CD300LG增强CIK的细胞毒性活性。
CD300LG improves the cytotoxic activity of CIK.
作者信息
Wang Qiuyue, Liu Yuqing, Chen Ying, Wang Ke, Xie Wensheng, Wei Dapeng, Hu Lijuan
机构信息
College of Preclinical and Forensic Medicine, Sichuan University, China.
Department of Immunology, College of Preclinical and Forensic Medicine, Sichuan University, Chengdu, China.
出版信息
Cent Eur J Immunol. 2017;42(2):117-122. doi: 10.5114/ceji.2017.69352. Epub 2017 Aug 8.
To investigate the effect of CD300LG- induction on the cytotoxic activity of CIK. Eukaryotic expression plasmid hCD300LG-/pEGFP-C3, which can express human CD300LG-, was constructed and transfected into CHO cells by lipofectamine. The expression of CD300LG- was confirmed by immunofluorescence, RT-PCR, and Western Blot. To produce CIK cells, human peripheral blood mononuclear cells (PBMC) were isolated and induced, respectively, by cell lysates extracted from hCD300LG-/CHO cells, pEGFP-C3/CHO cells, and CHO cells, concurrently with the standard CIK inductive agent. The cytotoxic activity of these CIK cells against hCD300LG-/CHO cells, pEGFP-C3/CHO cells, CHO cells, and K562 cells was tested. The results showed that eukaryotic expression of plasmid hCD300LG-/pEGFP-C3 was constructed and transfected into CHO cells successfully. After being induced by cell lysates, the cytotoxicity of hCD300LG-/CHO-CIK was improved compared with the other CIK cells. In particular, the activity of killing pEGFP-C3/CHO and CHO cells was improved significantly. Meanwhile, the activity of hCD300LG-/CHO-CIK killing K562 was improved significantly compared with the other CIK cells. The results indicated that hCD300LG- induction can significantly improve the killing activity of CIK cells.
为研究CD300LG诱导对细胞因子诱导的杀伤细胞（CIK）细胞毒性活性的影响。构建了可表达人CD300LG的真核表达质粒hCD300LG-/pEGFP-C3，并通过脂质体转染法将其转染至CHO细胞。通过免疫荧光、逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（Western Blot）证实CD300LG的表达。为制备CIK细胞，分离人外周血单个核细胞（PBMC），分别用从hCD300LG-/CHO细胞、pEGFP-C3/CHO细胞和CHO细胞中提取的细胞裂解物，同时联合标准CIK诱导剂进行诱导。检测这些CIK细胞对hCD300LG-/CHO细胞、pEGFP-C3/CHO细胞、CHO细胞和K562细胞的细胞毒性活性。结果显示，成功构建真核表达质粒hCD300LG-/pEGFP-C3并将其转染至CHO细胞。经细胞裂解物诱导后，hCD300LG-/CHO-CIK的细胞毒性与其他CIK细胞相比有所提高。特别是，其杀伤pEGFP-C3/CHO和CHO细胞的活性显著提高。同时，hCD300LG-/CHO-CIK杀伤K562细胞的活性与其他CIK细胞相比也显著提高。结果表明，CD300LG诱导可显著提高CIK细胞的杀伤活性。
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