Kurosaki M, Enomoto N, Marumo F, Sato C
Second Department of Internal Medicine, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Arch Virol. 1995;140(6):1087-94. doi: 10.1007/BF01315417.
In each infected patient, the population of hepatitis C virus is composed of quasispecies that differ in their nucleotide sequences. Among regions in hepatitis C virus genomes, nucleotide sequences of the hypervariable region have been shown to change quickly during the course of infection. It is not known, however, whether these variations exist in the core region that has recently been suggested to contain human lymphocyte antigen class 1 restricted sites for cytotoxic T cell recognition. To clarify this, RNA was extracted from the plasma of four patients with chronic hepatitis C. After cDNA synthesis, DNA fragments that contain the core region were amplified by the polymerase chain reaction and the diversity of the core region was analyzed by the single strand conformation polymorphism analysis. Using this method, single or multiple DNA bands were observed in each patient, and representative bands showed different nucleotide sequences. Comparison of single strand conformation polymorphism patterns revealed that the population of quasispecies changed during the course of chronic infection. These changes were more remarkable in patients with high serum alanine aminotransferase levels than those with low serum alanine aminotransferase levels. Thus, sequential variations exist in the core region of hepatitis C virus in same individuals, and the population of quasispecies as determined by the sequence of the core region changes during the course of infection, which might be related to cytopathic effects of hepatitis C virus.
在每一位受感染患者体内，丙型肝炎病毒群体由核苷酸序列不同的准种组成。在丙型肝炎病毒基因组的各个区域中，高变区的核苷酸序列在感染过程中变化迅速。然而，目前尚不清楚这些变异是否存在于最近被认为含有细胞毒性T细胞识别的人类淋巴细胞抗原1类限制性位点的核心区域。为了阐明这一点，从四名慢性丙型肝炎患者的血浆中提取了RNA。在合成cDNA后，通过聚合酶链反应扩增包含核心区域的DNA片段，并通过单链构象多态性分析来分析核心区域的多样性。使用这种方法，在每位患者中均观察到单条或多条DNA条带，且代表性条带显示出不同的核苷酸序列。单链构象多态性模式的比较显示，在慢性感染过程中准种群体发生了变化。血清丙氨酸氨基转移酶水平高的患者比血清丙氨酸氨基转移酶水平低的患者变化更显著。因此，同一患者体内丙型肝炎病毒的核心区域存在序列变异，并且由核心区域序列所确定的准种群体在感染过程中发生变化，这可能与丙型肝炎病毒的细胞病变效应有关。
