论文部分内容阅读
目的:探讨人外周血γδT细胞体外诱导过程中CD107a的表达变化及其与γδT细胞细胞毒活性的关系。方法:分离健康人外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),加入含有IL-2和异戊烯焦磷酸(isopentenyl pyro-phosphate,IPP)的培养基,体外诱导γδT细胞。分别在第7、10、14天以流式细胞仪对培养的γδT细胞进行鉴定,并同时检测其CD107a、穿孔素和颗粒酶B的表达。CCK-8试剂盒检测γδT细胞对胰腺癌细胞SW-1990的杀伤效应。采用SPSS13.0软件进行spearman相关分析。结果:在培养的第7、10、14天,γδTCR阳性细胞分别为(60.31±3.84)%、(66.45±4.25)%、(70.99±4.66)%。CD107a、穿孔素和颗粒酶B的表达在γδT细胞培养第7~10天达到峰值后呈下降趋势,第7天与第0天相比差异有统计学意义[(80.66±4.42)%、(70.11±3.34)%、(94.26±4.25)%vs(69.02±5.04)%、(62.31±4.66)%、(53.62±3.69)%,P<0.05]。培养第7天、第10天的γδT细胞对SW-1990细胞杀伤率显著高于第14天的γδT细胞[(58.86±5.12)%、(61.53±4.69)%vs(40.31±4.83)%,P<0.05]。γδT细胞CD107a表达与穿孔素、颗粒酶B、其对SW-1990细胞杀伤效应显著相关(r=0.853,r=0.785,r=0.839,均P<0.01)。结论:人外周血γδT细胞CD107a的表达与其抗肿瘤能力正相关,可能是γδT细胞细胞毒活性的一个标志。
OBJECTIVE: To investigate the changes of CD107a expression in human peripheral blood γδT cells in vitro and their relationship with the cytotoxic activity of γδT cells. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers. The medium containing IL-2 and isopentenyl pyro-phosphate (IPP) was added to induce γδT cells in vitro. The cultured γδT cells were identified by flow cytometry on the 7th, 10th, and the 14th day respectively, and the expressions of CD107a, perforin and granzyme B were detected simultaneously. CCK-8 kit was used to detect the killing effect of γδT cells on pancreatic cancer cells SW-1990. SPSS13.0 software was used for spearman correlation analysis. Results: At day 7, day 10 and day 14, the positive rate of γδTCR positive cells were (60.31 ± 3.84)%, (66.45 ± 4.25)% and (70.99 ± 4.66)%, respectively. The expression of CD107a, perforin and granzyme B decreased after the 7th to 10th day of γδT cell culture, and the difference was statistically significant on the 7th day and the 0th day [(80.66 ± 4.42)%, (70.11 ± 3.34%, (94.26 ± 4.25)% vs (69.02 ± 5.04)%, (62.31 ± 4.66)%, (53.62 ± 3.69)%, P <0.05. The killing rate of γδT cells on day 7 was significantly higher than that on day 14 [(58.86 ± 5.12)%, (61.53 ± 4.69)% vs (40.31 ± 4.83)%, P <0.05]. The expression of CD107a in γδT cells was significantly correlated with the killing effect of perforin and granzyme B on SW-1990 cells (r = 0.853, r = 0.785, r = 0.839, all P <0.01). Conclusion: The expression of CD107a in human peripheral blood γδT cells is positively correlated with its anti-tumor ability, which may be a sign of the cytotoxic activity of γδT cells.