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从ｐｃＤ－５．１质粒和ｐＳＰＧＮＨＩＦＮＢ１０质粒分别切下人白细胞介素２（ＨｕＩＬ－２）ｃＤＮＡ和人β干扰素（ＨｕＩＦＮ－β）ｃＤＮＡ，分别克隆到ｐＮＭＳＭ逆转录病毒载体多克隆位点，经酶切鉴定确定正确插入方向。经ＰＡ３１７包装细胞包装成重组病毒，测转染效率。用ＮＩＨ／３Ｔ３细胞测重组病毒感染效率。在８μｇ／ｍｌ鱼精蛋白存在的条件下分别感染人新鲜肾透明细胞癌体外建株细胞以及人肾细胞癌肿瘤浸润性淋巴细胞（ＴＩＬ）。经Ｇ４１８选择阳性克隆后测上清表达ＨｕＩＬ－２及ＨｕＩＦＮ－β活性。结果ＨｕＩＬ－２和ＨｕＩＦＮ分别在转ＨｕＩＬ－２基因和ＨｕＩＦＮ－β基因肾细胞癌细胞的表达量达２５０Ｕ／１０６细胞·ｄ－１和６４００Ｕ／１０６细胞·ｄ－１，在经目的基因转染ＴＩＬ细胞中的表达量分别比未经目的基因转染的ＴＩＬ高２～３倍和４～８倍。ＴＩＬ生长状况未有明显改变，抗肿瘤活性未见明显增强。
The human interleukin 2 (HuIL-2) cDNA and human interferon beta (HuIFN-β) cDNA were cut from pcD-5.1 plasmid and pSPGNHIFNB10 plasmid and cloned into pNMSM retroviral vector polyclonal site respectively. Digestion identification to determine the correct insertion direction. The PA317 packaging cells packaged into recombinant virus, transfection efficiency. Recombinant virus infection efficiency was measured using NIH / 3T3 cells. In vitro culture of human renal clear cell carcinoma cells and human renal cell carcinoma tumor infiltrating lymphocytes (TIL) were respectively infected in the presence of 8μg / ml protamine. After the positive clones were selected by G418, the expression of HuIL-2 and HuIFN-β was detected. Results The expression levels of HuIL-2 and HuIFN were up to 250 U / 106 cells · d-1 and 6400 U / 106 cells · d-1 respectively in HuIL-2 and HuIFN-β cells. TIL cells were expressed 2 to 3 times and 4 to 8 times higher than the TILs transfected with the gene of interest respectively. TIL growth status did not change significantly, no significant increase in anti-tumor activity.