CHARACTERIZATION AND SEQUENCE ANALYSES OF ANTIBODY-SELECTED ANTIGENIC VARIANTS OF HERPES-SIMPLEX VIRUS SHOW A CONFORMATIONALLY COMPLEX EPITOPE ON GLYCOPROTEIN-H
CHARACTERIZATION AND SEQUENCE ANALYSES OF ANTIBODY-SELECTED ANTIGENIC VARIANTS OF HERPES-SIMPLEX VIRUS SHOW A CONFORMATIONALLY COMPLEX EPITOPE ON GLYCOPROTEIN-H
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DOI：
10.1128/jvi.65.5.2393-2401.1991
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发表时间：
1991-05-01
影响因子：
5.4
通讯作者：
MINSON, AC
中科院分区：
文献类型：
--
作者：
GOMPELS, UA;CARSS, AL;MINSON, AC
分离并鉴定了13株抗52S或LP11单抗中和的单纯疱疹病毒抗原变异株。在缺乏补体的情况下，抗体能有效地中和野生型病毒的感染性，并抑制病毒从感染细胞向未感染细胞的转移(“空斑抑制”)，并减少合胞体株对病毒诱导的细胞融合。第一个分离到的变种出现在体内。从100株临床分离株的分型研究中分析了66株1型分离株，其中1株被鉴定为抵抗LP11抗体的中和。获得了糖蛋白H(Gh)的序列，并与野生型和合胞体实验室菌株SC16、菌株17和Hfem的序列进行了比较。与观察到的不同亚群疱疹病毒Gh序列之间的多样性相比，这些序列高度保守。在任何一个比较中只有四个编码变化，并且在实验室菌株和临床分离株(Asp-168到Gly)之间只观察到一个独特的编码变化。将这些序列与组织培养中通过抗体选择的抗原变异体的序列进行比较。用亲本菌株SC16或HFEM中的抗体LP11或52S分别筛选出12个突变体。对于每个变异体，推导出Gh核苷酸序列，并鉴定出导致单一氨基酸替换的点突变。抗LP11的突变型病毒编码的Gh序列在Gh外区Glu-86、Asp-168或Arg-329的一半处有氨基酸替换，而抗52s的突变型病毒在邻近的Ser-536和Ala-537位置有替换。一株LP11突变病毒的Gh基因点突变与临床分离株相同，导致Asp-168被Gly取代。LP11和52S均识别不同的Gh表位为耐中和突变病毒，LP11免疫沉淀对52S敏感，而抗52S突变病毒则相反。这与以前的研究一致，这些研究表明，虽然52S表位可以在没有其他病毒产物的情况下形成，但LP11表位的稳定呈现和Gh到细胞表面的运输需要病毒基因的表达(Gompels和Minson，J.Virol)。63：4744-4755,1989)。除了一个变种病毒产量较低外，所有突变病毒产生的感染性颗粒数量都与野生型病毒相似。在野生型Gh产生细胞系上生长或滴定可以部分补充LP11突变病毒的抗体抗性表型，但突变株在抗菌斑抑制或中和感染性方面有所不同。综上所述，这些结果表明Gh具有一个构象复杂的表位，该表位是在病毒感染性和传播中具有重要功能的区域的一部分。当Gh在没有病毒感染的情况下表达时，这个表位是不稳定的，并且这个表位可能以不同的方式呈现在受感染的细胞表面或病毒被膜上。
Thirteen antigenic variants of herpes simplex virus which were resistant to neutralization by monoclonal antibody 52S or LP11 were isolated and characterized. The antibodies in the absence of complement potently neutralize infectivity of wild-type virus as well as inhibit the transfer of virus from infected to uninfected cells ("plaque inhibition") and decrease virus-induced cell fusion by syncytial strains. The first variant isolated arose in vivo. Of 66 type 1 isolates analyzed from typing studies of 100 clinical isolates, one was identified as resistant to neutralization by LP11 antibody. The glycoprotein H (gH) sequence was derived and compared with those of wild-type and syncytial laboratory strains SC16, strain 17, and HFEM. The sequences were highly conserved in contrast to the diversity observed between gH sequences from herpesviruses of different subgroups. Only four coding changes were present in any of the comparisons, and only one unique coding change was observed between the laboratory strains and the clinical isolate (Asp-168 to Gly). These sequences were compared with those of antigenic variants selected by antibody in tissue culture. Twelve variants were independently selected with antibody LP11 or 52S from parent strain SC16 or HFEM. For each variant, the gH nucleotide sequence was derived and a point mutation was identified giving rise to a single amino acid substitution. The LP11-resistant viruses encoded gH sequences with amino acid substitutions at sites distributed over one-half of the gH external domain, Glu-86, Asp-168, or Arg-329, while the 52S-resistant mutant viruses had substitutions at adjacent positions Ser-536 and Ala-537. One LP11 mutant virus had a point mutation in the gH gene that was identical to that of the clinical isolate, giving rise to a substitution of Asp-168 with Gly. Both LP11 and 52S appeared to recognize distinct gH epitopes as mutant virus resistant to neutralization and immunoprecipitation with LP11 remained sensitive to 52S and the converse was shown for the 52S-resistant mutant virus. This is consistent with previous studies which showed that while the 52S epitope could be formed in the absence of other virus products, virus gene expression was required for stable presentation of the LP11 epitope, and for transport of gH to the cell surface (Gompels and Minson, J. Virol. 63:4744-4755, 1989). All mutant viruses produced numbers of infectious particles that were similar to those produced by the wild-type virus, with the exception of one variant which produced lower yields. The antibody-resistant phenotype of the LP11 mutant viruses could be partially complemented by growth or titration on wild-type gH-producing cell lines, but the mutants differed in decreased resistance to plaque inhibition or neutralization of infectivity. Taken together, these results show that gH has a conformationally complex epitope which is part of a domain which has an essential function in virus infectivity and spread. This epitope is not stable when gH is expressed in the absence of virus infection, and this epitope may be differentially presented at the infected cell surface or on the viral envelope.