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目的:寻找Vero细胞膜上可与JEV结合的分子。方法:采用免疫共沉淀(Co-IP)技术,分离出Vero细胞膜上可与日本脑炎病毒(JEV)结合的分子,做质谱(MS)分析针对特异结合分子进行检测。用流式细胞术(FCM)和免疫荧光实验(IFA)验证候选结合分子与JEV结合的情况。结果:Co-IP产物经SDS-PAGE分离出3条可以与JEV分子特异性结合的条带。其中,MS分析只鉴定出相对分子质量(Mr)约为90000的分子为热休克蛋白90β(HSP90β)。FCM及IFA实验结果表明抗HSP90β单克隆抗体(mAb)可以明显阻断JEV与Vero细胞结合。结论:Vero细胞膜上的HSP90β能与JEV结合。
OBJECTIVE: To find molecules that bind to JEV in Vero cell membrane. METHODS: Co-IP technique was used to separate the molecules that could bind to Japanese encephalitis virus (JEV) on the Vero cell membrane. MS detection was performed on specific binding molecules by mass spectrometry (MS). Flow cytometry (FCM) and immunofluorescence experiments (IFA) were used to verify the binding of candidate binding molecules to JEV. Results: Three Co-IP products were separated by SDS-PAGE. Among them, MS analysis identified only the molecule with a relative molecular mass (Mr) of about 90,000 as heat shock protein 90β (HSP90β). FCM and IFA results show that anti-HSP90β monoclonal antibody (mAb) can significantly block JEV and Vero cells. Conclusion: HSP90β in Vero cell membrane can bind with JEV.