title: Broadening Specificity and Enhancing Cytotoxicity of Adoptive T Cells for Nasopharyngeal Carcinoma Immunotherapy.
abstract: Although promising, clinical responses to adoptive immunotherapy for nasopharyngeal carcinoma (NPC) are still limited by the restricted number of Epstein-Barr virus (EBV) antigens that can be targeted and their poor immunogenicity. Our previous work indicated that the immunogenic features of the NPC-associated viral antigen BARF1 may be exploited for immunotherapeutic purposes. Nevertheless, T-cell lines obtained with current protocols include only negligible numbers of BARF1-specific cytotoxic T lymphocytes, pointing to the need to enrich these effectors in BARF1 specificities. Considering that in B lymphocytes BARF1 is mainly a lytic EBV antigen, we tested different EBV lytic-cycle inducers (TPA/butyric acid, doxorubicin, and cisplatin) used at suboptimal concentrations for their ability to upregulate BARF1 expression in lymphoblastoid B-cell lines (LCL), the commonly used antigen-presenting cells, without compromising their survival. The LCLs treated with doxorubicin (DX-LCL) can reproducibly and efficiently generate EBV-specific effectors enriched in BARF1 specificities from both healthy donors and NPC patients. These DX-LCLs also had more pronounced immunogenic properties, including HLA class I upregulation and expression of immunogenic cell death markers, such as enhanced calreticulin exposure and HMGB1 release. In particular, doxorubicin triggers an HMGB1 autocrine/paracrine loop with its receptor, TLR4, which is also upregulated in DX-LCLs and is responsible for NF-κB activation and a delayed apoptosis that allows a prolonged stimulation of EBV-specific T-cell precursors. This protocol may thus constitute a valid alternative to the use of engineered LCLs to generate EBV-specific T-cell lines for adoptive immunotherapy, being relatively simple, easily upgradable to Good Manufacturing Practice standards, and therefore more broadly applicable. Cancer Immunol Res; 4(5); 431-40. ©2016 AACR.
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Damiana Antonia
authors.initials: DA
authors.lastname: Faè
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Debora
authors.initials: D
authors.lastname: Martorelli
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Katy
authors.initials: K
authors.lastname: Mastorci
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Muraro
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Jessica
authors.initials: J
authors.lastname: Dal Col
authors.affiliation: Division of Radiotherapy, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Franchin
authors.affiliation: Otolaryngology, Azienda Ospedaliera S. Maria degli Angeli, Pordenone, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Barzan
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Comaro
authors.affiliation: Medical Oncology Division A, CRO-IRCCS, National Cancer Institute, Aviano, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Vaccher
authors.affiliation: Istituto Oncologico Veneto, IOV-IRCCS, Padua, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rosato
authors.affiliation: Cancer Bio-Immunotherapy Unit, CRO-IRCCS, National Cancer Institute, Aviano, Italy. r.dolcetti@uq.edu.au.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Dolcetti
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000938:Antigen-Presenting Cells / Q000276:immunology; D002277:Carcinoma; D002454:Cell Differentiation / Q000276:immunology; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004317:Doxorubicin / Q000494:pharmacology; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000903:Antibiotics, Antineoplastic; C086495:BARF1 protein, Human herpesvirus 4; D014764:Viral Proteins; D004317:Doxorubicin