Identification of Functional HLA-A*01:01–Restricted Epstein-Barr Latent Membrane Protein 2–Specific T-Cell Receptors
- Resource Type
- Authors
- 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
- Source
- Journal of infectious diseases, 226(5), 833-842. Oxford University Press
The Journal of Infectious Diseases, 226(5), 833-842. OXFORD UNIV PRESS INC
- Subject
- 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
- Language
- ISSN
- 1537-6613
0022-1899
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.