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目的探讨B7-H1阻断对未成熟树突状细胞(DCs)的免疫刺激活性的影响。方法以细胞因子GM-CSF和IL-4体外诱导人单核细胞来源的树突状细胞,流式细胞仪检测在诱导过程中B7-H1的表达,并以B7-H1单抗阻断处理,分别以流式细胞仪、MTT法、ELISA、ELISPOT法检测B7-H1阻断对DCs的成熟和内吞能力、刺激淋巴细胞增殖、IL-12的分泌、诱导淋巴细胞分化的影响。结果DCs在诱导过程中B7-H1表达逐渐增强,第7d时的阳性表达率为54.12%,以TNF-α诱导成熟后阳性表达率为83.64%;B7-H1阻断对DCs成熟和内吞能力无影响,但可使未成熟DCs刺激淋巴细胞增殖的能力和IL-12的分泌明显增强,并诱导淋巴细胞向分泌IFN-γ的Th1/Tc1的分化。结论B7-H1阻断可增强未成熟DCs的免疫刺激活性,利用B7-H1阻断的DCs瘤苗激发抗肿瘤免疫的方案值得进一步探讨。
Objective To investigate the effect of B7-H1 block on immunostimulatory activity of immature dendritic cells (DCs). Methods Human monocyte-derived dendritic cells were induced with cytokines GM-CSF and IL-4 in vitro. The expression of B7-H1 was detected by flow cytometry and blocked by B7-H1 monoclonal antibody. The effects of B7-H1 block on the maturation and endocytosis of DCs, the proliferation of lymphocytes, the secretion of IL-12 and the induction of lymphocyte differentiation were detected by flow cytometry, MTT assay, ELISA and ELISPOT respectively. Results The expression of B7-H1 in DCs gradually increased during the induction of DCs. The positive expression rate of DCs was 54.12% on the 7th day, and the positive expression rate was 83.64% after induced by TNF-α. The B7-H1 blocked the maturation and endocytosis of DCs But did not affect the ability of immature DCs to stimulate lymphocyte proliferation and the secretion of IL-12, and induced the differentiation of lymphocytes to Th1 / Tc1 secreting IFN-γ. Conclusions B7-H1 blockade can enhance the immunostimulatory activity of immature DCs. The anti-tumor immune response induced by B7-H1 DCs vaccine should be further explored.