摘要
目的探讨人HER2阳性乳腺癌细胞对树突状细胞表型及功能的影响。方法分离培养外周血单个核细胞来源的树突状细胞。培养的第5天,实验组HER2+MCF-7细胞的培养上清液孵育24小时,以不加上清液的树突状细胞作为对照组,流式细胞术检测树突状细胞表面标志CD80、CD86、CD40、CD83和HLA-DR的表达水平,ELISA法检测树突状细胞细胞因子IL-12、TNF-α、IL-6和IL-10的分泌水平。用改良的MTT比色法检测上清液处理的树突状细胞刺激同种异体T细胞增殖的能力和ELISA法检测树突状细胞刺激T细胞分泌细胞因子的能力。结果树突状细胞与HER2+MCF-7细胞的上清液共培养后,树突状细胞表面分子CD80、CD83、CD86及CD40均表达升高,同时伴随着IL-12、TNF-α、IL-6的分泌增加。与对照组相比较,上清液处理的树突状细胞对激活同种异体T细胞增殖和分泌细胞因子的能力明显增强(P<0.01)。结论 HER2+MCF-7细胞能改善树突状细胞的抗原递呈功能和刺激T细胞活化增殖的能力,提示HER2+MCF-7细胞对树突状细胞是免疫刺激作用,对于HER2+的乳腺癌可采用基于树突状细胞疫苗的免疫治疗。
Objective To explore the effects of HER2+MCF-7 cell supernatant on phenotype and function of dendrit- ic cells. Methods The peripheral blood mononuclear cells (PBMCs) were isolated and then cultured in RPMI-1640 with recombinant GM-CSF (10 ng/mL), IL-4 (50 ng/mL) to obtain dendritic cells (DCs). On day 5 of culture, DCs were treat- ed with HERUMCF-7 for 24 h. Supernatant- untreated DCs were used as control group. The expression of co-stimulato- ry molecules CDS0, CD86, CD40, CD83 and HLA-DR in DCs was evaluated by FACS, IL-12, TNF- α and IL-6 produc- tion by DCs was examined using ELISA. The proliferation and secretion of allogenetic T cells stimunlated by Supernatant- treated DCs were evaluated by MTT and ELISA. Results The levels of co-stimulatory molecules CDS0,CD86, CD40, CD83 and HLA-DR were elevated in DCs, accompanied by increased production of IL-12, TNF- c~, IL-6. Moreover, compared with the control group,HER2 + MCF- 7- exposed DCs stimulated allogenetic T cell proliferation and T cell- derived cytokine production(P〈0.01). Conclusions These results indicate that HERU MCF-7 can improve DCs' anti- gen presenting function and capability to activate T cells, suggesting that HER2+ MCF-7 cells may have an immunostimu- latory effect on DCs, and DC based immunotherapy may be adopted in the treatment of breast cancer.
出处
《菏泽医学专科学校学报》
2014年第4期8-11,共4页
Journal of Heze Medical College
作者简介
孙伟红,女,(1974-),博士,主管技师。研究方向:肿瘤免疫、感染免疫和免疫调节的研究。
通讯作者:高岱清,E—maihgao—daiqing@163.com。