title: B lymphoblastoid cell lines as efficient APC to elicit CD8+ T cell responses against a cytomegalovirus antigen.
abstract: Potent and readily accessible APC are critical for development of immunotherapy protocols to treat viral disease and cancer. We have shown that B lymphoblastoid cell lines (BLCL) that stably express CMV phosphoprotein 65 (BLCLpp65), as a result of retroviral transduction, can be used to generate ex vivo CTL cultures that possess cytotoxicity against CMV and EBV. In this report, we demonstrate that the EBV-specific cytotoxicity in the BLCLpp65-primed culture had a spectrum of EBV-Ag recognition similar to that of the BLCL-primed counterpart, suggesting that retroviral transduction and expression of the CMV Ag would not compromise the Ag-presenting capacity of BLCL. In addition, BLCLpp65 appeared to present multiple natural pp65 epitopes, because pp65-specific CTL, which recognized different CMV clinical isolates, were generated in BLCLpp65-primed cultures from individuals with various HLA backgrounds. Consistent with a polyclonal expansion of virus-specific CTL, T cell lines established from the BLCLpp65-primed CTL cultures expressed different TCR-Vbeta Although most of the virus-specific T cell isolates were CD8+, EBV-specific CD4+ lines were also established from BLCLpp65-primed cultures. Western blot analysis revealed that the CD8+ lines, but not the CD4+ line, expressed granzyme B, consistent with features of classic CTL. Thus, our results suggested that BLCL stably expressing a foreign Ag might be used as a practical APC to elicit CD8+ T cell responses.
authors.affiliation: Hematology and Oncology, School of Medicine, University of Alabama, Birmingham 35294, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Sun
authors.forename: R L
authors.initials: RL
authors.lastname: Burton
authors.forename: L J
authors.initials: LJ
authors.lastname: Dai
authors.forename: W J
authors.initials: WJ
authors.lastname: Britt
authors.forename: K G
authors.initials: KG
authors.lastname: Lucas
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D000956:Antigens, Viral / Q000276:immunology* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D016827:CD8 Antigens / Q000096:biosynthesis; D002461:Cell Line, Transformed; D002999:Clone Cells; D003587:Cytomegalovirus / Q000276:immunology* / Q000302:isolation & purification; D003602:Cytotoxicity, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D053804:Granzymes; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation / Q000235:genetics; D018655:Lymphocyte Count; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D054353:Perforin; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D052899:Pore Forming Cytotoxic Proteins; D012697:Serine Endopeptidases / Q000096:biosynthesis; D013602:T-Lymphocytes, Cytotoxic / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D014161:Transduction, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000956:Antigens, Viral; D016827:CD8 Antigens; D018984:Epitopes, T-Lymphocyte; D008562:Membrane Glycoproteins; D010750:Phosphoproteins; D052899:Pore Forming Cytotoxic Proteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D054353:Perforin; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases