Santin A D, Ioli G R, Hiserodt J C, Rose G S, Graf M R, Tocco L M, Lander J K, Eck L M, Burger R A, DiSaia P J
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717, USA.
Gynecol Oncol. 1995 Aug;58(2):230-9. doi: 10.1006/gyno.1995.1216.
Human ovarian carcinoma cell lines were genetically engineered to secrete the cytokine interleukin-4 (IL-4) by retroviral-mediated gene transduction. These cells were transduced with the LXSN retroviral vector containing the human IL-4 gene and the neomycin resistance selection marker. Numerous IL-4-secreting clones were isolated from different papillary serous carcinoma cell lines, including SKOV-3, UCI-101, and UCI-107, and one clone derived from UCI-107 extensively characterized. This clone, termed UCI 107E IL-4 GS, was shown to constitutively express high levels of IL-4 (i.e., 900 to 1300 pg/ml/10(5) cells/48 hr) for over 35 passages and 6 months of study. Like the parental cell line (UCI-107), UCI 107E IL-4 GS cells expressed MHC class I and Her-2/neu surface antigens but did not express detectable MHC class II, ICAM 1, CA 125, or IL-4 receptors. No increase in expression of surface proteins was noted between parental and UCI 107E IL-4 GS. The morphology of this clone did not differ from that of the parental or LXSN vector control cells; however, parental cells had a faster growth rates than transductants. UCI 107E IL-4 GS was sensitive to gamma irradiation since as little as 2500 rad killed most of the cells within 10 days of irradiation. However, after irradiation, IL-4 secretion continued until about Day 8. The potential use of these IL-4-secreting ovarian carcinoma cells as vaccines for woman with advanced ovarian cancer will be discussed.
通过逆转录病毒介导的基因转导，对人卵巢癌细胞系进行基因工程改造，使其分泌细胞因子白细胞介素-4（IL-4）。这些细胞用含有人类IL-4基因和新霉素抗性选择标记的LXSN逆转录病毒载体进行转导。从不同的乳头状浆液性癌细胞系中分离出许多分泌IL-4的克隆，包括SKOV-3、UCI-101和UCI-107，并对一个源自UCI-107的克隆进行了广泛表征。这个克隆被称为UCI 107E IL-4 GS，在超过35代和6个月的研究中，显示出持续高水平表达IL-4（即900至1300 pg/ml/10(5)个细胞/48小时）。与亲本细胞系（UCI-107）一样，UCI 107E IL-4 GS细胞表达MHC I类和Her-2/neu表面抗原，但未检测到表达MHC II类、ICAM 1、CA 125或IL-4受体。亲本细胞与UCI 107E IL-4 GS之间未观察到表面蛋白表达增加。该克隆的形态与亲本或LXSN载体对照细胞的形态没有差异；然而，亲本细胞的生长速度比转导细胞快。UCI 107E IL-4 GS对γ射线照射敏感，因为低至2500拉德的照射在照射后10天内杀死了大多数细胞。然而，照射后，IL-4分泌一直持续到大约第8天。将讨论这些分泌IL-4的卵巢癌细胞作为晚期卵巢癌女性疫苗的潜在用途。
