title: TCRs genetically linked to CD28 and CD3ε do not mispair with endogenous TCR chains and mediate enhanced T cell persistence and anti-melanoma activity.
abstract: Adoptive transfer of T cells that are gene engineered to express a defined TCR represents a feasible and promising therapy for patients with tumors. However, TCR gene therapy is hindered by the transient presence and effectiveness of transferred T cells, which are anticipated to be improved by adequate T cell costimulation. In this article, we report the identification and characterization of a novel two-chain TCR linked to CD28 and CD3ε (i.e., TCR:28ε). This modified TCR demonstrates enhanced binding of peptide-MHC and mediates enhanced T cell function following stimulation with peptide compared with wild-type TCR. Surface expression of TCR:28ε depends on the transmembrane domain of CD28, whereas T cell functions depend on the intracellular domains of both CD28 and CD3ε, with IL-2 production showing dependency on CD28:LCK binding. TCR:28ε, but not wild-type TCR, induces detectable immune synapses in primary human T cells, and such immune synapses show significantly enhanced accumulation of TCR transgenes and markers of early TCR signaling, such as phosphorylated LCK and ERK. Importantly, TCR:28ε does not show signs of off-target recognition, as evidenced by lack of TCR mispairing, as well as preserved specificity. Notably, when testing TCR:28ε in immune-competent mice, we observed a drastic increase in T cell survival, which was accompanied by regression of large melanomas with limited recurrence. Our data argue that TCR transgenes that contain CD28, and, thereby, may provide T cell costimulation in an immune-suppressive environment, represent candidate receptors to treat patients with tumors.
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Coen
authors.initials: C
authors.lastname: Govers
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Sebestyén
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, 4032 Debrecen, Hungary; and.
authors.forename: János
authors.initials: J
authors.lastname: Roszik
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Mandy
authors.initials: M
authors.lastname: van Brakel
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Cor
authors.initials: C
authors.lastname: Berrevoets
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, 4032 Debrecen, Hungary; and.
authors.forename: Árpád
authors.initials: Á
authors.lastname: Szöőr
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Konstantina
authors.initials: K
authors.lastname: Panoutsopoulou
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Marieke
authors.initials: M
authors.lastname: Broertjes
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands;
authors.forename: Tan
authors.initials: T
authors.lastname: Van
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, 4032 Debrecen, Hungary; and Cell Biology and Signaling Research Group of the Hungarian Academy of Sciences, Medical and Health Science Center, University of Debrecen, 4032 Debrecen, Hungary.
authors.forename: György
authors.initials: G
authors.lastname: Vereb
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, 4032 Debrecen, Hungary; and Cell Biology and Signaling Research Group of the Hungarian Academy of Sciences, Medical and Health Science Center, University of Debrecen, 4032 Debrecen, Hungary.
authors.forename: János
authors.initials: J
authors.lastname: Szöllősi
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center Cancer Institute, 3015 CN Rotterdam, the Netherlands; j.debets@erasmusmc.nl.
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
mesh_terms: D000818:Animals; D018106:CD28 Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017252:CD3 Complex / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D054992:Immunological Synapses; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000235:genetics / Q000276:immunology; D008545:Melanoma / Q000235:genetics / Q000276:immunology / Q000401:mortality / Q000628:therapy*; D051379:Mice; D009364:Neoplasm Recurrence, Local / Q000517:prevention & control; D009368:Neoplasm Transplantation; D011485:Protein Binding; D015202:Protein Engineering; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D012878:Skin Neoplasms / Q000235:genetics / Q000276:immunology / Q000401:mortality / Q000628:therapy*; D016019:Survival Analysis; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D047368:Tumor Burden
chemical_list: D018106:CD28 Antigens; D017252:CD3 Complex; C115335:CD3E protein, human; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D048049:Extracellular Signal-Regulated MAP Kinases