ANTIGENIC AND GENETIC-VARIATION IN CYTOPATHIC HEPATITIS-A VIRUS VARIANTS ARISING DURING PERSISTENT INFECTION - EVIDENCE FOR GENETIC-RECOMBINATION
ANTIGENIC AND GENETIC-VARIATION IN CYTOPATHIC HEPATITIS-A VIRUS VARIANTS ARISING DURING PERSISTENT INFECTION - EVIDENCE FOR GENETIC-RECOMBINATION
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DOI：
10.1128/jvi.65.4.2056-2065.1991
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发表时间：
1991-04-01
影响因子：
5.4
通讯作者：
JANSEN, RW
中科院分区：
文献类型：
--
作者：
LEMON, SM;MURPHY, PC;JANSEN, RW
从持续感染的绿色猴肾（BS-C-1）细胞中回收的甲型肝炎病毒（pHM 175病毒）变体在BS-C-1或胎猴肾（FRhK-4）细胞中连续传代期间诱导细胞病变效应。 表位特异性放射免疫聚焦试验表明，该病毒包括两个病毒粒子群体，一个具有改变的抗原性，包括对单克隆抗体K24 F2的中和抗性，另一个具有正常的抗原特性。 在持续感染期间，抗原变体的复制比具有正常抗原表型的病毒更有利，而在连续传代期间选择具有正常抗原表型的病毒。 每种类型的病毒进行克隆分离，都是细胞病变的细胞培养物，并显示快速复制表型相比，非细胞病变的第16代（p16）HM 175病毒，用于建立原始的持续感染。 这两个细胞病变病毒克隆与p16 HM 175序列相比分别含有31和34个核苷酸的变化。 两者共享共同的5'序列（碱基30至1677），以及P2-P3区（碱基3249至5303和6462至6781）和3'末端（碱基7272至7478）中的序列同一性。 VP 3、VP 1和3C（pro）在两个病毒克隆中含有不同的突变，在抗原变体的VP 3的第70位残基和VP 1的第197和276位残基处具有氨基酸取代。 这些衣壳突变不影响病毒粒子的热稳定性。 对三种克隆分离的致细胞病变变异体的几乎完整的基因组序列进行比较，提示这些病毒在持续感染期间发生了遗传重组，并表明5'和3'非翻译区以及非结构蛋白2A、2B、2C、3A和3D（pol）的突变可能与致细胞病变表型有关。
Variants of hepatitis A virus (pHM175 virus) recovered from persistently infected green monkey kidney (BS-C-1) cells induced a cytopathic effect during serial passage in BS-C-1 or fetal rhesus kidney (FRhK-4) cells. Epitope-specific radioimmunofocus assays showed that this virus comprised two virion populations, one with altered antigenicity including neutralization resistance to monoclonal antibody K24F2, and the other with normal antigenic characteristics. Replication of the antigenic variant was favored over that of virus with the normal antigenic phenotype during persistent infection, while virus with the normal antigenic phenotype was selected during serial passage. Viruses of each type were clonally isolated; both wer cytopathic in cell cultures and displayed a rapid replication phenotype when compared with the noncytopathic passage 16 (p16) HM175 virus which was used to establish the original persistent infection. The two cytopathic virus clones contained 31 and 34 nucleotide changes from the sequence of p16 HM175. Both shared a common 5' sequence (bases 30 to 1677), as well as sequence identity in the P2-P3 region (bases 3249 to 5303 and 6462 to 6781) and 3' terminus (bases 7272 to 7478). VP3, VP1, and 3C(pro) contained different mutations in the two virus clones, with amino acid substitutions at residues 70 of VP3 and 197 and 276 of VP1 of the antigenic variant. These capsid mutations did not affect virion thermal stability. A comparison of the nearly complete genomic sequences of three clonally isolated cytopathic variants was suggestive of genetic recombination between these viruses during persistent infection and indicated that mutations in both 5' and 3' nontranslated regions and in the nonstructural proteins 2A, 2B, 2C, 3A, and 3D(pol) may be related to the cytopathic phenotype.