摘要
目的研究热休克蛋白70(HSP70)-肝癌抗原肽(SMMC7721人肝癌细胞株)复合物修饰的树突状细胞(DC)对细胞因子诱导的杀伤(CIK)细胞的增殖活性及表型的影响。方法获取健康供者的外周血单个核细胞(PBMC),培养2 h,贴壁细胞诱导分化DC,非贴壁细胞用于诱导培养CIK细胞,DC生长第7日开始负载HSP70(A组)、肝癌抗原肽(B组)、HSP70-肝癌抗原肽复合物(C组),另设单纯DC组(D组),流式细胞检测CD80、CD86的表达,体外构建HSP70-抗原肽复合物-DC瘤苗,并将其和CIK细胞按1∶10的比例混合培养5 d,四甲基偶氮唑蓝法检测细胞增殖能力,以流式细胞仪检测DC及CIK细胞膜表面分子表达。结果 HSP70-肝癌抗原肽复合物可促进DC成熟,DC表面分子CD86、CD80在C组表达均较D组高(P均<0.01)。HSP70-肝癌抗原肽复合物-DC瘤苗可明显刺激CIK细胞的增殖。HSP70-肝癌抗原肽复合物-DC瘤苗与CIK共培养可使CIK细胞CD3^+CD8^+、CD3^+CD56^+的表达提高,与单纯培养CIK细胞比较差异均有统计学意义(P均<0.05)。结论 HSP70-肝癌抗原肽复合物能提高DC表面分子CD80、CD86等表达,诱导DC的成熟,HSP70-抗原肽复合物-DC瘤苗与CIK细胞共培养可明显增加CIK细胞的增殖活性,提高CIK细胞CD3^+CD8^+、CD3^+CD56^+的表达。
Objective To evaluate the effect of dendritic cells(DC) modified by heat shock protein70(HSP70)-SMMC7721 tumor peptide complex upon the proliferation activities and phenotype changes of cytokine-induced killer(CIK) cells.Methods Peripheral blood mononuclear cells were obtained from healthy donors and cultured for 2 h.The adherent cells were induced to differentiate DC and non-adherent cells were utilized to induce CIK cells.DCs were loaded with HSP70(group A),hepatoma antigen peptide(group B) and HSP70-hepatoma antigen peptide complex(group C) on the seventh day of growth,and DC alone were allocated into group D.The expression of CD80 and CD86 was detected by flow cytometry.HSP70-antigen peptide complex-DC tumor vaccine was constructed in vitro,and co-cultured with CIK cells at a ratio of 1:10.The proliferation of CIK was measured by MTT assay.The expression of molecules on the membrane surface of DCs and CIK was observed by flow cytometry.Results HSP70-SMMC7721 peptide complex could accelerate the maturity of DC.The expression levels of CD80 and CD86 on the membrane surface of DC in group C were significantly higher compared with those in group D(both P<0.01).HSP70-antigen peptide complex-DC tumor vaccine could remarkably stimulate the proliferation of CIK cells.The expression levels of CD3’CD8’,CD3^+CD56^+in CIK cells were significantly up-regulated after the co-culture of HSP70-antigen peptide complex-DC tumor vaccine and CIK compared with those after the culture of CIK cells alone(all P<0.05).Conclusions HSP70-SMMC7721 tumor peptide complex can up-regulate the expression levels of CD80 and CD86 on the membrane surface of DC and induce the maturity of DC.The co-culture of HSP70-antigen peptide complex-DC tumor vaccine and CIK cells can significantly enhance the proliferation activity and up-regulate the expression levels of CD3^+CD8^+,CD3^+CD56^+in CIK cells.
作者
潘洁
宋慧东
王巧瑜
萧间开
Pan Jie;Song Huidong;Wang Qiaoyu;Xiao Jiankai(Department of Gastroenterolgy,Guangzhou No.12 Hospital,Guangzhou 510620,China)
出处
《新医学》
2020年第2期117-120,共4页
Journal of New Medicine
基金
广州市医药卫生科技项目基金资助课题（20161A011041）
