SELDI-TOF-MS检测胎盘免疫调节因子对CIK细胞蛋白谱的影响

Effect of placental immunoregulatory factor detected by SELDI-TOF-MS on protein profiling of CIK-cell

目的用表面加强激光解析电离飞行时间质谱技术(SELDI-TOF-MS)检测胎盘免疫调节因子(placental factor,PF)作用后的人细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK细胞)的蛋白组变化。方法将健康人新鲜血液诱导成为CIK细胞后,设PF-CIK组和不加PF的空白对照CIK组,继续培养72h,观察CIK细胞的增殖率和细胞毒活性的变化,并用表面加强激光解析电离飞行时间质谱技术(SELD-ITOF-MS)及WCX2蛋白芯片获得CIK细胞蛋白质指纹图谱,用计算机软件进行比较分析,寻找差异蛋白。结果与空白对照CIK细胞组相比较,PF-CIK组细胞的增殖率和杀伤率均有统计学意义(P<0.01)。空白对照CIK组和PF-CIK组有6个峰差异有统计学意义(P<0.05),其中以4个蛋白质生物标志物(M/Z分别为5636.077、6860.359、6072.814和5984.191)差异最大。结论PF可上调CIK细胞的生长及杀伤活性,并从蛋白组学上分析可发现两组之间存在显著差异的蛋白峰。同时,SELDI-TOF-MS在CIK细胞特异性蛋白质生物标志分子的筛选等方面具有一定价值。

Objective SELDI technology was used to detect the changes of protein expression in cytokine-induced killer cells （CIK） which were culture in the medium contains placental factor （PF）. Methods Peripheral blood mononuclear cells （PBMNCs） were collected from healthy human donors. After being induced as eytokine induced-killer （CIK） ,the cells were divided into two groups. The CIK group was cultured with 10% FBS culture media only, and the PF-CIK group was cultured with 10% FBS culture media that was added placental factor （PF）. The proteomic spectra of CIK were obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS） on WCX-2 chips. The differences of proteins between CIK group and PF-CIK group were compared. Results The results of SELDI showed that a total of 6 protein was significandy different （P〈0.05）, especially the four with molecular weight as 5636.077, 6860.359, 6072.814 and 5984. 191. Condusions PF might have the function that can change the expression level of CIK. SELDI technology can find out the differentially expressed protein in CIK effectively.