IL-18诱导外周血单个核细胞对Raji细胞的杀伤效应及其机制的实验研究

目的 探讨IL-18诱导PBMC对Raji细胞的杀伤效应及其作用机制。方法 IL-18在体外诱导PBMC后,以MTT法测定其对Raji细胞的杀伤效应。在杀伤活性最大时,以BAS-ELISA法检测培养上清液sFas-L的含量,以流式细胞仪检测PBMC的CD_(16)/CD_(56)、Fas-L、穿孔素、颗粒酶B表达。结果 PBMC在体外经IL-18诱导后,能明显增强其对Raji细胞的杀伤活性,在PBMC经IL-18诱导后的d 4,这种杀伤活性最高。在杀伤活性最高时,PBMC表达CD16/CD_(56)、Fas-L、穿孔素、颗粒酶B的能力明显增加,但培养上清液中sFas-L的含量无明显增加。结论 本实验的结果显示,IL-18能诱导PBMC杀伤细胞的增殖和活化,并使其对Raji细胞杀伤活性增强,这种杀伤机制可能与穿孔素、颗粒酶B和Fas-L的高表达有关。

The experimental study of human peripheral blood mononuclear cells killer activity and its mechanisms induced by interleukin-18 to raji cell line

AIM To investigate the PBMC cy-totoxicity induced by IL-18 to Raji cell and the cy-totoxicity mechanisms. METHODS In vitro PBMC was induced by IL-18, then the cytotoxicity of the induced PBMC to Raji cell line was detected by MTT assay. When the cytotoxicity was the highest, the content of sFas-L from culture medium was measured by BAS-ELISA method, and the expressions of CD16/CD56, perforin, granzyme B, Fas-L deriving from induced PBMC were detected by flow cytometer. RESULTS The cytotoxicity of PBMC induced by IL-18 to Raji cells enhanced with days going. At the fourth day, the cytotoxicity was the highest. And at that time, the expressions of CDis/CDse, perforin, granzyme B, Fas-L deriving from PBMC increased, but the sFas-L content from culture medium did not increase. CONCLUSIONS IL-18 could promote the proliferation of cytotoxic cells deriving from PBMC and augment the cytotoxicity of the PBMC to Raji cells. The cytotoxicity mechanisms might relate to high expressions of perforin, granzyme B and Fas-L.