THE VIRUS OF HEPATITIS B

乙型肝炎病毒

• 批准号: 3480748
• 负责人: WILLIAM S ROBINSON
• 金额: $ 40.17万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-06-01 至 1995-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3480748
• 关键词: Rotavirus; carcinoma; chimeric proteins; duck hepatitis B virus; electron microscopy; genetic manipulation; hepatitis B antigens; hepatitis vaccine; hepatocellular carcinoma; human subject; immunogenetics; laboratory rabbit; laboratory rat; liver regeneration; microorganism immunology; monoclonal antibody; passive immunization; proteins; squirrel; surface antigens; tissue /cell culture; virion; virus DNA; virus cytopathogenic effect; virus genetics; virus replication

Hepatitis B virus (HBV) is an important cause of liver disease including liver cancer in man. It is a member of a family of viruses which have interesting structure, mode of replication and biological properties. We propose to study the role of HBV in liver cancer by analyzing integrated viral DNA sequences with flanking cellular DNA cloned from two hepatocellular carcinomas (HCC), each with single integration sites. We will also further investigate NIH 3T3 cell transformation observed with a cloned HBV DNA dimer, an 1850 bp BamHI fragment of HBV DNA and DNA from HCC. We will also investigate the effect of liver regeneration on viral integration and HCC formation. We will further characterize high molecular weight HBV DNA forms (5 and 9 kd) found in virions from plasma. A final goal is to investigate a viral vaccine strategy by which relevant viral antigens are expressed by enteric bacterial strains which result in immune responses but not disease when live bacteria are ingested by mouth. Although this approach might offer a particular advantage for enteric pathogens such as rotavirus for which local enteric immunity is important, the approach could be equally effective for other agents such as HBV for which only systemic immunity is important. We have introduced genes for hepatitis B surface (HBsAg) and core (HBcAg) antigens and rotavirus VP7 into plasmid expression vectors and have achieved expression of HBsAg and rotavirus VP7 as fusion proteins and HBcAg as a discrete polypeptide in E. coli. We propose to optimize expression of these antigens in E. coli; introduce the hybrid plasmids into Salmonella typhi strain Ty 21a and S. typhimurine 1479 (mutants with attenuated virulence and successfully used as live, oral Salmonella vaccines); and test the transformed Salmonella for viral antigen expression and ability to induce immune responses in mice. The long-term goal is live oral vaccines for rotavirus and HBV in man.

乙型肝炎病毒（HBV）是导致肝脏疾病的重要原因，包括 男性肝癌。 它是病毒家族的一员，具有 有趣的结构、复制模式和生物学特性。 我们 提议通过综合分析研究 HBV 在肝癌中的作用 病毒 DNA 序列与从两个细胞克隆的侧翼细胞 DNA 肝细胞癌（HCC），每种都有单一的整合位点。 我们 还将进一步研究 NIH 3T3 细胞转化观察到的 克隆的 HBV DNA 二聚体，HBV DNA 的 1850 bp BamHI 片段和来自 肝癌。 我们还将研究肝再生对病毒的影响 整合和 HCC 形成。 我们将进一步表征高分子 血浆病毒粒子中发现的 HBV DNA 形式（5 和 9 kd）的重量。 决赛 目标是研究一种病毒疫苗策略，通过该策略，相关病毒 抗原由肠道细菌菌株表达，从而产生免疫 当活细菌被口腔摄入时，会产生反应，但不会导致疾病。 尽管这种方法可能为肠溶性药物提供了特殊的优势 病原体，例如轮状病毒，局部肠道免疫对其很重要， 该方法对于其他药物（例如 HBV）同样有效 只有系统免疫才是重要的。 我们引入了基因 乙型肝炎表面 (HBsAg) 和核心 (HBcAg) 抗原以及轮状病毒 VP7 转入质粒表达载体并实现了 HBsAg 和 轮状病毒 VP7 作为融合蛋白，HBcAg 作为大肠杆菌中的离散多肽。 大肠杆菌。 我们建议优化这些抗原在大肠杆菌中的表达； 将杂交质粒引入伤寒沙门氏菌菌株 Ty 21a 和 S。 伤寒鼠碱 1479（毒力减弱的突变体，已成功用于 如口服沙门氏菌活疫苗）；并测试转化的沙门氏菌 病毒抗原表达和诱导小鼠免疫反应的能力。 长期目标是为人类提供针对轮状病毒和乙型肝炎病毒的口服活疫苗。