contrasting the low seabird densities currently found in most other parts of the CCS, following decline during the recent warm regime of the Pacific Decadal Oscillation.Support from National Science Foundation Grant OCE-0001035, National Oceanic and Atmospheric Administration (NOAA)/Woods Hole Oceanographic Institution-CICOR Grant NA17RJ1223 is gratefully acknowledged

How an alloreactive T-cell receptor achieves peptide and MHC specificity

T-cell receptor (TCR) allorecognition is often presumed to be relatively nonspecific, attributable to either a TCR focus on exposed major histocompatibility complex (MHC) polymorphisms or the degenerate recognition of allopeptides. However, paradoxically, alloreactivity can proceed with high peptide and MHC specificity. Although the underlying mechanisms remain unclear, the existence of highly specific alloreactive TCRs has led to their use as immunotherapeutics that can circumvent central tolerance and limit graft-versus-host disease. Here, we show how an alloreactive TCR achieves peptide and MHC specificity. The HCV1406 TCR was cloned from T cells that expanded when a hepatitis C virus (HCV)-infected HLA-A2− individual received an HLA-A2+ liver allograft. HCV1406 was subsequently shown to recognize the HCV nonstructural protein 3 (NS3):1406– 1415 epitope with high specificity when presented by HLA-A2. We show that NS3/HLA-A2 recognition by the HCV1406 TCR is critically dependent on features unique to both the allo-MHC and the NS3 epitope. We also find cooperativity between structural mimicry and a crucial peptide “hot spot” and demonstrate its role, along with the MHC, in directing the specificity of allorecognition. Our results help explain the paradox of specificity in alloreactive TCRs and have implications for their use in immunotherapy and related efforts to manipulate TCR recognition, as well as alloreactivity in general

How an alloreactive T-cell receptor achieves peptide and MHC specificity

T-cell receptor (TCR) allorecognition is often presumed to be relatively nonspecific, attributable to either a TCR focus on exposed major histocompatibility complex (MHC) polymorphisms or the degenerate recognition of allopeptides. However, paradoxically, alloreactivity can proceed with high peptide and MHC specificity. Although the underlying mechanisms remain unclear, the existence of highly specific alloreactive TCRs has led to their use as immunotherapeutics that can circumvent central tolerance and limit graft-versus-host disease. Here, we show how an alloreactive TCR achieves peptide and MHC specificity. The HCV1406 TCR was cloned from T cells that expanded when a hepatitis C virus (HCV)-infected HLA-A2− individual received an HLA-A2+ liver allograft. HCV1406 was subsequently shown to recognize the HCV nonstructural protein 3 (NS3):1406– 1415 epitope with high specificity when presented by HLA-A2. We show that NS3/HLA-A2 recognition by the HCV1406 TCR is critically dependent on features unique to both the allo-MHC and the NS3 epitope. We also find cooperativity between structural mimicry and a crucial peptide “hot spot” and demonstrate its role, along with the MHC, in directing the specificity of allorecognition. Our results help explain the paradox of specificity in alloreactive TCRs and have implications for their use in immunotherapy and related efforts to manipulate TCR recognition, as well as alloreactivity in general