【摘要】 目的:研究透明质酸对小鼠骨髓来源树突状细胞功能的影响以及回输后荷黑色素瘤小鼠脾淋巴细胞增殖、活化和细胞因子的变化,进而探讨透明质酸诱导的树突状细胞增强荷瘤小鼠免疫功能的机制。方法:体外细胞因子联合诱导培养小鼠骨髓细胞获得树突状细胞(DCs),免疫磁珠分选纯化获得CD11c+树突状细胞,经不同浓度透明质酸(HA)刺激后,采用酶联免疫吸附法(ELISA)检测培养上清液中细胞因子IL-12p70含量。建立小鼠皮下B16黑色素瘤模型,肿瘤局部皮下回输HA孵育DC后检测肿瘤大小,应用Con A检测脾淋巴细胞增殖情况,应用MTT法检测脾淋巴细胞杀伤活性,ELISA法检测脾淋巴细胞分泌的TNF-α和IFN-γ的表达,以单纯DC回输、生理盐水注射以及正常小鼠(无瘤)组作为对照。结果:在10~100μg/m L范围内,HA以剂量依赖的方式上调DCs分泌IL-12p70。HA孵育DC处理组肿瘤生长明显受到抑制;淋巴细胞增殖反应、杀伤活性和细胞因子TNF-α和IFN-γ的表达明显高于单纯DC组和生理盐水组(P<0.05)。结论:透明质酸可促进小鼠骨髓DC的成熟;透明质酸孵育的DC通过增强荷瘤小鼠的抗肿瘤免疫功能而抑制肿瘤的生长。更多还原

【Abstract】 Objective: To investigate the effects of immune function of dendritic cells(DCs) mediated by Hyaluronic Acid(HA)on the transplanted B16 melanoma in mice. Methods: Bone marrow-derived dendritic cells were induced by culture supplemented with recombinant mouse GMSF and IL-4, and then CD11c+cells isolated by MACS were cultured for 24 h in the presence of HA at varying concentrations.The levels of IL-12p70 in culture supernatant of DCs were detected by ELISA.Tumor size was measured after treatment to evaluate the reaction of the treatment. Con A methods were used to observe splenic lymphocytes proliferation. Splenic lymphocytes were also extracted to assess the cytotoxic activity against B16 melanoma cells by MTT assay. Furthermore, both TNF-αand IFN- γ levels in the supernatant of culture were detected by ELISA after a 24- hour culture of spleen lymphocytes with B16 melanoma cells. Results: HA up-regulated the secretion of IL-12p70 in a dose-dependent manner at 10-100 μg /m L. Growth of the tumor was significantly inhibited in group treated with dendritic cells mediated by Hyaluronic Acid as compared with the controls. The lymphocytes proliferation, lymphocytes cytotoxic activity and TNF-α and IFN-γ levels in group treated with dendritic cells mediated by Hyaluronic Acid were significantly higher than that of the controls with statistical significance(P< 0.05). Conclusions: HA may play a role in inducing the maturation of DCs. The treatment with dendritic cells mediated by Hyaluronic Acid could inhibit the growth of the tumor by improve the host anti-tumor immune reaction.更多还原