Ohira T, Ohe Y, Heike Y, Podack E R, Olsen K J, Nishio K, Nishio M, Miyahara Y, Funayama Y, Ogasawara H
Pharmacology Division, National Cancer Center Research Institute, Tokyo, Japan.
Gene Ther. 1994 Jul;1(4):269-75.
Gene therapy with cytokine cDNA will provide a new tool for cancer treatment. We have already reported that immunization with interleukin-2 (IL2) cDNA transfected Lewis lung carcinoma (LLC) cells induced anti-tumor immunity, which, however, was not strong enough to eradicate an established tumor. In an attempt to develop more effective gene therapy methods, we have used tumor cells co-transfected with IL-2 and tumor necrosis factor (TNF) cDNAs. These cDNAs were introduced into pBMG-Neo and pcDV-X819 vectors, respectively, and then co-transfected into LLC cells. The co-transfectants were selected by incubating them in a medium containing G418 followed by the limiting dilution method twice to obtain IL2 and TNF cDNA co-transfected LLC (LLC-TNF-IL2) cells. When 5 x 10(5)/ml LLC-TNF-IL2 cells were incubated for 48 h, they secreted 7.56 U/ml TNF and 527.0 U/ml IL2 into the culture supernatant. When C57BL/6 mice were transplanted with 1 x 10(6) LLC-TNF-IL2 cells, all the tumors were rejected. The growth of transplanted LLC, but not B16F10 melanoma cells, was retarded in mice inoculated with LLC-TNF-IL2 on their contralateral sides, which suggests specific immunity was induced. The immunization effect by the co-transfectant was superior to that of the IL2- and TNF-transfectants alone.
用细胞因子cDNA进行基因治疗将为癌症治疗提供一种新工具。我们已经报道，用白细胞介素-2（IL2）cDNA转染的Lewis肺癌（LLC）细胞进行免疫可诱导抗肿瘤免疫，然而，这种免疫强度不足以根除已形成的肿瘤。为了开发更有效的基因治疗方法，我们使用了共转染了IL-2和肿瘤坏死因子（TNF）cDNA的肿瘤细胞。这些cDNA分别被导入pBMG-Neo和pcDV-X819载体，然后共转染到LLC细胞中。通过在含有G418的培养基中培养共转染细胞，然后采用有限稀释法进行两次筛选，以获得IL2和TNF cDNA共转染的LLC（LLC-TNF-IL2）细胞。当5×10⁵/ml的LLC-TNF-IL2细胞培养48小时时，它们向培养上清液中分泌7.56 U/ml的TNF和527.0 U/ml的IL2。当将1×10⁶个LLC-TNF-IL2细胞移植到C57BL/6小鼠体内时，所有肿瘤均被排斥。在对侧接种LLC-TNF-IL2的小鼠中，移植的LLC细胞的生长受到抑制，但B16F10黑色素瘤细胞不受影响，这表明诱导了特异性免疫。共转染细胞的免疫效果优于单独的IL2和TNF转染细胞。
