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目的建立并优化阿萨希毛孢子菌临床株(Trichosporon asahii,T.asahii)与野生型小鼠骨髓来源树突状细胞(Bone marrow-derived dendritic cell,BMDC)共培养体系。方法体外培养C57BL/6小鼠BMDC。用不同浓度比例的热灭活T.asahii与BMDC共培养,筛选最优浓度比例,用脂多糖(LPS)和β-葡聚糖(β-glucan)分别刺激BMDC,选择适宜阳性对照;利用优化的体系进一步检测BMDC表面共刺激分子的激活和相关细胞因子的产生情况。结果 BMDC与T.asahii比例为1∶5时最适宜作为共培养的比例,而β-glucan更适于作为阳性对照;小鼠BMDC能够对T.asahii进行抗原提呈,其通过增加IL-6和TNF-α的产生发挥抗真菌作用。结论该共培养体系能够有效检测小鼠BMDC对T.asahii的免疫效应。
Objective To establish and optimize the co-culture system of Trichosporon asahii (T.asahii) and wild-type mouse bone marrow-derived dendritic cells (BMDC). Methods C57BL / 6 mice BMDC were cultured in vitro. The co-culture of T.asahii and BMDC was induced by heat inactivation at different concentrations, and the optimum concentration was screened. BMDC was stimulated with lipopolysaccharide (LPS) and β-glucan respectively to select suitable positive control. System to further detect the activation of costimulatory molecules on BMDC surface and the related cytokine production. Results The ratio of BMDC to T.asahii was best at 1: 5, and β-glucan was more suitable as a positive control. Mouse BMDC was capable of antigen presenting T.asahii by increasing IL-6 And TNF-α production play an anti-fungal effect. Conclusion The co-culture system can effectively detect the immune effects of mouse BMDC on T.asahii.