【摘要】 目的:探讨负载乳腺癌抗原的人脐带血树突状细胞(DCs)分泌的外泌体的抗肿瘤免疫效应。方法:提取新鲜脐血单个核细胞,体外诱导分化为DCs,提取乳腺癌细胞MCF-7肿瘤抗原,并冲击脐血DCs,加入LPS诱导其成熟,利用流式细胞仪检测表型;超速离心法收获负载抗原的脐血DC外泌体;MTT法检测外泌体诱导的T淋巴细胞增殖反应及其激活的CTL对肿瘤细胞的杀伤效应。结果:脐血DC表面标志CD34、CD80、CD86、CD11c阳性细胞数与新鲜脐血单个核细胞相比明显增加(P<0.05);Western-Blot结果显示,负载乳腺癌的脐血DC外泌体携带有MHC-Ⅱ类、CD40、CD80、CD86分子;成熟脐血DC组及负载乳腺癌抗原脐血DC外泌体对同种异体来源的T细胞均有明显促增殖作用;负载乳腺癌抗原的脐血DC及其外泌体激活的CTL均能对乳腺癌细胞MCF-7产生显著地杀伤作用(P<0.05)。且在效靶比100∶1时,外泌体的杀伤作用高于负载抗原的脐血DC(P<0.05)。结论:负载乳腺癌抗原脐血DC分泌大量外泌体,可促进同种异体T细胞增殖,并能诱导特异性抗乳腺癌的免疫效应。更多还原

【Abstract】 Objective:To explore the antitumor immunologic effect of exosomes secreted by human umbilical cord blood dendritic cells(DCs) loaded with breast cancer antigen.Methods:The mononuclear cells were Abstracted from fresh umbilical cord blood,and then they were induced to differentiate to DCs;tumor antigen of breast cancer MCF-7 cells were Abstracted to shock umbilical cord blood DCs,LPS was added to induce maturity of DCs,flow cytometry was used to detect phenotype;supercentrifugation method was used to collect exosomes of DC in umbilical cord blood loading tumor antigen;MTT method was used to detect proliferative reaction of T lymphocytes induced by exosomes and lethal effect of activated CTL on breast cancer cells.Results:The numbers of CD34,CD80,CD86,and CD11c positive DCs in umbilical cord blood were statistically significantly higher than those of fresh mononuclear cells(P<0.05);the results of Western-Blot showed that exosomes of umbilical cord blood DCs loading breast cancer carried MHC-Ⅱ,CD40,CD80,and CD86 cytokines;the exosomes of mature umbilical cord blood DCs and umbilical cord blood DCs loading breast cancer antigen had obvious promoting effects on allogenic T lymphocytes;umbilical cord DCs loading breast cancer antigen and CTL activated by the exosomes had significantly lethal effects on breast cancer MCF-7 cells(P<0.05);when the ratio of effect cells to target cells was 100∶ 1,the lethal effect of exosomes was statistically significantly higher than that of umbilical cord blood DCs loading breast cancer antigen(P<0.05).Conclusion:Umbilical cord blood DCs loading breast cancer antigen secret a great deal of exosomes,which can promote proliferation of allogenic T lymphocytes and induce specific immunological effect of breast cancer.更多还原