Fox J M, McCrackin Stevenson M A, Bloom M E
Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840, USA.
J Virol. 1999 Oct;73(10):8713-9. doi: 10.1128/JVI.73.10.8713-8719.1999.
Aleutian mink disease parvovirus (ADV) is the etiological agent of Aleutian disease of mink. Several ADV isolates have been identified which vary in the severity of the disease they elicit. The isolate ADV-Utah replicates to high levels in mink, causing severe Aleutian disease that results in death within 6 to 8 weeks, but does not replicate in Crandell feline kidney (CrFK) cells. In contrast, ADV-G replicates in CrFK cells but does not replicate in mink. The ability of the virus to replicate in vivo is determined by virally encoded determinants contained within a defined region of the VP2 gene (M. E. Bloom, J. M. Fox, B. D. Berry, K. L. Oie, and J. B. Wolfinbarger. Virology 251:288-296, 1998). Within this region, ADV-G and ADV-Utah differ at only five amino acid residues. To determine which of these five amino acid residues comprise the in vivo replication determinant, site-directed mutagenesis was performed to individually convert the amino acid residues of ADV-G to those of ADV-Utah. A virus in which the ADV-G VP2 residue at 534, histidine (H), was converted to an aspartic acid (D) of ADV-Utah replicated in CrFK cells as efficiently as ADV-G. H534D also replicated in mink, causing transient viremia at 30 days postinfection and a strong antibody response. Animals infected with this virus developed diffuse hepatocellular microvesicular steatosis, an abnormal accumulation of intracellular fat, but did not develop classical Aleutian disease. Thus, the substitution of an aspartic acid at residue 534 for a histidine allowed replication of ADV-G in mink, but the ability to replicate was not sufficient to cause classical Aleutian disease.
阿留申水貂病细小病毒(ADV)是水貂阿留申病的病原体。已鉴定出几种ADV分离株,它们引发疾病的严重程度各不相同。ADV-犹他分离株在水貂体内大量复制,导致严重的阿留申病,在6至8周内导致死亡,但在克兰德尔猫肾(CrFK)细胞中不复制。相比之下,ADV-G在CrFK细胞中复制,但在水貂中不复制。病毒在体内复制的能力由VP2基因特定区域内的病毒编码决定因素决定(M.E.布鲁姆、J.M.福克斯、B.D.贝里、K.L.奥伊和J.B.沃尔芬巴格。《病毒学》251:288-296,1998)。在该区域内,ADV-G和ADV-犹他仅在五个氨基酸残基上有所不同。为了确定这五个氨基酸残基中的哪一个构成体内复制决定因素,进行了定点诱变,将ADV-G的氨基酸残基逐个转化为ADV-犹他的氨基酸残基。一种病毒,其中ADV-G VP2在534位的组氨酸(H)被转化为ADV-犹他的天冬氨酸(D),在CrFK细胞中的复制效率与ADV-G一样高。H534D在水貂中也能复制,在感染后30天引起短暂病毒血症和强烈的抗体反应。感染这种病毒的动物出现弥漫性肝细胞微泡性脂肪变性,即细胞内脂肪的异常积累,但未发展为典型的阿留申病。因此,将534位的组氨酸替换为天冬氨酸可使ADV-G在水貂中复制,但复制能力不足以引起典型的阿留申病。