日本血吸虫16kDa蛋白调节DC高表达IL-10的信号途径及其分子机制研究

"Schistosome immune evasion" mechanism is an important theoretical problem of prevention and control of Schistosomiasis. Among these, initiative regulation of host immune response by releasing some schistosome-derived molecules is an important mechanisms of immune evasion of Schistsoma japonicum. Sj16 is a new gene that our group identified in Schistosoma adult cDNA library at 1999, and then we have performed a series of work for this gene. Previous studies have shown that rSj16 has a clearly immune modulation effect for the host. The common characteristic of rSj16 on immune cells was that rSj16 can production a large number of IL-10, but the mechanism was not clear yet. This project intends to use immunohistochemistry, real-time quantitative PCR and Western-blot analysis methods and PRRs knockout mice model to screen and verified the key PRRs related to IL-10 signal pathways in dendrite cells stimulated by rSj16 and recognize the mechanisms of this PRRs promotes Interleukin 10 production in dendritic cells stimulated by rSj16. The objective of this study was to reveal the immunity mechanism of regulation of host immune responses by rSj16 and illuminate the molecule mechanism of immune evasion of Schistosoma japonicum, which can rich the understanding of mechanisms of relationship of Schistosome and host and lay a basic experiment foundation for the guidance of schistosomiasis vaccine research and anti-schistosomiasis drug development.

"血吸虫免疫逃避"机制是血吸虫病防治研究的重要理论问题。通过释放虫源性物质主动调节宿主的免疫应答，是血吸虫免疫逃避的重要方式。Sj16是申请者所在课题组克隆到的日本血吸虫新基因，具有确切的免疫调节作用，对多种免疫细胞作用的共同的特征是诱导大量的IL-10的产生，但其作用机制尚不明确。本项目拟利用免疫组化、实时定量PCR、Western blot等方法，对rSj16诱导小鼠BMDC高表达IL-10产生的相关的PRRs进行筛选和验证；并利用PRRs缺陷小鼠模型对筛选到的PRRs与rSj16调节BMDC表达IL-10的相关性进行验证；同时利用过表达和RNAi等技术初步探讨PRRs介导的rSj16调节DC高表达IL-10的机制。本研究旨在深入揭示日本血吸虫感染免疫的特征和免疫逃避的分子机制，以期为揭示宿主与寄生虫关系，为指导血吸虫病疫苗研究及抗血吸虫病药物开发奠定新的实验基础。

“血吸虫免疫逃避”机制是血吸虫病防治研究的重要理论问题。通过释放虫源性物质主动调节宿主的免疫应答，是血吸虫免疫逃避的重要方式。Sj16是申请者所在课题组克隆到的日本血吸虫新基因，具有确切的免疫调节作用，对多种免疫细胞作用的共同的特征是诱导大量的IL-10的产生，但其作用机制尚不明确。本项目利用IL-10抗体及IL-10R抗体和IL-10(-/-)小鼠探讨rSj16负调DC功能与IL-10之间的关系，证实rSj16负调LPS的BMDC功能是IL-10直接依赖的并通过IL-10R通路来发挥作用；利用TLR4 KO小鼠和TLR2 KO小鼠评价了PRRs在rSj16负调DC诱导IL-10升高方面的关系；同时发现rSj16具有两段核定位序列，rSj16利用NLS59-79引导rSj16靶向进入BMDC细胞核内，是rSj16发挥负调LPS诱导的BMDC上调IL-10表达功能的关键序列。本项目的成功开展，对于深入揭示日本血吸虫感染免疫的特征和免疫逃避的分子机制，以期为揭示宿主与寄生虫关系，为指导血吸虫病疫苗研究及抗血吸虫病药物开发奠定新的实验基础。目前已经发表SCI论文2篇，投稿1篇，并申请1项发明专利。