Parker J S, Parrish C R
James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
J Virol. 1997 Dec;71(12):9214-22. doi: 10.1128/JVI.71.12.9214-9222.1997.
We analyzed a region of the capsid of canine parvovirus (CPV) which determines the ability of the virus to infect canine cells. This region is distinct from those previously shown to determine the canine host range differences between CPV and feline panleukopenia virus. It lies on a ridge of the threefold spike of the capsid and is comprised of five interacting loops from three capsid protein monomers. We analyzed 12 mutants of CPV which contained amino acid changes in two adjacent loops exposed on the surface of this region. Nine mutants infected and grew in feline cells but were restricted in replication in one or the other of two canine cell lines tested. Three other mutants whose genomes contain mutations which affect one probable interchain bond were nonviable and could not be propagated in either canine or feline cells, although the VP1 and VP2 proteins from those mutants produced empty capsids when expressed from a plasmid vector. Although wild-type and mutant capsids bound to canine and feline cells in similar amounts, infection or viral DNA replication was greatly reduced after inoculation of canine cells with most of the mutants. The viral genomes of two host range-restricted mutants and two nonviable mutants replicated to wild-type levels in both feline and canine cells upon transfection with plasmid clones. The capsids of wild-type CPV and two mutants were similar in susceptibility to heat inactivation, but one of those mutants and one other were more stable against urea denaturation. Most mutations in this structural region altered the ability of monoclonal antibodies to recognize epitopes within a major neutralizing antigenic site, and that site could be subdivided into a number of distinct epitopes. These results argue that a specific structure of this region is required for CPV to retain its canine host range.
我们分析了犬细小病毒(CPV)衣壳的一个区域,该区域决定了病毒感染犬类细胞的能力。这个区域与先前显示的决定CPV和猫泛白细胞减少症病毒之间犬类宿主范围差异的区域不同。它位于衣壳三聚体刺突的一个脊上,由来自三个衣壳蛋白单体的五个相互作用的环组成。我们分析了12个CPV突变体,这些突变体在该区域表面暴露的两个相邻环中含有氨基酸变化。9个突变体在猫细胞中感染并生长,但在测试的两种犬类细胞系中的一种或另一种中复制受到限制。另外三个突变体,其基因组包含影响一个可能的链间键的突变,是无活力的,不能在犬类或猫类细胞中繁殖,尽管来自这些突变体的VP1和VP2蛋白在从质粒载体表达时产生空衣壳。尽管野生型和突变型衣壳以相似的量与犬类和猫类细胞结合,但用大多数突变体接种犬类细胞后,感染或病毒DNA复制大大减少。两个宿主范围受限的突变体和两个无活力突变体的病毒基因组在用质粒克隆转染后,在猫类和犬类细胞中都复制到野生型水平。野生型CPV和两个突变体的衣壳对热灭活的敏感性相似,但其中一个突变体和另一个对尿素变性更稳定。这个结构区域的大多数突变改变了单克隆抗体识别主要中和抗原位点内表位的能力,并且该位点可以细分为许多不同的表位。这些结果表明,该区域的特定结构是CPV保留其犬类宿主范围所必需的。