Construction and characterization of a replication-defective herpes simplex virus 2 ICP8 mutant strain and its use in immunization studies in a guinea pig model of genital disease.
Construction and characterization of a replication-defective herpes simplex virus 2 ICP8 mutant strain and its use in immunization studies in a guinea pig model of genital disease.
复制标题
复制缺陷型单纯疱疹病毒 2 ICP8 突变株的构建和表征及其在豚鼠生殖器疾病模型免疫研究中的应用。
DOI：
10.1006/viro.1997.8564
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Knipe,DM
中科院分区：
文献类型：
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作者：
DaCosta,XJ;Bourne,N;Stanberry,LR;Knipe,DM
用单纯疱疹病毒1 (HSV-1) HD-2突变株的ICP8 - lacz融合基因取代单纯疱疹病毒2 (HSV-2) 186株的ICP8基因，构建了一种复制缺陷突变株。由此产生的病毒HSV-2 5BlacZ在Vero细胞中生长有缺陷，但能够在表达HSV-1 ICP8的细胞系中生长。在Vero细胞中，突变病毒在DNA合成方面存在缺陷，但能够以与野生型病毒相似的水平表达许多病毒蛋白，包括一些晚期动力学类病毒。SDS-PAGE和Western blot分析显示5BlacZ在Vero细胞中表达糖蛋白B和D。初步研究表明，5BlacZ免疫可保护豚鼠免受阴道内HSV-2的攻击。免疫动物的生殖器皮肤病较轻，初次感染时攻击病毒在生殖道的复制减少，疾病复发次数减少。因此，HSV-2 ICP8表现出与HSV-1 ICP8相似的基因调控特性，该HSV-2 ICP8突变病毒表现出与HSV-1 ICP8突变株相似的表型。2型单纯疱疹病毒复制缺陷突变体为免疫干预2型单纯疱疹病毒生殖器疾病和潜伏感染提供了一种潜在的疫苗途径。
A replication-defective mutant of herpes simplex virus 2 (HSV-2) was engineered by replacing the ICP8 gene of HSV-2 strain 186 with an ICP8–lacZ fusion gene from the herpes simplex virus 1 (HSV-1) HD-2 mutant strain. The resulting virus, HSV-2 5BlacZ, is defective for growth in Vero cells but is capable of growth in a cell line that expresses HSV-1 ICP8. In Vero cells, the mutant virus is defective for DNA synthesis but is able to express many viral proteins at levels similar to those of wild-type virus, including several of the late kinetic class. SDS–PAGE and Western blot analysis demonstrated the expression of glycoproteins B and D by 5BlacZ in Vero cells. Initial studies have shown that immunization with 5BlacZ protects guinea pigs from intravaginal HSV-2 challenge. Immunized animals had less severe genital skin disease and reduced replication of the challenge virus in the genital tract during primary infection and reduced episodes of recurrent disease. Thus, HSV-2 ICP8 shows gene regulatory properties similar to those of HSV-1 ICP8, and this HSV-2 ICP8 mutant virus shows a phenotype similar to those of HSV-1 ICP8 mutant strains. Replication-defective mutants of HSV-2 offer a potential vaccine approach for immune intervention against HSV-2 genital disease and latent infection.