Rab32通过调节自噬参与树突状细胞抗胞内细菌免疫应答的机制研究

Intracellular bacterial infection is currently facing major infectious diseases, serious threating to human health. Innate immunity as well as adaptive immunity is involved in the response of the host towards pathogens. Dendritic cells (DCs) are professional antigen presenting cells playing a central role in immune response by linking the innate and adaptive immunity. Previously studies have shown that in the intereaction of DC and pathogen, changing in the vesicle system may determine the outcome of the anti- bacterial immunity and infection outcome. In our study, we first found that Rab32 is a key molecule in regulating intracellular vesicle transport ; then we found that Rab32 participate in the autophagysome formation, reducing the survival of Listeria monocytogenes. To further explore the mechanisms, first we use ultra-high-resolution microscope to observe the process of Rab32 involved in autophagysome formation; protein network maps are depicted using phagosome proteomics to understand the molecular mechanisms in Rab32 regulating autophagysome; finally, in vivo animal experiments are carried out to observe the effect of anti- bacterial immune response regulated by Rab32.This study helps to understand the role of DC in anti- bacterial immunity, providing a possible target molecules for the development of anti- bacterial drugs and has important theoretical significance and potential applications .

胞内细菌感染是目前面临的重要感染性疾病，严重威胁人类健康。树突状细胞（dendritic cells，DCs）作为天然免疫和获得性免疫的桥梁在宿主抗胞内细菌免疫中地位独特。近年研究显示在DCs与胞内病原菌相互作用过程中，内囊泡系统的改变可能决定了抗细菌免疫的结局和感染转归。在前期发现Rab32是调节胞内囊泡转运关键分子之基础上，我们进一步发现Rab32参与DCs自噬体的形成，降低胞内细菌的存活率。为了探索其机理，本课题拟采用超高分辨率显微镜观察Rab32参与调节DCs自噬的具体环节；通过吞噬体蛋白质组分析Rab32调节自噬的蛋白网络图，了解Rab32调节自噬的分子机制；在体动物实验观察Rab32抗细菌免疫应答效果；最终阐明Rab32通过自噬调节DCs抗胞内细菌感染的作用机制。本研究有助于深入了解DCs在抗细菌免疫中的作用，为发展抗细菌药物提供可能的靶分子，具有重要的理论意义和应用价值。

胞内细菌感染是目前面临的重要感染性疾病，严重威胁人类健康。树突状细胞（dendritic cells，DCs）作为天然免疫和获得性免疫的桥梁在宿主抗胞内细菌免疫中地位独特。近年研究显示在DCs与胞内病原菌相互作用过程中，内囊泡系统的改变可能决定了抗细菌免疫的结局和感染转归。在前期发现Rab32是调节胞内囊泡转运关键分子之基础上，我们进一步发现Rab32参与DCs自噬体的形成，降低胞内细菌的存活率。为了探索其机理，我们首先通过串联亲和层析结合LC/MS/MS，鉴定出Rab32吞噬李斯特菌前后的相互作用蛋白网络图；其次发现以Rab32/PHB1/PHB2蛋白复合物为标志的囊泡，参与限制李斯特菌在DC细胞的胞内复制；再次通过超高分辨率显微镜和电镜发现限制李斯特菌胞内复制囊泡性质部分为自噬囊泡；最后通过在体动物实验观察，敲除Rab32后李斯特菌的存活情况，通过体内和体外实验最终阐明Rab32通过Rab32/PHB1/PHB2调节DCs抗胞内细菌感染的作用机制。本研究有助于深入了解DCs在抗细菌免疫中的作用，为发展抗细菌药物提供可能的靶分子，具有重要的理论意义和应用价值。