title: Identification and characterization of an Epstein-Barr virus-specific T-cell response in the pathologic tissue of a patient with Hodgkin's disease.
abstract: Several lines of evidence indicate that an impairment of EBV-specific immune responses may contribute to the pathogenesis of Hodgkin's disease (HD). At present, however, it is not clear whether a defective immunity to EBV is a characteristic restricted to EBV-associated HD cases or a more generalized phenomenon, part of the inherent immune deficiency of HD patients. In this study, we have addressed this issue by analyzing EBV-specific responses in infiltrating T lymphocytes (TILs) from one HD biopsy, where the virus was confined to a small proportion of apparently normal lymphocytes. TIL cultures were established using low amounts of recombinant interleukin 2 and in the absence of specific stimulation, conditions that preferentially induce the proliferation of in vivo activated T cells. An EBV-specific cytotoxic component was revealed by the capacity of these TILs to lyse autologous EBV-positive lymphoblastoid cell lines (LCLs) obtained by spontaneous transformation from the lesion but not HLA-mismatched LCLs and autologous phytohemagglutinin blasts. This cytotoxic activity closely resembled that of EBV-specific memory T cells, which may be reactivated from the blood lymphocytes of healthy donors by in vitro stimulation with autologous LCLs. The use of a panel of appropriately HLA-matched B95.8-transformed LCLs as targets in standard 51Cr release assays revealed EBV-specific cytotoxic responses to be restricted mainly through the A11 and B44 HLA alleles with a minor HLA-A26-restricted component. Using autologous fibroblasts infected with recombinant vaccinia viruses expressing the EBV latent antigens, the TIL culture was shown to recognize latent membrane protein 2 and, to a lesser extent, EBV-encoded nuclear antigen 6. In addition, a strong proliferative response was induced by coculture of TILs with autologous but not with allogeneic LCLs or autologous phytohemagglutinin blasts. Six CD4-positive, EBV-specific T-cell clones were isolated by limiting dilution. The study of cytokine mRNA expression, carried out by reverse transcriptase-assisted PCR, revealed that three of these T-cell clones expressed a Th0 phenotype, whereas 1 had a Th2 phenotype. These findings are consistent with the presence in this HD lesion of an ongoing immune response against EBV-carrying cells and suggest that the complex immune deficiency that characterizes HD patients probably does not include a generalized, constitutional defect of EBV-specific T-cell responses.
authors.affiliation: Division of Experimental Oncology, Centro di Riferimento Oncologico, Aviano PN, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Dolcetti
authors.forename: T
authors.initials: T
authors.lastname: Frisan
authors.forename: J
authors.initials: J
authors.lastname: Sjöberg
authors.forename: P O
authors.initials: PO
authors.lastname: De Campos-Lima
authors.forename: P
authors.initials: P
authors.lastname: Pisa
authors.forename: V
authors.initials: V
authors.lastname: De Re
authors.forename: A
authors.initials: A
authors.lastname: Gloghini
authors.forename: S
authors.initials: S
authors.lastname: Rizzo
authors.forename: M G
authors.initials: MG
authors.lastname: Masucci
authors.forename: M
authors.initials: M
authors.lastname: Boiocchi
mesh_terms: D000328:Adult; D000956:Antigens, Viral / Q000276:immunology*; D001483:Base Sequence; D016207:Cytokines / Q000235:genetics; D003602:Cytotoxicity, Immunologic; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006689:Hodgkin Disease / Q000276:immunology* / Q000382:microbiology; D006801:Humans; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D000956:Antigens, Viral; D016207:Cytokines; D017931:DNA Primers; D004279:DNA, Viral; D012333:RNA, Messenger