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目的观察肝癌相关抗原kinectin基因重组蛋白(kinectin-MBP)致敏的树突状细胞(DC)对T细胞增殖活化能力的影响。方法从正常人外周血中分离单个核细胞,用rhGM-CSF、rhIL-4联合诱导扩增DC;用kinectin-MBP(kinectin-MBP组)、麦芽糖结合蛋白(MBP组)分别致敏DC,对照组不处理。采用ELISA法检测细胞上清液中的IL-12、IFN-γ,MTT法检测各组DC刺激自体T淋巴细胞增殖的能力。结果kinectin-MBP组上清液中IL-12、IFN-γ含量分别为(615.32±7.93)、(544.28±7.17)pg/ml,MBP组分别为(382.13±5.21)、(308.46±6.44)pg/ml对照组组分别为(299.40±10.22)、(228.03±6.70)pg/ml,三组相比,P均<0.05。kinectin-MBP组T淋巴细胞增殖指数为53.14±0.62,MBP组为22.14±0.31,对照组为10.86±0.65,三组相比,P均<0.001。结论体外诱导扩增的DC经kinectin-MBP致敏后具有很强的刺激自体T细胞增殖的能力。
Objective To observe the effect of dendritic cells (DCs) sensitized by kinectin-MBP on the proliferation and activation of T cells. Methods Mononuclear cells were isolated from normal human peripheral blood and DCs were induced by combination of rhGM-CSF and rhIL-4. DCs were induced by kinectin-MBP and MBP groups, respectively Group does not deal with. IL-12 and IFN-γ in the supernatant of the cells were detected by ELISA. The ability of DCs to stimulate autologous T lymphocyte proliferation was detected by MTT assay. Results The levels of IL-12 and IFN-γ in the kinectin-MBP supernatant were (615.32 ± 7.93) and (544.28 ± 7.17) pg / ml respectively, and those in the MBP group were (382.13 ± 5.21) and (308.46 ± 6.44) pg / ml in the control group were (299.40 ± 10.22) and (228.03 ± 6.70) pg / ml, respectively, P <0.05. Kinectin-MBP group T lymphocyte proliferation index was 53.14 ± 0.62, MBP group was 22.14 ± 0.31, the control group was 10.86 ± 0.65, compared with the three groups, P <0.001. Conclusion DC induced by in vitro stimulation by kinectin-MBP has a strong ability to stimulate autologous T cell proliferation.