摘要
目的探讨CD28单克隆抗体对人外周血γδT细胞体外增殖及杀伤胃癌BCG823、SGC7901细胞活性的影响。方法分离健康人外周血单个核细胞(PBMC),分别在有或无CD28单抗(20μg/ml)参与的条件下,加入到含帕米膦酸、IL-2的RPMI1640完全培养基中诱导培养γδT细胞。在培养的第10天用台盼蓝染色计数细胞;采用流式细胞术检测各组γδT细胞的纯度及其穿孔素、颗粒酶B和CD107a的表达;用CCK-8试剂盒检测γδT细胞对胃癌细胞BCG823的体外杀伤效应。结果在诱导培养10d后,两组γδT细胞纯度均达到70%以上,与无CD28单抗对照组相比,CD28单抗组γδT细胞纯度显著高于对照组;CD28单抗组γδT细胞扩增倍数、穿孔素、颗粒酶B和CD107a的表达及对胃癌细胞BCG823和SGC7901的杀伤活性均显著高于无CD28单抗对照组。结论 CD28共刺激能增强γδT细胞增殖并通过上调穿孔素和颗粒酶B的表达增强其抗肿瘤活性。
Objective To investigate the enhanced effect of anti-CD28 mAb （monoclonal antibody） on proliferation and killing activity of -,/ST cells against gastric carcinoma cells including BCG823 cells and SGC7901 cells in vitro. Methods yQT cells were generated in vitro by stimulating peripheral blood mononu- clear cells of healthy donors with pamidronate and IL-2 at the condition with or without anti-CD28 mAb （20ug/ml） respectively. yQT cell number in each group were countered by trypan blue staining on the 10th day after culture. The purity of yQT cells and their perforin, granzyme B, IFN-3, and CD107a expression were veri- fied by flow cytometry. The killing activity of yQT cells against BCG823 cells were analyzed by CCK-8 kit. Results The experimental results showed that purity of yQT cells in two groups all exceeded by 70% after 10- day culture. Comparing with anti-CD28 mAb free control group, cell purity and expansion folds of yQT cells in anti-CD28 mAb group were significantly higher than those in control group. Moreover, perforin, granzyme B, CD107a expression and cytotoxicity against BCG823 cells and SGC7901 cells of yQT cells with anti-CD28 mAb group were significantly higher than those of the control group. Conclusion These results indicate that CD28 costimulation could promote yQT cells proliferation and enhance yQT cells mediated antitumor activity by up-regulation of pefforin, granzyme B, and IFN-y expression.
出处
《现代消化及介入诊疗》
2013年第1期1-4,共4页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
南京军区医学科技创新(08MA039)
