丙型肝炎病毒对人B淋巴细胞系的感染及其形态学研究

Hepatitis C virus infection and morphologic study in B lymphocytes of patient with hepatitis C

目的 研究ＨＣＶ对人Ｂ淋巴细胞的感染，建立ＨＣＶ感染的丙型肝炎患者Ｂ淋巴细胞模型（ＣＢＣＬ），并进行ＨＣＶ的形态学研究。方法 用ＥＢ病毒转化Ｂ细胞建立Ｂ细胞系，以逆转录多聚酶链反应（ＲＴ－ＰＣＲ）、免疫组织化学、原位杂交方法检测Ｂ细胞系上清液及细胞内的ＨＣＶ抗原及ＨＣＶ ＲＮＡ，并通过电镜对ＨＣＶ进行形态学研究。结果 细胞系上清液中ＨＣＶ ＲＮＡ呈阳性，细胞内ＨＣＶ抗原及ＨＣＶ ＲＮＡ均呈阳性，电镜观察结果发现细胞内存在６５ｎｍ和１１０ｎｍ圆球型病毒颗粒，并可见病毒芽生形成现象。结论 ＨＣＶ可感染人Ｂ淋巴细胞并在其中复制。病毒在感染细胞胞质空泡部位合成和组装，以芽生方式进入胞质空泡内形成病毒颗粒。

Objective To study hepatitis C Virus (HCV) infection in B lymphocytes of patients with hepatitis C and to establish a B cell line with HCV infection and observe the hepatitis C virus like particles. Methods A B lymphoblastoid cell line was established by Epstein-Barr virus induced transformation directly from peripheral blood mononuclecyte cells of a patient with hepatitis C. The HCV antigen and HCV RNA were detected by immunohistochemical technique. Reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization and HCV particle were de tected by electron microscopy. Results Positive HCV RNA was found in supernatants of B cell line. HCV Ag and HCV RNA were also showed positive. Electron microscopy observed HCV spherical virus like particles with a diameter of approximately 65 um and 110 urn and the 'bud mutation' of HCV in the cytoplasmic vesicles of B lymphocytes. Conclusions HCV could infect B lymphocytes and replicate in the cell line. HCV particles are formed by 'bud mutation' of HCV in the cytoplasmic vesicles of B-lymphocytes.