B7-1分子增强NK细胞对肝癌细胞毒活性的体外研究

In vitro study on the role of B7-1 molecule in cytotoxicity of natural killer cells against hepatocellular carcinoma

目的：了解Ｂ７１ 分子在体外抗肝癌免疫反应中对自然杀伤细胞（ＮＫ 细胞） 杀伤活性的影响。方法：经脂质体ＤＯＳＰＥＲ 介导将ｈＢ７１ 基因直接导入ＨｅｐＧ２ 细胞，建立ＨｅｐＧ２／ｈＢ７１ 细胞克隆。分离健康人和肝癌患者外周血淋巴细胞，以ＭＴＴ 法测定ＮＫ 细胞对ＨｅｐＧ２／ｈＢ７１ 细胞的杀伤活性，以杀伤ＨｅｐＧ２／ｎｅｏ 和ＨｅｐＧ２ 细胞为对照。结果： ＨｅｐＧ２ 细胞转染ｈＢ７１ 基因后表达Ｂ７１ 分子。无论在健康人组还是肝癌患者组， 健康人外周血ＮＫ 细胞杀伤ＨｅｐＧ２／ ｎｅｏ ， ＨｅｐＧ２ 细胞的活性约为肝癌患者ＮＫ 活性水平的２ 倍。健康人和肝癌患者外周血ＮＫ 细胞对ＨｅｐＧ２／ｈＢ７１ 细胞的杀伤活性均较对照组明显增高（ 增强１－４９～３－４８ 倍） ， 对ＨｅｐＧ２／ ｈＢ７１ 细胞的杀伤活性在Ｅ／ Ｔ＝ ５ ∶１时分别为５７－２ 及４３－８ ，有显著性差异（ Ｐ＜ ０－０５） ，在Ｅ／Ｔ＝ １０ ∶１ 及２０ ∶１ 时分别可达８２－８ ～８４－４ 和７８－６ ～８６－４ ，差异不显著（ Ｐ ＞ ０－０５） 。结论：Ｂ７１ 分子显著增强ＮＫ 细胞对肝癌细胞的体外杀伤活性，可能通过激活非特异性细胞免疫在抗肝癌复发中发挥作用。

Objective:To investigate the effect of B7 1 expression on cytotoxicity of human natural killer cells (NK cells) against hepatocellular carcinoma (HCC) cells in vitro. Methods:Human B7 1 gene was directly transduced into HepG2 cells in presence of DOSPER to establish HepG2/hB7 1 cell clone. Peripheral blood lymphocytes (PBL) were seperated from healthy donors and HCC patients blood and subjected to test NK cytotoxicity by detecting their ability to lyse HepG2,HepG2/neo and HepG2/hB7 1 cells by MTT dye method. Results:HepG2/hB7 1 cell clone was established through direct B7 1 gene transfer,which strongly expressing B7 1 molecules. NK cytotoxicity against HepG2,HepG2/neo in healthy donorsPBL was about 2 times much as that of in HCC patientsPBL. NK cytotoxicity in healthy donors and HCC patients PBL against HepG2/hB7 1 cells were 2 23～3 48 and 1 49～2 14 times higher than that of lysing HepG2/neo cells,respectively (P< 0 001). NK cytotoxicity ability in HCC patients PBL against HepG2/hB7 1 cell was 43 8,lower than that of healthy donors(57 2) at E/T=5∶1,while E/T=10∶1,20∶1,they reached 78 6～86 4,as much as the same of healthy donors 82 8～84 4, no statistical significance was found (P >0 05). Conclusions:B7 1 molecules strongly enhance NK cytotoxicity against hepatocellular carcinoma cells in vitro.