模拟失重对IL-2诱导的人NK细胞生物活性的影响  

Effects of simulated weightlessness on biological activity of human NK cells induced by IL-2

作　　者：刘文利[1,2] 朱霞[2] 赵莉[1] 杨喜玲[1] 曹菲[2] 黄勇[3] 穆沛红[1] 

机构地区：[1]西北农林科技大学医院,陕西杨凌712100 [2]第四军医大学医学心理系,陕西西安710032 [3]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《细胞与分子免疫学杂志》2015年第10期1297-1300,1305,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：西北农林科技大学国际科技合作基金(A213021310)

摘　　要：目的明确模拟失重对白细胞介素2(IL-2)诱导的人自然杀伤(NK)细胞活性的影响。方法回转细胞模拟失重条件培养人NK细胞,CCK-8法检测NK细胞增殖活性、异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞凋亡率、ELISA检测γ干扰素(IFN-γ)产生水平,反转录PCR检测NK细胞IL-12β1和IL-12β2受体mRNA水平。结果与正常重力组相比,模拟失重培养24 h和48 h后,IL-2诱导的NK细胞增殖水平分别下降13.6%和31%,凋亡率分别增加8%和19%,IFN-γ分泌水平分别降低25.2%和47.8%,杀伤活性分别下降7%和18%,IL-12刺激IL-2预处理NK细胞产生IFN-γ的水平分别降低21.8%和58.8%,同时,NK细胞IL-12β1和IL-12β2受体mRNA水平显著降低。结论模拟失重抑制IL-2诱导的NK细胞增殖、IFN-γ产生和杀伤活性,并进一步通过下调IL-12受体mRNA表达抑制IL-12诱导的IFN-γ产生。Objective To investigate the effects of simulated weightlessness on the activity of human natural killer （NK） cells induced by interleukin 2 （IL-2）. Methods Primary human NK cells were cultured under simulated weightlessness condition. The viability of NK cells was determined by CCK-8 assay; cell apoptosis was analyzed by flow cytometry combined with annexin V-FITC/PI staining; the level of interferon y （IFN-γ） was examined by ELISA; the mRNA levels of IL-12 receptor genes were detected by reverse transcription PCR. Results Compared with control cells cultured in normal gravity, IL-2-induced cell proliferation rate of NK cells cultured in simulated weightlessness decreased by 13.6% and 31% at 24 and 48 hours, respectively; the cell apoptotic rate increased by 8% and 19% ; IL-2-induced IFN-y production was inhibited by 25.2% and 47.8%; the cytotoxicity of NK cells induced by IL-2 was reduced by ？% and 18%; IL-12-induced IFN-y production was suppressed by 21.8% and 58.8% in IL-2 pretreated cells at 24 and 48 hours, respectively. In addition, the mRNA levels of IL-12 β1 and β2 receptor genes were significantly down-regulated in the cells cultured in simulated weightlessness. Conclusion Simulated weightlessness can inhibit the proliferation of NK cells induced by IL-2, promote NK cell apoptosis, impair IL-2-induced IFN-y production and cytotoxicity in NK cells, and inhibit IL-12-induced IFN-γ production through down-regulating IL-12 receptor gene expression in NK cells.

关 键 词：模拟失重 自然杀伤细胞 细胞杀伤活性 Γ干扰素 

分 类 号：R852.22[医药卫生—航空、航天与航海医学] Q254[医药卫生—临床医学]