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目的 探讨汉滩病毒 (HV)感染诱导乳鼠组织热休克蛋白 (HSPs)表达及热休克蛋白与病毒蛋白的相互关系。方法 出生后 2～ 3d的昆明乳鼠 ,每只脑内接种 10 0个半数致死量的HV0 .0 3ml,8d后乳鼠发病死亡 ,取其脑组织一部分固定 ,石蜡包埋切片 ,用免疫组织化学法结合共聚焦显微镜检测组织中病毒抗原及HSP70的表达 ,一部分组织制匀浆液 ,用ELISA、免疫共沉淀方法分析病毒抗原和HSP70的表达及关系。结果 HV感染诱导了乳鼠脑组织神经细胞高表达热休克蛋白(HSP) 70 ,细胞内病毒抗原与HSP70有共定位关系 ;免疫共沉淀结果显示用HSP70mAb可以共沉淀感染乳鼠脑组织匀浆中的汉滩病毒核心抗原 (HV NP) ,用HV NPmAb 1A8可沉淀感染乳鼠脑组织匀浆中的HSP70 ,对照实验阴性 ,所用两种蛋白抗体间无交叉反应 ,因而HSP70与HV NP结合形成复合物 ,结果是特异的。结论 HV感染诱导HSP70的表达 ,后者与HV NP结合形成HSP70 病毒抗原肽复合物 ,该复合物可能在病毒感染和复制过程中发挥作用。
Objective To investigate the expression of heat shock protein (HSPs) and the relationship between heat shock protein (HSP) and viral proteins induced by Hantaan virus (HV) infection in neonatal rats. Methods Kunming neonatal rats 2 ~ 3d after birth were vaccinated with 100 ml of HV0.030 in half a lethal dose in each brain. After 8 days, the sufferers were killed and their brains were partially fixed. Paraffin-embedded sections were stained with immunohistochemistry Chemotherapy and confocal microscopy were used to detect the expression of viral antigens and HSP70 in the tissues. Some tissues were homogenized and the expression of the viral antigens and HSP70 were analyzed by ELISA and co-immunoprecipitation. Results HV infection induced high expression of heat shock protein (HSP70) in neural cells of neonatal rat brain tissue. Intracellular viral antigens co-localized with HSP70. Co-immunoprecipitation showed HSP70 mAb could be co-precipitated in homogenized neonatal rat brain homogenates Of Hantaan virus core antigen (HV NP). HV NPmAb 1A8 precipitated HSP70 from infected neonatal rat brain homogenates. The control experiment was negative. There was no cross-reaction between the two protein antibodies used, so HSP70 and HV NP bound to form complex The result is specific. Conclusion HV infection induces the expression of HSP70, which binds to HV NP to form HSP70 antigen complex, which may play a role in virus infection and replication.