人树突状细胞与CD137单抗联合体外抗宫颈癌的实验研究

Study of anti-cervical carcinoma effect in vitro mediated by human dendritic cells combined with CD137 McAb

目的 :研究树突状细胞 (DCs)联合CD137单抗对HeLa宫颈癌细胞系体外生长的抑制作用。方法 :外周血单核细胞体外经细胞因子GM CSF、IL 4的诱导获得DCs ,流式细胞术检测DCs相关表型及协同刺激分子CD137的表达。MTT法分别检测DCs以及DCs联合CD137单抗对HeLa细胞的生长抑制作用。结果 :诱导 5～ 7d后的悬浮细胞具有典型的树突状细胞形态。流式细胞术检测结果显示 ,MHCⅡ类分子HLA DR的表达率为中等水平 ,单核 巨噬细胞系表面标志CD14表达率小于 5 % ;CD137的表达率为 9.77% ,LPS诱导成熟后CD137的表达率增加为 36 .0 6 %。当DCs与HeLa细胞的效靶比为 1.2 5 :1、2 .5 :1、5 :1、10 :1时 ,DCs对HeLa细胞的抑瘤率分别为 6 .17%、5 .75 %、2 3.4 6 %、2 1.4 8% ;当效靶比为5 :1时 ,DCs联合CD137单抗抑制HeLa细胞生长的作用 ,比DCs单独抑制HeLa细胞生长的作用明显增强 ,其抑瘤率由 2 5 .4 4%提高到 5 7.0 7% (P <0 .0 5 )。结论 :DCs联合CD137单抗 ,体外具有显著的抗宫颈癌作用 ,为宫颈癌的防治提供了新思路。

Objective:To study the growth-inhibition of HeLa cervix carcinoma cell line mediated by human dendritic cells(DCs) combined with CD137 monoclonal antibody(McAb) in vitro.Methods:DCs were induced from the human peripheral blood by human recombinant granulocyte-macrophage clony stimulating factor (GM-CSF) and interlukin-4(IL-4) in vitro.The related phenotypes of DCs and the expression of CD137 on DCs were detected by flow cytometry.The growth-inhibition of HeLa cell mediated by human DCs as well as by DCs combined with CD137 McAb were detected by MTT respectively.Results:After 5～7 days' culture, the non-adherent cells had the typical morphology of DCs.Flow cytometry showed these cells expressed 64.02% of HLA-DR and 2.34% of the monocyte-macrophage lines special marker CD14.Human immature DCs expressed 9.77% of CD137,otherwise on human mature DCs stimulated by lipopolysaccharide(LPS) the expression of CD137 increased to 36.06%.When effector to target ratios of DCs to HeLa cells were 1.25:1,2.5:1,5:1,10:1,the growth-inhibition rates of HeLa cells mediated by human DCs were 6.17%,5.75%,23.46%,21.48% respectively.When effector to target ratio was 5:1, the growth-inhibition of HeLa cells mediated by human dendritic cells combined with CD137 McAb enhanced(from 25.44% to 57.07%).Conclusion:Human DCs combined with CD137 McAb have significant anti-cervical carcinoma effect, which may provide a new idea for prevention and therapy of cervix carcinoma.