原头蚴外泌体对树突状细胞活化和细胞因子分泌的影响

为分析囊型包虫原头蚴(PSC)分泌的外泌体(Exosome)对树突状细胞(DCs)活化和分泌细胞因子的影响,本研究采用高速离心法结合外泌体提取试剂盒分离原头蚴分泌的外泌体(PE),收集的PE沉淀中加入去离子水低渗裂解离心,提取超滤后的蛋白浓缩液即为原头蚴外泌体超滤裂解物(PEL).利用免疫磁珠从健康ICR小鼠脾脏中分选...

Study on the influence of Protoscoleces exosome on the activation and cytokine secretion of dendritic cells

The aim of this study is to analyze the effects of exosome secreted by Protoscoleces(PSC)on the activation and secretion of inflammatory factors in dendritic cells(DCs),and to explore the role of exosome secreted by Protoscoleces in interaction between host and parasite and immune escape Protoscoleces exosome(PE)was separated by high-speed centrifugation.The precipitation of PE was lysed by deionized water and centrifuged,the concentrated liquid extract is PE lysates(PEL).Dendritic cells(DC)were isolated from the spleens of healthy ICR mice by immunomagnetic beads.After 24 hours of stimulation and culture with PE,LPS,PEL and PBS,the cells and culture supernatants were collected.Flow cytometry was used to detect the expression of activated molecules on DC surface and the secretion of cytokines.The result showed that the expression of CD40,CD80,CD86 and MHCⅡon the surface of DCs from PEL group were significantly higher than those of PE group,moreover both of the two groups were significantly higher than those of PBS group(p<0.05),however all PE,PEL and PBS groups s were lower than those of the LPS group.The levels of IL-6,IL-10,IL-12 and TNF-αin DCs culture supernatants of PE and PEL groups were significantly higher than those of PBS group,and the levels of IL-12 and TNF-a in PEL group were higher than those in PE group,but the levels of IL-6 and IL-10 in PEL group were lower than those in PE group.These data showed that PE can stimulate the activation of DCs and release of cytokines.PEL with purified exosome protein and removal of the components of microRNA can induce more effective activation of DCs and promote the release of cytokines,suggesting that exosome protein can induce anti-infective immune response,and exosome microRNA may participate in the immune escape of worms.This study may provide theoretical and experimental data for further investigation on the immunity response of infection and anti-infection of parasites.