Ekman Anna, Hokynar Kati, Kakkola Laura, Kantola Kalle, Hedman Lea, Bondén Heidi, Gessner Matthias, Aberham Claudia, Norja Päivi, Miettinen Simo, Hedman Klaus, Söderlund-Venermo Maria
Department of Virology, Haartman Institute, University of Helsinki, Helsinki, Finland.
J Virol. 2007 Jul;81(13):6927-35. doi: 10.1128/JVI.02713-06. Epub 2007 Apr 4.
The human parvovirus B19 is now divided into three genotypes: type 1 (prototype), type 2 (A6- and LaLi-like), and type 3 (V9-like). In overall DNA sequence, the three virus types differ by approximately 10%. The most striking DNA dissimilarity, of >20%, is observed within the p6 promoter region. Because of the scarcity of data on the biological activities and pathogenetic potentials of virus types 2 and 3, we examined the functional characteristics of these virus types. We found the activities of the three p6 promoters to be of equal strength and to be most active in B19-permissive cells. Virus type 2 capsid protein VP2, alone or together with VP1, was expressed with the baculovirus system and was shown to assemble into icosahedral parvovirus-like particles, which were reactive in the hemagglutination assay. Furthermore, sera containing DNA of any of the three B19 types were shown to hemagglutinate. The infectivities of these sera were examined in two B19-permissive cell lines. Reverse transcription-PCR revealed synthesis of spliced B19 mRNAs, and immunofluorescence verified the production of NS and VP proteins in the infected cells. All three genotypes showed similar functional characteristics in all experiments performed, showing that the three virus types indeed belong to the same species, i.e., human parvovirus B19. Additionally, the antibody activity in sera from B19 type 1- or type 2-infected subjects (long-term immunity) was examined with homo- and heterologous virus-like particles. Cross-reactivity of 100% was observed, indicating that the two B19 genotypes comprise a single serotype.
人细小病毒B19现分为三种基因型:1型(原型)、2型(A6样和LaLi样)和3型(V9样)。在总体DNA序列上,这三种病毒类型相差约10%。在p6启动子区域观察到最显著的DNA差异,超过20%。由于关于2型和3型病毒的生物学活性和致病潜力的数据稀缺,我们研究了这些病毒类型的功能特性。我们发现这三种p6启动子的活性强度相同,且在对B19敏感的细胞中活性最强。2型病毒衣壳蛋白VP2单独或与VP1一起,通过杆状病毒系统表达,并显示组装成二十面体细小病毒样颗粒,这些颗粒在血凝试验中有反应。此外,含有三种B19类型中任何一种DNA的血清都显示有血凝活性。在两种对B19敏感的细胞系中检测了这些血清的感染性。逆转录聚合酶链反应显示合成了剪接的B19 mRNA,免疫荧光证实了感染细胞中NS和VP蛋白的产生。在所有进行的实验中,所有三种基因型都表现出相似的功能特性,表明这三种病毒类型确实属于同一物种,即人细小病毒B19。此外,用同源和异源病毒样颗粒检测了1型或2型B19感染受试者血清(长期免疫)中的抗体活性。观察到100%的交叉反应性,表明这两种B19基因型构成单一血清型。