摘要目的 证实乳大鼠大脑皮层星形胶质细胞是汉坦病毒属(HV)汉滩病毒(HTNV)和汉城病毒(SEOV)感染的靶细胞,观察病毒感染星形胶质细胞后的动态变化.方法 建立乳大鼠大脑皮层星形胶质细胞的体外培养体系,用HTNV 76-118和SEOV L99分别感染乳大鼠大脑皮层星形胶质细胞,免疫荧光染色、Western印迹和RT-PCR检测病毒感染细胞后病毒NP和S基因片段的变化.结果 成功建立了乳大鼠大脑皮层星形胶质细胞的体外培养体系,并通过免疫荧光染色、Western印迹和RT-PCR证实HTNV 76-118和SEOV L99可以感染离体培养的乳大鼠大脑皮层星形胶质细胞,随感染时间推移,病毒感染细胞数和病毒量显著增加.结论 乳大鼠大脑皮层星形胶质细胞可作为HTNV和SEOV感染的靶细胞.这为研究肾综合征出血热发病机制提供了实验平台.
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abstractsObjective To confirm that astrocytes from cerebral cortex of newborn rat can be the target cells of Hantaan virus (HTNV)and Seoul virus (SEOV)infection and to observe changes of astrocytes after different infection time. Methods Astrocytes were prepared from cerebral cortex of newborn rat, and then infected with HTNV and SEOV. The established virus infections were confirmed by detection of virus nucleocapsid protein (NP) and S segment RNA in astrocytes using double-label immunofluoreseence, Western-blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results The astrocytes from cerebral cortex of newborn rat were cultured successfully in vitro, which could be infected by HTNV and SEOV. The number of infected astrocytes and the virus titer in the infected astrocytes kept on increasing along with the extended infection duration. Conclusions Astrocytes from cerebral cortex of newborn rat are the target cells for HTNV and SEOV infection. Then establishment of in vitro cultured astrocytes model for Hantaviruses infection will be helpful for the study on the pathogenesis of hemorrhagic fever with renal syndrome.
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