title: Retroviral transduction of protein kinase C-gamma into tumor-specific T cells allows antigen-independent long-term growth in IL-2 with retention of functional specificity in vitro and ability to mediate tumor therapy in vivo.
abstract: The aim of our study was to examine the potential usefulness of transducing the protein kinase C-gamma (PKC-gamma) cDNA gene into tumor-specific T cells as a technique for facilitating the generation of large numbers of functional Ag-specific T for tumor therapy. Murine CD8+, F-MuLV gag-specific CTL clones, and CD4+, F-MuLV env-specific Th clones, as well as bulk-cultured T cell lines with defined Ag specificity to FBL-3, a Friend murine leukemia virus (F-MuLV)-induced tumor, were transduced with a retroviral vector pZipNeoPKC-gamma and selected in G418. The results demonstrated that PKC-gamma-transduced clones remained activated in culture, as evidenced by continued expression of up-regulated levels of IL-2R, which were as high after 6 mo in culture without Ag restimulation as 24 h after Ag stimulation. In vitro functional studies demonstrated that PKC-gamma-transduced CD8+ T cell clones maintained specific cytolytic activity to FBL-3, and PKC-gamma-transduced CD4+ T cell clones maintained specific proliferative activity to FBL-3 or F-MuLV Ag presented by irradiated syngeneic APC. Short-term bulk-cultured T cells specific to FBL-3 were also transduced and could be grown long term in vitro with maintenance of functional specificity. In vivo study showed that PKC-gamma-transduced CD4+ T cells were able to proliferate in response to Ag plus IL-2 stimulation in vivo in a similar pattern as the parental T cells. Therapy with adoptively transferred PKC-gamma-transduced T cell clones and lines into syngeneic mice, with or without FBL-3 tumor, showed that the PKC-gamma-transduced T cells were not tumorigenic and were effective in curing mice with disseminated FBL-3.
authors.affiliation: Department of Medicine, University of Washington, Seattle 98195.
authors.forename: W
authors.initials: W
authors.lastname: Chen
authors.forename: E
authors.initials: E
authors.lastname: Schweins
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: O J
authors.initials: OJ
authors.lastname: Finn
authors.forename: M A
authors.initials: MA
authors.lastname: Cheever
mesh_terms: D000818:Animals; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002455:Cell Division; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007111:Immunity, Cellular; D007167:Immunotherapy; D007376:Interleukin-2 / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics*; D004915:Leukemia, Erythroblastic, Acute / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000235:genetics*; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D015854:Up-Regulation
chemical_list: D017931:DNA Primers; D007376:Interleukin-2; D007527:Isoenzymes; D015375:Receptors, Interleukin-2; C087857:protein kinase C gamma; D011493:Protein Kinase C