Identification of Functional HLA-A*01:01–Restricted Epstein-Barr Latent Membrane Protein 2–Specific T-Cell Receptors
| Autor: | Lieve E. van der Maarel, J.H. Frederik Falkenburg, Rob C. M. de Jong, Ilse Gille, Inge Jedema, Wesley Huisman, Derk Amsen, Lois Hageman, Laura T. Morton, Mirjam H.M. Heemskerk |
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| Přispěvatelé: | Graduate School, Medical Biology, Landsteiner Laboratory, AII - Inflammatory diseases |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Epstein-Barr Virus Infections Herpesvirus 4 Human Adoptive cell transfer Receptors Antigen T-Cell lymphoma chemical and pharmacologic phenomena Human leukocyte antigen Biology medicine.disease_cause virus-specific T cells Viral Matrix Proteins EBV-associated malignancies Interferon-gamma 03 medical and health sciences 0302 clinical medicine hemic and lymphatic diseases otorhinolaryngologic diseases medicine Humans Epstein-Barr virus Immunology and Allergy Secretion Receptor Cytotoxicity HLA-A Antigens nasopharyngeal carcinoma T-cell receptor Epstein–Barr virus Molecular biology HLA-A 030104 developmental biology Infectious Diseases TCR-gene transfer 030220 oncology & carcinogenesis |
| Zdroj: | Journal of infectious diseases, 226(5), 833-842. Oxford University Press The Journal of Infectious Diseases, 226(5), 833-842. OXFORD UNIV PRESS INC |
| ISSN: | 1537-6613 0022-1899 |
| DOI: | 10.1093/infdis/jiaa512 |
| Popis: | Background Adoptive transfer of genetically engineered T cells expressing antigen-specific T-cell receptors (TCRs) is an appealing therapeutic approach for Epstein-Barr virus (EBV)–associated malignancies of latency type II/III that express EBV antigens (LMP1/2). Patients who are HLA-A*01:01 positive could benefit from such products, since no T cells recognizing any EBV-derived peptide in this common HLA allele have been found thus far. Methods HLA-A*01:01–restricted EBV-LMP2–specific T cells were isolated using peptide major histocompatibility complex (pMHC) tetramers. Functionality was assessed by production of interferon gamma (IFN-γ) and cytotoxicity when stimulated with EBV-LMP2–expressing cell lines. Functionality of primary T cells transduced with HLA-A*01:01–restricted EBV-LMP2–specific TCRs was optimized by knocking out the endogenous TCRs of primary T cells (∆TCR) using CRISPR-Cas9 technology. Results EBV-LMP2–specific T cells were successfully isolated and their TCRs were characterized. TCR gene transfer in primary T cells resulted in specific pMHC tetramer binding and reactivity against EBV-LMP2–expressing cell lines. The mean fluorescence intensity of pMHC-tetramer binding was increased 1.5–2 fold when the endogenous TCRs of CD8+ T cells was knocked out. CD8+/∆TCR T cells modified to express EBV-LMP2–specific TCRs showed IFN-γ secretion and cytotoxicity toward EBV-LMP2–expressing malignant cell lines. Conclusions We isolated the first functional HLA-A*01:01–restricted EBV-LMP2–specific T-cell populations and TCRs, which can potentially be used in future TCR gene therapy to treat EBV-associated latency type II/III malignancies. |
| Databáze: | OpenAIRE |
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