汉滩病毒核蛋白与热休克蛋白GRP94、HSP27的相互作用

为研究汉滩病毒(Hantaan virus，HTNV)感染诱导乳鼠脑组织热休克蛋白GRP94、HSP27与病毒蛋白的相互关系，选出生2—3d的昆明乳鼠实验性感染汉滩病毒，取8d后发病乳鼠脑组织部分制石蜡切片，用免疫组化结合共聚焦显微镜检测组织中病毒抗原及GRP94、HSP27的表达，部分制匀浆液，用ELISA、免疫共沉淀方法分析病毒抗原和GRP94、HSP27的关系。结果示汉滩病毒感染诱导乳鼠脑组织神经细胞高表达GRP94且与细胞内病毒抗原有共定位关系，但未见HSP27诱导高表达；免疫共沉淀显示汉滩病毒核心抗原(HINV—NP)与GRP94、HSP27呈非共价复合物形式存在。该结果为进一步探讨HSPs在病毒感染复制中的作用以及抗病毒感染方面提供了有意义的实验资料。

The Interaction of Viral Nucleocapsid Protein with GRP94 and HSP27 in Brains of Suckling Mice Experimentally Infected with Hantaan virus

To study the association of heat-shock protein GRP94 and HSP27 with Hantaan virus nucleocapsid protein (HTNV-NP) during viral replication, new-born mice less than 72h of age were experimentally infected with Hantaan virus intracerebrally. Brains of mice at the 8th day post-infection were prepared for tissue sections and extracts. Hantaan virus infection, GRP94, HSP27 expression and their localization were demonstrated by immunohistochemical staining. The possible interaction between HTNV-NP and GRP94, HSP27 was analyzed by ELISA, immunoprecipitation. HTNV infection led to increased expression of GRP94 but not HSP27 at cytoplasma of neurons in brains of infected suckling mice, colocalizationof GRP94 with HTNV-NP, and association of both GRP94 and HSP27 with HTNV-NP. The results provide the basis for further research of the roles of HSPs in the viral infection and replication.