Abstract
We have demonstrated by immunoelectron microscopy that 42‐nm particles with double‐shelled structures characteristic of Dane particles are present in the serum of transgenic mice, 1.2HB‐BS 10, carrying partly duplicated hepatitis B virus (HBV) genome. Furthermore, these particles were shown to infect primary human fetal hepatocytes as demonstrated by the elevation of MBV surface antigen (HBsAg) in the culture medium. HBV DNA is known to be expressed in a liver‐ and kidney‐specific manner in the adult mouse, so we examined the developmental expression of viral antigens. In the liver, viral antigens (HBsAg and HBV e antigen) began to be expressed before birth and the level of expression showed a sharp rise after birth. On the other hand, in the kidney, viral antigens began to be expressed after birth. Serum levels of viral antigens were roughly proportional to the levels of expression in the liver, suggesting that the liver is the main source for viral antigens in the serum. None of these transgenic mice produced anti‐HBs or anti‐HBV core response or showed biochemical or pathological change up to at least 24 months of age. All these results suggest that infectious viral particles can be produced in transgenic mice, and that expression and replication of HBV DNA are not toxic in vivo.
Keywords: Transgenic mouse, Dane particle, Developmental expression
Full Text
The Full Text of this article is available as a PDF (299.1 KB).
REFERENCES
- 1. ) Beasly , R. P.Hepatitis B virus: the major etiology of hepatocellular carcinoma . Cancer , 61 , 1942 – 1956 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Mason , W. S. and Taylor , J.M. Experimental systems for the study of hepadnavirus and hepatitis delta virus infections . Hepatology , 9 , 635 – 645 ( 1989. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Tiollais , P. , Pourcel , C. and Dejean , A.The hepatitis B virus . Nature , 317 , 489 – 495 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 4. ) Araki , K. , Miyazaki , J. , Hino , O. , Tomita , N. , Chisaka , O. , Matsubara , K. and Yamamura , K.Expression and replication of hepatitis B virus genome in transgenic mice . Proc. Natl. Acad. Sci. USA , 86 , 207 – 211 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5. ) Tuttleman , J. S. , Pugh , J. C. and Summers , J. W.In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus . J. Virol , 58 , 17 – 25 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6. ) Sells , M. A. , Chen , M. L. and Acs , G.Production of hepatitis B virus particle in Hep G2 cells transfected with cloned hepatitis B virus DNA . Proc. Natl. Acad. Sci. USA , 84 , 1005 – 1009 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. ) Sureau , C. , Romet‐Lemonne , J. L. , Mullins , J. I. and Essex , M.Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA . Cell , 47 , 37 – 47 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 8. ) Tsurimoto , T. , Fujiyama , A. and Matsubara , K.Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA . Proc. Natl. Acad. Sci. USA , 84 , 444 – 448 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. ) Yaginuma , K. , Shirakata , Y. , Kobayashi , M. and Koike , K.Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA . Proc. Natl. Acad. Sci. USA , 84 , 2678 – 2682 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. ) Ochiya , T. , Tsurimoto , T. , Ueda , K. , Okubo , K. , Shiozawa , M. and Matsubara , K.An in vitro system for infection with hepatitis B virus that uses primary human fetal hepatocytes . Proc. Natl. Acad. Sci. USA , 86 , 1875 – 1879 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. ) Shaul , Y. , Rutter , W. J. and Laub , O.A human hepatitis B viral enhancer element . EMBO J. , 4 , 427 – 430 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12. ) Tognoni , A. , Cattaneo , R. , Serfling , E. and Schaffner , W.A novel expression selection approach allows precise mapping of the hepatitis B virus enhancer . Nucleic Acid Res. , 13 , 7454 – 7472 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. ) Yee , J‐K.A liver specific enhancer in the core promoter region of human hepatitis B virus . Science , 246 , 658 – 661 ( 1989. ). [DOI] [PubMed] [Google Scholar]
- 14. ) Burk , R. D. , DeLoia , J. A. , ElAwady , M. K. and Gearhart , J. D.Tissue preferential expression of the hepatitis B virus (HBV) surface antigen gene in two lines of HBV transgenic mice . J. Viral , 62 , 649 – 654 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. ) Moriyama , T. , Guilhot , S. , Klopchin , K. , Moss , B. , Pinkert , C. A. , Palmiter , R. D. , Brinster , R. L. , Kanagawa , O. and Chisari , F. V.Immunobiology and pathogenesis of hepatocellular injury in hepatitis B virus transgenic mice . Science , 248 , 361 – 364 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 16. ) Walker , M. E. , Szmuness , W. , Stevens , C. and Rubinstein , P.Genetics of anti‐HBs responsiveness. I. HLA‐DR7 and nonresponsiveness to hepatitis vaccination . Transfusion , 21 , 601 ( 1981. ). [Google Scholar]
- 17. ) Milich , D. R. , Leroux‐Roeis , G. G. and Chisari , F. V.Genetic regulation of the immune response to hepatitis B surface antigen (HBsAg). II. Qualitative characteristics of the humoral immune response to the a, d, and y determinants of HBsAg . J. ImmunoL , 130 , 1395 – 1400 ( 1983. ). [PubMed] [Google Scholar]