Notch信号通路在DC介导的细粒棘球蚴重组抗原Eg.ferritin免疫过程中的作用研究  

The role of the Notch signaling pathway in DC-dependent immunity of Echinococcus granulosus ferritin

作　　者：吕士玉 高富[1,2] 李子华[2,3] 王强[2,3] 赵巍[2,4] 王娅娜[2,3,4] 

机构地区：[1]宁夏医科大学检验学院,宁夏银川750004 [2]宁夏医科大学医学科学研究所 [3]宁夏医科大学基础学院 [4]宁夏医科大学细胞生物学与遗传学教研室

出　　处：《中国病原生物学杂志》2014年第12期1098-1103,共6页Journal of Pathogen Biology

基　　金：宁夏回族自治区自然科学基金项目(No.NZ13224)

摘　　要：目的初步探索Notch信号对树突状细胞(dendritic cell,DC)介导的抗寄生虫感染免疫反应的影响。方法体外培养骨髓来源的小鼠DC细胞,利用γ-分泌酶抑制剂N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine tbutyl ester,DAPT于不同时间段处理细胞,阻断DC的Notch信号通路,培养第7d时收集DC,用细粒棘球蚴重组抗原铁蛋白(ferritin protein of Echinococcus granulosus,Eg.ferritin)体外刺激DC,利用扫描电镜观察DC形态的改变,采用流式细胞术检测DC表面分子MHCⅡ、CD40及CD80/CD86的表达情况。结果 DC细胞数量随DAPT浓度的增高而降低;经不同浓度DAPT于不同时间段处理后,DC细胞中Notch1mRNA的表达受到抑制,以50μmol/L DAPT在第0d处理DC,对Notch1mRNA的抑制效果更显著(P<0.05);Eg.ferritin和LPS均能刺激DC细胞表面树突的生成及表面分子MHCⅡ、CD40及CD80/CD86的表达,且Eg.ferritin比LPS的刺激作用更显著(P<0.05),而加入DAPT后,DC细胞表面树突的生成减少,各表面分子的表达水平降低(P<0.05),并削弱了Eg.ferritin和LPS的刺激作用。结论Eg.ferritin能促进DC细胞的分化成熟,而阻断Notch信号通路会影响DC细胞的分化成熟,并降低DC细胞对Eg.ferritin的反应能力。其分子机制涉及Notch信号通路。Objective To identify the role of the Notch signaling pathway in the dendritic cell（DC）-mediated immune response to parasites. Methods Theγ-secretase inhibitor DAPT was used to block the Notch signaling of monocyte-derived murine DCs at different times.On day 7,cells were harvested and stimulated with Echinococcus granulosus recombinant ferritin.DC morphology and surface molecule expression were examined using electron microscopy and flow cytometry.DCs were exposed to E.granulosus ferritin,and the immune response of DCs treated with DAPT and not treated with DAPT was compared. Results The number of DCs decreased as the concentration of DAPT increased.On day 0,50μmol/L of DAPT resulted in the greatest inhibition of DCs（P〈0.05）.When DCs were stimulated with E.granulosus ferritin and LPS,they tended to be mature with more dendrites and expressed more surface molecules.Compared to LPS,E.granulosus ferritin was more capable of stimulating dendrite generation and surface molecule expression（P〈0.05）.However,these phenomena were absent in DCs treated with DAPT and stimulated with E.granulosus ferritin and LPS. Conclusion The Notch signaling pathway may affect the differentiation and maturation of DCs,and blocking of the Notch signaling pathway may impair DC-based anti-parasite immunity.E.granulosus ferritin significantly stimulated DC growth.When stimulated with E.granulosus ferritin,Notch signaling-deficient DCs produced a weak immune response.These results suggest that the Notch signaling pathway may be related to DC-based anti-parasite immunity.

关 键 词：NOTCH信号通路 DAPT 细粒棘球蚴重组抗原Eg.ferritin 树突状细胞(DC) 

分 类 号：R383.3[医药卫生—医学寄生虫学]