知识与财富的链接
目的探讨rhIL-21对NK-92MI细胞免疫功能的影响。方法以不同浓度(25～100ng/ml)的rhIL-21培养NK-92MI细胞24～72h,观察NK-92MI细胞的增殖。以50ng/ml的rhIL-21处理NK-92MI不同时间后,FACS检测NK-92MI细胞的凋亡,NK-92MI细胞表面NKG2D受体及胞内颗粒酶-B蛋白的表达;细胞杀伤试剂盒检测NK-92MI细胞对靶细胞K-562的细胞毒效应;RT-PCR检测NK-92MI受体NKG2D,效应分子颗粒酶-B、穿孔素及细胞因子IFN-γ基因mRNA的表达;ELISA检测其分泌IFN-γ的水平。结果 RhIL-21虽能抑制NK-92 MI细胞的增殖(P<0.05),但不会促进细胞的凋亡(P<0.05)。RhIL-21能促进NK-92MI细胞穿孔素、IFN-γ、颗粒酶-B、NKG2D受体基因及相应蛋白质的表达上调(P<0.05)。rhIL-21预处理组NK-92MI对K-562细胞的细胞毒效应也明显增强(P<0.05)。结论 RhIL-21能增强NK-92 MI细胞的细胞毒效应,IL-21在肿瘤的免疫治疗中有潜在临床应用价值。
Objective To investigate the effect of rhIL-21 on NK-92MI cells.Methods Using in vitro cultural experiments,NK-92MI cells were treated with different concentration of rhIL-21 and then the proliferation were tested at different time.In addition,after NK-92MI cells treated with 50ng/ml rhIL-21 for 48h or 72h,the NK-92MI cell activity,apoptotic cells,the expressions of NKG2D and granzyme-B was measured using FACS assay.The levels of IFN-γfrom the supernatants were detected by ELISA.The expressions of NKG2D,perforin,granzyme-B,and IFN-γ mRNA were determined by RT-PCR.Results RhIL-21 inhibited NK-92MI cell proliferation.However,rhIL-21 did not affect cell apoptosis.RhIL-21 increased the expressions of the effector molecules granzyme-B and IFN-γ as well as activation receptor NKG2D at the mRNA and protein levels.Furthermore,compared with control,rhIL-21 enhanced the cytotoxicity of NK92-MI cells against K-562 target.Conclusion RhIL-21 could enhance the cytotoxicity of NK92-MI cells possibly by up-regulating the expression of activation and effector molecular,suggesting that IL-21 might have therapeutically effect to enhance NK cell-mediated antitumour responses.
