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目的利用痤疮丙酸杆菌(P.acnes)引起的炎症反应,促进小鼠外周血中树突状细胞(DC)的动员并研究P.acnes动员的DC在体外对胃癌细胞的作用。方法C578BL/6J(B6)小鼠注射P.acnes后外周血分离单个核细胞,用流式细胞仪分选F4/80-B220-CD11c+细胞,在体外加入细胞因子GM-CSF、IL-4和TNFα共培养,通过形态学观察、表型分析和混合淋巴细胞反应鉴定它们是否能分化为成熟DC。将P.acnes动员的DC负载胃癌抗原后致敏T细胞,观察活化的T细胞在体外对胃癌细胞的杀伤效应。结果注射P.acnes1h后外周血F4/80-B220-CD11c+细胞数量即开始升高,24h后逐渐达到高峰。新鲜分离的细胞不具有成熟DC的特征。与细胞因子共培养后即具有典型的DC形态和表型,在混合淋巴细胞反应中具有极强的刺激T细胞增殖的能力。P.acnes动员的DC负载胃癌抗原后致敏T细胞对胃癌细胞的杀伤率明显高于未致敏T细胞。结论P.acnes可迅速动员F4/80-B220-CD11c+细胞进入小鼠外周血,经细胞因子的诱导后可分化为成熟DC,并可在体外诱导出针对胃癌细胞的特异性杀伤T淋巴细胞,对胃癌细胞有明显的杀伤作用,并伴有高水平的INF-γ分泌。
Objective To improve the mobilization of dendritic cells (DCs) in mice peripheral blood by the inflammatory reaction induced by P. acnes and to study the effect of P. acnes-mobilized DC on gastric cancer cells in vitro. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from C578BL / 6J (B6) mice after P. acnes injection. F4 / 80-B220-CD11c + cells were sorted by flow cytometry. Cytokines GM- TNFα were co-cultured to identify whether they could differentiate into mature DC by morphological observation, phenotypic analysis and mixed lymphocyte reaction. After DC mobilized by P. acnes was loaded with gastric cancer antigen, T cells were sensitized and the killing effect of activated T cells on gastric cancer cells in vitro was observed. Results The number of F4 / 80-B220-CD11c + cells in peripheral blood began to increase after 1 h injection of P. acnes, and peaked gradually after 24 h. Freshly isolated cells do not have the characteristics of mature DCs. After co-cultured with cytokines, it has typical DC morphology and phenotype, and has strong ability of stimulating T cell proliferation in mixed lymphocyte reaction. The cytotoxicity of sensitized T cells to gastric cancer cells by P. acnes-mobilized DCs with gastric cancer antigen was significantly higher than that of non-sensitized T cells. Conclusion P. acnes can rapidly mobilize F4 / 80-B220-CD11c + cells into peripheral blood of mice and can differentiate into mature DCs induced by cytokines and induce specific killing T lymphocytes against gastric cancer cells in vitro, On gastric cancer cells have a significant killing effect, accompanied by high levels of INF-γ secretion.