【摘要】 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。更多还原

【Abstract】 Objective To observe if 70 ku recombinant heat shock protein from Trichinella spiralis(rTs-Hsp70) activates murine bone marrow-derived dendritic cells(BMDCs) in vitro.Methods Murine BMDCs were isolated and cultured in vitro.After five days of culturing,BMDCs were cultured together with rTs-Hsp70 for 24 hours.The cytokines IL-12P70 and TNF-α,which were produced in BMDC culture supernatants stimulated with rTs-Hsp70 in vitro,were detected by ELISA.The levels of CD11c and CD86 were examined with flow cytometry.At the same time,the morphologies of BMDCs were observed.BMDCs cultured with LPS,with denatured rTs-Hsp70,and without stimulation served as controls.Results The level of IL-12P70 or TNF-α in BMDCs stimulated with rTs-Hsp70 differed statistically from BMDCs without stimulation(P<0.05).The percentage content of CD86+ double-positive cells in BMDCs stimulated with rTs-Hsp70 was 27.0%.Electron microscopy revealed that BMDCs stimulated with rTs-Hsp70 had a mature morphology and that projections from adjacent cells formed a link between cells.Conclusion rTs-Hsp70 activates DCs by stimulating their maturation,and this finding provides clues about the mechanism of action of rTs-Hsp70 in the immune response to T.spiralis.更多还原