Nayak S K, McCallister T, Han L J, Gangavalli R, Barber J, Dillman R O
Cell Biology Laboratory, Hoag Cancer Center, Newport Beach, CA 92663, USA.
Cancer Gene Ther. 1996 May-Jun;3(3):143-50.
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.
我们使用了一种含有人类γ-干扰素（γ-IFN）基因的逆转录病毒载体来转导13种肾癌细胞系。通过使用含有LacZ标记基因的类似载体测定，转导效率范围为0%至60%。此外，利用对抗生素新霉素的基因转移抗性来筛选转导细胞。13个细胞系中有9个成功转导。转导与细胞伸长的形态学变化相关，并且细胞生长速率显著降低。通过酶联免疫吸附测定法测量，转导细胞在转导后至少2至3周（包括转导的第1天、筛选的6 - 7天以及转导细胞首次传代前的另外8 - 12天）分泌不同量（20 - 1076 pg/10(6)细胞/天）的γ-IFN。7个细胞系中的6个细胞系中人类白细胞抗原（HLA）II类表达显著增加；8个细胞系中的2个细胞系中HLA I类表达显著增加。照射后进行冷冻保存的转导细胞仍产生γ-IFN并表达HLA I类和II类抗原，尽管通常水平低于这些操作之前。本研究证实，将人类γ-IFN基因通过逆转录病毒载体转导入肾癌细胞是可行的，并且与γ-IFN的持续产生以及HLA I类和II类分子表达增加相关，并且这些效应在照射和冷冻保存后仍然保留。这表明用照射后的γ-IFN基因转导的肾癌细胞进行自体肿瘤细胞疫苗试验是合理且可行的。
