Analysis of Hepatitis B Virus Expression, Using a Helper-Free Adenovirus Vector.

Research Project

Project/Area Number	61570231
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	Virology
Research Institution	The National Institute of Health of Japan
Principal Investigator	YUASA Tazuko NIH of Japan: Enteroviruses Unit, 腸内ウイルス部, 主任研究官 (80100105)
Co-Investigator(Kenkyū-buntansha)	MIYAMURA Tatsuo NIH of Japan: Enteroviruses Unit, 腸内ウイルス部, 室長 (90100099)
Project Period (FY)	1986 – 1988
Project Status	Completed (Fiscal Year 1988)
Budget Amount *help	¥2,400,000 (Direct Cost: ¥2,400,000) Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 1986: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywords	Hepatitis B virus / Adenovirus / HB surface antigen (HBsAg) / Large HBsAg / 小型HBs抗原 / アデノウイルスベクター
Research Abstract	Hepatitis B virus (HBV) has a very narrow host range and replicates only in human and chimpamzee liver. The in vitro replications of HBV have been also restricted to the human cell lines from the liver. The narrow host range of HBV might depend upon the tissue-or species-specificity of the HBV gene expressions. A recombinant adenovirus type 5 bearing 87% of HBV genome (Ad5-HBL) had been constructed and several HBV mRNAs transcribed from this recombinant virus in human HeLa cells were reported (Saito et al., 1985). Ad5-HBL has a wide host range, replicates to high copies per cell and HBV mRNAs are efficiently transcribed from its own promoters. So we infected Ad5-HBL to twelve cell lines of various species and tissue origins, and expression of HBV surface antigen (HBsAg) was examined by Sl mapping. We report here that 2.4Kb mRNA which codes large HBsAg is expressed abundantly in all human and monkey cells tested but 2.0Kb mRNA which codes middle and major HBsAg is expressed in rodent cells, irrespective of their original tissues. The ratios of the steady-state level of 2.4Kb mRNA coding for large HBsAg to that of 2.0Kb mrna coding for middle and major HBsAg are diffrent among cell lines, and show a species-specificity. By the radioimmunoassay and the analysis of SDS-polyacrylamide gel electrophoresis, large HBsAg was detected abundantly in human and monkey cells but major HBsAg was detected abundantly in rodent cells.