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自从Rosenberg报道以来,对各种癌应用LAK过继性免疫疗法虽引人注目,但尚未用自体肝癌细胞系研究过肝癌细胞对LAK的敏感性。作者用本研究室建立的分泌白蛋白的肝癌培养细胞株Hep-Tabata和该建株的肝癌病人的外周血单核细胞(PBMC)研究了LAK细胞活性。方法:①NK活性的测定:将Hep-Tabata作为自体系靶细胞,将SK-Hep-1、Hep-3B作为同种异体性靶细胞,将K_(562)、Raji作为对照靶细胞。按规则用~(51)Cr标记上述靶细胞,每孔滴布5×10~3个靶细胞。将该肝癌病人和正常人的全血中分离出的PBMC(用Ficoll-Paque法分离),按80:1、40:1、20:1的E/T之比分别加入各孔,37℃、5%CO_2中培养12小时。培养后沉淀,将所得的培养上清液用穴式闪烁计算器测定,算出细胞毒性值。②LAK细胞活性的测定:在LAK细胞浓度调节到2×10~8/ml的PBMC中加入IL-2(IU/ml),37℃、5%CO_2中培养4天。结果:①该肝癌病人PBMC和正常人PBMC均能
Since Rosenberg’s report, the application of LAK adoptive immunotherapy to various cancers has attracted attention. However, the sensitivity of hepatoma cells to LAK has not been studied in autologous hepatoma cell lines. The authors studied the activity of LAK cells using peripheral blood mononuclear cells (PBMCs) of hepatocellular carcinoma cultured cell line Hep-Tabata and human hepatocellular carcinoma cells established in this laboratory. Methods: Determination of 1NK activity: Hep-Tabata was used as a target system cell, SK-Hep-1 and Hep-3B were used as allogeneic target cells, and K 562 and Raji were used as control target cells. The above target cells were labeled with ~(51)Cr according to the rule, and 5×10~3 target cells were dripped per well. The PBMCs isolated from the whole blood of the liver cancer patients and normal persons (separated by the Ficoll-Paque method) were added to each well according to the E/T ratio of 80:1, 40:1, and 20:1, respectively, at 37C. Incubate for 12 hours in 5% CO 2 . After the culture, the precipitate was precipitated, and the obtained culture supernatant was measured using a hole scintillation counter to calculate the cytotoxicity value. 2 Measurement of LAK cell activity: IL-2 (IU/ml) was added to PBMCs with LAK cell concentration adjusted to 2×10~8/ml, cultured at 37° C., 5% CO 2 for 4 days. Results: 1 Both PBMC and normal human PBMC of this liver cancer patient can