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为了初步探索发热伴血小板减少综合征病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)核蛋白(nucleoprotein,NP)对宿主细胞免疫功能的影响。将SFTSV NP和NSs蛋白的编码基因插入真核表达载体VR1012中,通过免疫共沉淀(IP)、SDS-PAGE、质谱检测及蛋白质免疫印迹等方法寻找宿主细胞中与NP相互作用,同时又与免疫功能有关的蛋白质分子,并利用细胞免疫荧光方法检测SFTSV NP与该分子在细胞中的共定位情况。IP和质谱检测结果显示NP能与免疫功能相关的60kD SSA/Ro蛋白发生特异性结合,细胞免疫荧光试验进一步显示NP与60kD SSA/Ro蛋白在细胞质中存在共定位。提示SFTSV可能通过其核蛋白与免疫相关分子60kD SSA/Ro蛋白发生特异性结合,从而引起机体一系列的免疫反应和临床症状。
To investigate the effect of nucleoprotein (NP) on the immune function of host cells with Severe fever with thrombocytopenia syndrome virus (SFTSV). The gene encoding SFTSV NP and NSs was inserted into the eukaryotic expression vector VR1012 to find the host cell interaction with NP by immunoprecipitation (IP), SDS-PAGE, mass spectrometry and Western blotting, and at the same time, Function-related protein molecules, and the use of cellular immunofluorescence method SFTSV NP and the molecular co-localization of the cells. The results of IP and mass spectrometry showed that NP could specifically bind to 60kD SSA / Ro protein related to immune function, and the immunofluorescence assay further showed colocalization between NP and 60kD SSA / Ro protein in the cytoplasm. It is suggested that SFTSV may specifically bind to its immune-related molecule 60kD SSA / Ro protein through its nuclear protein, causing a series of immune responses and clinical symptoms in the body.