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目的观察旋毛虫70 ku重组热休克蛋白(rTs-Hsp70)对体外培养的小鼠骨髓源树突状细胞(BMDC)的活化作用。方法分离BMDC,体外培养至第6 d,培养基中加入rTs-Hsp70刺激培养24 h,ELISA检测培养上清中细胞因子IL-12P70和TNF-α的含量,流式细胞仪检测BMDC表面标志分子CD11c和CD86的表达,同时观察细胞的形态。实验平行设立不加刺激物的阴性对照组,细菌脂蛋白LPS阳性对照组和煮沸变性的rTs-Hsp70组。结果 rTs-Hsp70刺激组DC培养上清中的IL-12P70和TNF-α水平与不加刺激物对照比较差异有统计学意义(P<0.05);CD86+双阳性细胞的百分比为27.0%;电镜观察rTs-Hsp70刺激后的DC呈成熟细胞形态,相邻细胞间的突起形成连接。结论 rTs-Hsp70可能通过诱导DC的成熟而使其活化,从而激活小鼠产生抗旋毛虫感染的免疫保护作用。
Objective To investigate the activation of mouse bone marrow-derived dendritic cells (BMDC) cultured in vitro by 70 ku recombinant heat shock protein (rTs-Hsp70). Methods BMDC was isolated and cultured on day 6 in vitro. The cells were stimulated with rTs-Hsp70 for 24 h. The contents of cytokines IL-12P70 and TNF-α in culture supernatants were detected by ELISA. The expression of BMDC surface marker CD11c and CD86 expression, while observing the cell morphology. In the experiment, a negative control group without stimulus, a bacterial lipoprotein LPS positive control group and a boiled denatured rTs-Hsp70 group were set up in parallel. Results The levels of IL-12P70 and TNF-α in the supernatant of DCs stimulated with rTs-Hsp70 were significantly different from those of the control without stimulation (P <0.05). The percentage of CD86 + double positive cells was 27.0% DCs stimulated by rTs-Hsp70 showed mature cell morphology, and the protrusions of adjacent cells formed connections. Conclusion rTs-Hsp70 may be activated by inducing the maturation of DC, thereby activating mice to produce the anti-Trichinella infection immune protection.