EXPRESSION OF HEPATITIS C VIRUS ENVELOPE RELATED PROTEINS AND DETECTION OF ANTIBODIES TO PROTEINS.

丙型肝炎病毒包膜相关蛋白的表达和蛋白抗体的检测。

• 批准号: 04670408
• 负责人: YOKOSUKA Osamu
• 金额: $ 1.34万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670408/
• 关键词: Hepatitis C virus; HCV-RNA; Neutralizing antibody; E2 antibody; Expression vector; E2抗体; C型肝炎ウイルス; HCV表面関連抗原(EZ); 表面関連抗原(E2)

hepatitis C virus (HCV) is a major causative agent of ahronic liver diseases in Japan. M.Houghtom et al. reported a partial sequence of HCV and detection systems of anti-HCV nonstructural proteins and core proteins were developed. We tried to establish a system to detect antibodies to envelope related proteins of HCV and examined the clinical meanig of the antigen.antibody system.We cloned the genes coding the HCV membrace related proteins (E2/NS1) from sera of patients with chronic type C hapatitis. Then, we inserted the genes to trotein expression vector and expressed these E2/NS1 proteins in E.Coli and purified. After SDS gel electrophoresis, the E2 proteins were transferd to nitro cellulose membrane and western blotting was performed with sera from various patients including type C hepatitis. We further improved the detection method by not strongly denaturing the E2 proteins, and as a results higher detection rate of the antibody was obtained. We also examined the cases who were treated with interferon and found the anti-E2 antibody decreased after the successful reeatment. From these results, it seems anti-E2 antibody is not a efficient neutralizing antibody incontrast to other flaviviruses, although it is possible that the antibody may from immune-complex with HCV.

丙型肝炎病毒（HCV）是日本慢性肝病的主要病原体。 M.Houghtom 等人。报道了HCV的部分序列，并开发了抗HCV非结构蛋白和核心蛋白的检测系统。我们尝试建立HCV包膜相关蛋白抗体检测系统，并探讨该抗原抗体系统的临床意义。我们从慢性丙型肝炎患者血清中克隆了编码HCV包膜相关蛋白（E2/NS1）的基因。然后，我们将基因插入到蛋白表达载体中，并在大肠杆菌中表达这些E2/NS1蛋白并纯化。 SDS凝胶电泳后，E2蛋白被转移到硝酸纤维素膜上，并用来自包括丙型肝炎在内的各种患者的血清进行蛋白质印迹分析。我们进一步改进了检测方法，不对E2蛋白进行强烈变性，结果获得了更高的抗体检出率。我们还检查了接受干扰素治疗的病例，发现治疗成功后抗E2抗体有所下降。从这些结果来看，与其他黄病毒相比，抗E2抗体似乎不是有效的中和抗体，尽管该抗体可能来自与HCV的免疫复合物。

项目成果

Yokosuka Osamu: "Expression of HCV E2/NS1 protein as a fusion protein with maltose binding protein." Gut. 34. S64-S65 (1993)

Yokosuka Osamu：“HCV E2/NS1 蛋白作为与麦芽糖结合蛋白的融合蛋白的表达。”

Yokosuka O, Ito Y, Imazeki F, Ohto M, Omata M: "Detection of antibody to hepatitis C E2/NS1 protein in phtients with type C hepatitis" Biochem Biophy Res commun. 189. 565-571 (1992)

Yokosuka O、Ito Y、Imazeki F、Ohto M、Omata M：“丙型肝炎患者中丙型肝炎 E2/NS1 蛋白抗体的检测”Biochem Biophy Res commun。

Yokosuka O, Ito Y, Sakuna J, Imazeki F, Ohto M, Omata M: "Expression of hepatitis C virus core protein as a fusion protein with maltose binding protein" Dig Dis Sci. 38. 626-630 (1993)

Yokosuka O、Ito Y、Sakuna J、Imazeki F、Ohto M、Omata M：“丙型肝炎病毒核心蛋白作为与麦芽糖结合蛋白的融合蛋白的表达”Dig Dis Sci。

Yokosuka O, Omata M, Ito Y, Ohto M: "Expression of HCV E2/NS1 protein as a fusion protein with maltose binding protein." Gut. 34. S64-S65 (1993)

Yokosuka O、Omata M、Ito Y、Ohto M：“HCV E2/NS1 蛋白作为与麦芽糖结合蛋白的融合蛋白的表达。”

Yokosuka Osamu: "Expression of hepatitis C virus core protein as a fusion protein with malfose binding protein." Digestive Disease and Science. 38. 626-630 (1993)

Yokosuka Osamu：“丙型肝炎病毒核心蛋白作为与马尔福糖结合蛋白的融合蛋白的表达。”