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本文报道在内质网和细胞浆内稳定表达抗汉坦病毒核蛋白细胞内抗体,研究抗汉坦病毒核蛋白细胞内抗体抗病毒作用及其抗病毒作用机理。在获得稳定表达抗汉坦病毒核蛋白细胞内抗体的细胞系的基础上,用MTT法检测细胞内表达抗汉坦病毒单链抗体对细胞增殖的影响,证实细胞内抗体的表达不会影响到细胞的生长。不同时间收集病毒感染毒后细胞培养培养上清和细胞裂解物,用ELISA方法检测细胞内外病毒结构蛋白含量的变化,结果表明定位于细胞内质肉和细胞浆内的抗汉坦病毒核蛋白细胞内抗体均能不同程度降低感染细胞上清中的核蛋白含量,但是不能或轻微抑制细胞内汉坦病毒核蛋白的合成。利用病毒微量滴定免疫荧光方法,对细胞内抗体与病毒的增殖关系进行了研究,证实无论在内质网还是在胞质,抗核蛋白细胞内抗体都能够抑制病毒的增殖,具有抗病毒活性。抗汉坦病毒核蛋白细胞内抗体不能抑制病毒结构蛋白的合成,其抗病毒效果是通过抑制病毒的包装而实现。
Abstract：In this study,we have investigated the potential functions of Hantavirus nuclecapsid(N) protein in Hantavirus replication,assembly and maturation by expression of the intracellular antibodies to Hantavirus nucleocapsid(N) protein localized in endoplasmic reticulum(ER) and cytoplasma(Cyto) of Vero-E6 cells.The stable cell lines expressing intracellular antibodies to N protein were constructed,the cell proliferations between each intracellular antibody expression cell line and normal Vero-E6 cell line were compared with the method of MTT,and showed no significant difference between above two cell lines.Further,virus-infected cell culture supernatants and cell lysates were collected and tested at different infection times. The N protein concentration in the supernatants of virus-infected cells were reduced at different levels in all ER and cytoplasma localized intracellular antibody expressing cell lines,however,the intracellular N protein synthesis in above cell lines was not significantly affected.By using microtitration test with immunofluorescence assay,the effect of intracellular antibodies on the multiplication of Hantavirus in infected cells were investigated,and the results showed significant inhibition of viral multiplication with both ER and Cyto localized antibodies.The antiviral efficiency of the intracellular antibodies to N protein may take mechanisms of inhibiting virus assembly but not the viral protein synthesis.
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基本信息:
中图分类号:R392
引用信息:
[1]王涛,李建东,张全福,等.抗汉坦病毒核蛋白细胞内抗体抗病毒作用的研究[J].病毒学报,2006(02):89-95.
基金信息:
国家自然科学基金项目(编号:30371269)
2006-03-30
2006-03-30