Transduction of human renal carcinoma cells with human gamma-interferon gene via retroviral vector.
We used a retroviral vector containing a human gamma-interferon (gamma-IFN) gene to transduce 13 renal carcinoma cell lines. The transduction efficiencies ranged from 0% to 60%, as determined by using an analogous vector containing the LacZ marker gene. In addition, gene-transferred resistance to the antibiotic neomycin was used to select for transduced cells. Nine of 13 lines were successfully transduced. Transduction was associated with the morphologic change of elongation, and there was a marked decrease in cell growth rate. Transduced cells secreted varying amounts (20-1076 pg/10(6) cells/d) of gamma-IFN as measured by enzyme-linked immunosorbent assay for at least 2 to 3 weeks after transduction (including 1 day of transduction, 6-7 days of selection, and an additional 8-12 days before the first passage of the transduced cells). Human leukocyte antigen (HLA) class II expression was markedly increased in six of seven cell lines; HLA class I expression was significantly increased in two of eight lines. Transduced cells that were subjected to cryopreservation after irradiation still produced gamma-IFN and expressed HLA class I and II antigens, although generally at lower levels than before these manipulations. This study confirms that retroviral vector transduction of the human gamma-IFN gene into renal carcinoma cells is feasible and associated with persistent production of gamma-IFN and increased expression of HLA class I and II molecules, and these effects are retained after irradiation and cryopreservation. This suggests that an autologous tumor cell vaccine trial with irradiated gamma-IFN gene-transduced renal carcinoma cell is rationale and feasible.
Nayak
S K
SK
Cell Biology Laboratory, Hoag Cancer Center, Newport Beach, CA 92663, USA.
McCallister
T
T
Han
L J
LJ
Gangavalli
R
R
Barber
J
J
Dillman
R O
RO
D002292:Carcinoma, Renal Cell; D002455:Cell Division; D002460:Cell Line; D015925:Cryopreservation; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006681:HLA-D Antigens / Q000096:biosynthesis; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis*; D007680:Kidney Neoplasms; D007700:Kinetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins; D012190:Retroviridae; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured
D006681:HLA-D Antigens; D015395:Histocompatibility Antigens Class I; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D007371:Interferon-gamma