Sepp T, Wang A L, Wang C C
Department of Pharmaceutical Chemistry, University of California, San Francisco 94143-0446.
J Virol. 1994 Mar;68(3):1426-31. doi: 10.1128/JVI.68.3.1426-1431.1994.
Giardia lamblia virus (GLV) is a small nonenveloped double-stranded RNA virus that infects specifically the parasitic protozoan G. lamblia. Among the many collected strains of G. lamblia, a few turn out to be highly resistant to the virus infection. Two of these strains, Ac and JH, were subjected to electroporation with the RNA from GLV-infected G. lamblia WB strain. Subsequent studies indicated the presence of GLV double-stranded RNA and GLV protein in the electroporated and propagated cells. Virus particles, released by the transfected cells into the culture medium, were capable of infecting the virus-sensitive G. lamblia WB strain. When the WB cells were incubated with GLV at 4 degrees C and treated with the bifunctional cross-linking reagent disuccinimidyl suberate, little GLV protein was detectable inside the cells by immunofluorescent staining. However, patches of fluorescent granules were found on the membrane surface of the cells, suggesting cross-linking of the viruses with a certain membrane component(s). Similar treatment of the resistant strains Ac and JH showed no fluorescence either inside or outside of the cells. Two other closely related parasitic protozoa, Tritrichomonas foetus and Trichomonas vaginalis, cannot be infected by GLV via either viral infection or RNA transfection. The [35S]cysteine-labeled protein profiles in Triton X-114 extracts of G. lamblia WB, Ac, and JH were compared. The profile of the WB strain differs clearly from that of Ac and JH. It remains to be seen, however, whether this difference is related at all to the different susceptibilities to GLV infection.
贾第虫病毒(GLV)是一种小型无包膜双链RNA病毒,专门感染寄生原生动物蓝氏贾第鞭毛虫。在众多收集的蓝氏贾第鞭毛虫菌株中,有少数对病毒感染具有高度抗性。其中两个菌株,即Ac和JH,用来自感染GLV的蓝氏贾第鞭毛虫WB菌株的RNA进行电穿孔处理。随后的研究表明,在电穿孔和传代培养的细胞中存在GLV双链RNA和GLV蛋白。转染细胞释放到培养基中的病毒颗粒能够感染对病毒敏感的蓝氏贾第鞭毛虫WB菌株。当WB细胞在4℃下与GLV孵育并用双功能交联剂辛二酸二琥珀酰亚胺酯处理时,通过免疫荧光染色在细胞内几乎检测不到GLV蛋白。然而,在细胞的膜表面发现了荧光颗粒斑块,表明病毒与某种膜成分发生了交联。对抗性菌株Ac和JH进行类似处理,细胞内外均未显示荧光。另外两种密切相关的寄生原生动物,胎儿三毛滴虫和阴道毛滴虫,无论是通过病毒感染还是RNA转染都不能被GLV感染。比较了蓝氏贾第鞭毛虫WB、Ac和JH的Triton X-114提取物中的[35S]半胱氨酸标记蛋白谱。WB菌株的图谱与Ac和JH的图谱明显不同。然而,这种差异是否与对GLV感染的不同易感性有关还有待观察。