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目的探讨Bryostatin-1对树突状细胞(DC)免疫功能的调节作用。方法联合应用GM-CSF和IL-4自人外周血单核细胞定向分化DC;以Bryostatin-1刺激TNF-α诱导成熟的DC,收集DC及其上清夜,以FACS和ELISA方法分析DC表面免疫分子(CD80、CD83、CD86、HLA-DR和B7-H1)和细胞因子(IFN-γ、IL-1β、IL-10和IL-12)表达水平;自DC提取RNA,以Northern blot分析DC的B7-h1 mRNA表达水平;以DC为诱导细胞,进一步检测DC的免疫诱导功能。结果Bryostatin-1通过降低DC表面B7-H1表达和增强B7.2表达,以及调节DC细胞因子(IL-1b、IL-12和IFN-γ)分泌,增强DC的免疫诱导功能,PKC通道特异性的抑制剂BI明显逆转Bryostatin-1的上述作用。结论Bryostatin-1增强DC免疫功能,其机制可能是激活PKC通道。
Objective To investigate the regulatory effect of Bryostatin-1 on immune function of dendritic cells (DCs). Methods DCs were differentiated from human peripheral blood mononuclear cells by GM-CSF and IL-4. DCs induced by TNF-α were stimulated by Bryostatin-1, DCs and their supernatants were collected and analyzed by FACS and ELISA (CD80, CD83, CD86, HLA-DR and B7-H1) and cytokines (IFN-γ, IL-1β, IL-10 and IL-12); RNA was extracted from DCs and analyzed by Northern blot B7-h1 mRNA expression levels; DC induced cells, further detection of DC immune induction function. Results Bryostatin-1 enhanced the immune induction function of DC by down-regulating the expression of B7-H1 and enhancing the expression of B7.2 on the DC surface as well as the secretion of DC cytokines (IL-1b, IL-12 and IFN- Inhibitor BI significantly reversed the above effects of Bryostatin-1. Conclusion Bryostatin-1 enhances the immune function of DC, and its mechanism may be to activate PKC channel.