1887

Journal of General Virology header logo

Volume 72, Issue 4

Baculovirus-directed high level expression of the hepatitis delta antigen in cells Free

Abstract

The hepatitis delta antigen (HDAg) is a multifunctional protein. It forms the core-like structure of the hepatitis delta virus (HDV) but also enhances replication of HDV in the nucleus of the hepatocyte. A cDNA fragment encoding HDAg was inserted adjacent to the polyhedrin promoter of nuclear polyhedrosis virus present in the baculovirus transfer vector pVL941. After transfection of (Sf9) cells a recombinant baculovirus Acδ1 was isolated and purified using filter hybridization techniques. Sf9 cells infected with Acδ1 express the HDAg as a non-fused, non-glycosylated protein with an abundance of up to 25% of the total cellular protein mass. Immunoblot analysis using a human polyclonal anti-HD conjugate identified a 22K and a 24K protein in the nucleus of Acδ1-infected Sf9 cells. Electron microscopic studies using immunogold labelling showed that the recombinant HDAg (recHDAg) was associated with the hetero-chromatin of the Sf9 cells. The recHDAg produced by Sf9 cells elicited anti-HD antibodies in chimpanzees when injected intramuscularly.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1991
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-4-833
1991-04-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/4/JV0720040833.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-4-833&mimeType=html&fmt=ahah

References

  1. Bergmann K. L., Gerin J. L. 1986; Antigens of hepatitis delta virus in the liver and serum of humans and animals. Journal of Infectious Diseases 154:702–706
     [Google Scholar]
  2. Bergmann K. L., Cote J. L., Moriarity A., Gerin J. L. 1989; Antigenic structure and humoral immune response. Journal of Immunology 143:3714–3721
     [Google Scholar]
  3. Chang M.-F., Baker S. C., Soe L. H., Kamahora T., Keck J. G., Makino S., Govindarajan S., Lai M. C. 1988; Human hepatitis delta antigen is a nuclear phosphoprotein with RNA-binding activity. Journal of Virology 62:2403–2410
     [Google Scholar]
  4. Chen P.-J., Kalpana G., Goldberg J., Mason W., Werner B., Gerin J. L., Taylor J. M. 1986; Structure and replication of the genome of the hepatitis δ virus. Proceedings of the National Academy of Sciences, U,. S,. A 83:8774–8778
     [Google Scholar]
  5. Fung M.-C., Chiu Y. M., Weber T., Chang T.-W., Chang N. T. 1988; Detection and purification of a recombinant human B lymphotrophic vims (HHV-6) in the baculovirus expression system by limiting dilution and DNA dot-blot hybridization. Journal of Virological Methods 19:33–42
     [Google Scholar]
  6. Glenn J. S., Taylor J. M., White J. M. 1990; In vitro synthesized hepatitis delta virus RNA initiates genome replication in cultured cells. Journal of Virology 64:3104–3107
     [Google Scholar]
  7. Hsieh S.-Y., Chao M., Coates L., Taylor J. 1990; Hepatitis delta vims genome replication: a polyadenylated mRNA for delta antigen. Journal of Virology 64:3192–3198
     [Google Scholar]
  8. Kos A., Dijkema R., Arnberg A. C., Van Der Meide P. H., Schellekens H. 1986; The HDV possesses a circular RNA. Nature, London 323:558–560
     [Google Scholar]
  9. Kuo M. Y.-P., Goldberg J., Coates F., Mason W., Gerin J. F., Taylor J. M. 1988; Molecular cloning of hepatitis delta vims RNA from an infected woodchuck liver: sequence, structure and applications. Journal of Virology 62:1855–1861
     [Google Scholar]
  10. Kuo M. Y.-P., Chao M., Taylor J. 1989; Initiation of replication of the human hepatitis delta vims genome from cloned DNA: role of delta antigen. Journal of Virology 63:1945–1950
     [Google Scholar]
  11. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
     [Google Scholar]
  12. Luckow V. A., Summers M. D. 1989; High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology 170:31–39
     [Google Scholar]
  13. Luo G., Chao M., Hsieh S.-Y., Sureau C., Nishikura K., Taylor J. 1990; A specific base transition occurs on replicating hepatitis delta vims RNA. Journal of Virology 64:1021–1027
     [Google Scholar]
  14. Macnaughton T. B., Gowans E. J., Reinboth B., Jilbert A. R., Burrell C. J. 1990; Stable expression of hepatitis delta virus antigen in a eukaryotic cell line. Journal of General Virology 71:1339–1345
     [Google Scholar]
  15. Makino S. M., Chang M., Shieh C., Kamahora T., Vannier D., Govindarajan S., Lai M. C. 1987; Molecular cloning and sequencing of a human hepatitis delta vims RNA. Nature, London 329:343–346
     [Google Scholar]
  16. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  17. Negro F., Gerin J. L. 1989b; Basis of hepatitis delta vims disease?. Nature, London 341:111
     [Google Scholar]
  18. Negro F., Shapiro M., Satterfield W. C., Gerin J. L., Purcell R. H. 1989a; Reappearance of hepatitis D virus (HDV) replication in chronic hepatitis B carrier chimpanzees rechallenged with HDV. Journal of Infectious Diseases 160:567–571
     [Google Scholar]
  19. Pen J., Welling G. W., Welling-Wester S. 1989; An efficient procedure for the isolation of recombinant baculovims. Nucleic Acids Research 17:451
     [Google Scholar]
  20. Ponzetto A., Forzani B., Smedile A., Hele C., Avanzini L., Novara R., Canese M. G. 1987a; Acute and chronic delta infection in the woodchuck. In The Hepatitis Delta Virus And Its Infection pp 37–46 Edited by Rizetto M., Gerin J. L., Purcell R. H. New York: Alan R. Liss;
     [Google Scholar]
  21. Ponzetto A., Rapicetta M., Forzani B., Smedile A., Hele C., Morace G., Di Rienzo A. M., Palladino P., Avanzini L., Gerin J. L. 1987b; Hepatitis delta vims infection in Pekin ducks chronically infected by the duck hepatitis B virus. In The Hepatitis Delta Virus And Its Infection pp 47–49 Edited by Rizetto M., Gerin J. L., Purcell R. H. New York: Alan R. Liss;
     [Google Scholar]
  22. Rizetto M., Canese M. G., Arico J., Crivelli O., Bonino F., Trepo C. G., Verme G. 1977; Immunofluorescence detection of a new antigen-antibody system (δ/anti-δ) associated to the hepatitis B virus in the liver and in the serum of HBsAg carriers. Gut 18:997–1003
     [Google Scholar]
  23. Rizetto M., Gerin J. L., Purcell R. H. 1981; Delta antigen: evidence for a variant of hepatitis B virus or non-A non-B hepatitis agent. Perspectives in Virology 11:195–217
     [Google Scholar]
  24. Roggendorf M., Pahlke C., Böhm B., Rasshofer R. 1987; Characterization of proteins associated with hepatitis delta virus. Journal of General Virology 68:2953–2959
     [Google Scholar]
  25. Saldanha J., Homer E., Goldin R., Thomas H. C., Monjardino J. 1990a; Cloning and expression of an immunodominant region of the hepatitis delta antigen. Journal of General Virology 71:471–475
     [Google Scholar]
  26. Saldanha J. A., Thomas H. C., Monjardino J. P. 1990b; Cloning and sequencing of RNA of hepatitis delta vims isolated from human semm. Journal of General Virology 71:1603–1606
     [Google Scholar]
  27. Sharmeen L., Kuo M. Y.-P., Dinter-Gottlieb G., Taylor J. 1988; Antigenomic RNA of human hepatitis delta vims can undergo self-cleavage. Journal of Virology 62:2674–2679
     [Google Scholar]
  28. Slot J. W., Geuze H. J., Freeman B. A., Crapo J. D. 1986; Intracellular localization of the copper-zinc and manganese superoxide dismutases in rat liver parenchymal cells. Laboratory Investigations 55:363–371
     [Google Scholar]
  29. Smith G. E., Summers M. D., Fraser M. J. 1983; Production of human beta interferon in insect cells infected with a baculovims expression vector. Molecular and Cellular Biology 3:2156–2165
     [Google Scholar]
  30. Stiles B., Wood H. A., Hughes P. R. 1983; Effect of tunicamycin on the infectivity of Autographa californica nuclear polyhedrosis virus. Journal of Invertebrate Pathology 41:405–408
     [Google Scholar]
  31. Summers M. D., Smith G. E. 1987; A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experimental Station Bulletin 1555:1–55
     [Google Scholar]
  32. Sureau C., Taylor J. M., Chao M., Eichberg J. W., Landford R. E. 1989; Cloned hepatitis delta virus cDNA is infectious in the chimpanzee. Journal of Virology 63:4292–4297
     [Google Scholar]
  33. Takatsuki A., Kohno K., Tamura G. 1975; Inhibition of biosynthesis of polyisoprenol sugars in chick embryo microsomes by tunicamycin. Agricultural Biological Chemistry 39:2089–2091
     [Google Scholar]
  34. Taylor J. M. 1990; Hepatitis delta virus cis and trans functions required for replication. Cell 61:371–373
     [Google Scholar]
  35. Tokuyasu K. T. 1980; Immunohistochemistry on ultrathin frozen sections. Histochemical Journal 12:381–403
     [Google Scholar]
  36. Wang J.-G., Jansen R. W., Brown A. E., Lemon S. M. 1990; Immunogenic domains of hepatitis delta virus antigen: peptide mapping of epitopes recognized by human and woodchuck antibodies. Journal of Virology 64:1108–1116
     [Google Scholar]
  37. Wang K.-S., Choo Q.-L., Weiner A. J., Ou H.-J., Najarian R. C., Thayer R. M., Mullenbach G. T., Denniston K. J., Gerin J. L., Houghton M. 1986; Structure, sequence and expression of the HDV genome. Nature, London 323:508–513
     [Google Scholar]
  38. Wang K.-S., Choo Q.-L., Weiner A. J., Ou H.-J., Najarian R. C., Thayer R. M., Mullenbach G. T., Denniston K. J., Gerin J. L., Houghton M. 1987; Structure, sequence and expression of the HDV genome. Nature, London 328:456
     [Google Scholar]
  39. Weiner A. J., Choo Q.-L., Wang K.-S., Govindarajan S., Redeker A. G., Gerin J. L., Houghton M. 1988; A single antigenomic open reading frame of the hepatitis delta virus encodes the epitope(s) of both hepatitis delta antigen polypeptides p 24 s and p 27δ . Journal of Virology 62:594–599
     [Google Scholar]
  40. Xia Y.-P., Chang M.-F., Wei D., Govindarajan S., Lai M. M. C. 1990; Heterogeneity of hepatitis delta antigen. Virology 178:331–336
     [Google Scholar]
  41. Young B., Hicke B. 1990; Delta virus as a cleaver. Nature, London 343:28
     [Google Scholar]
  42. Zyzik E., Ponzetto A., Forzani B., Hele C., Heermann K.-H., Gerlich W. H. 1987; Proteins of hepatitis delta virus in serum and liver. In Hepadnaviruses pp 565–577 Edited by Robinson W., Koike K., Will H. New York: Alan R. Liss;
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-4-833
Loading
Baculovirus-directed high level expression of the hepatitis delta antigen in Spodoptera frugiperda cells
J. Gen. Virol. 72, 833 (1991); https://doi.org/10.1099/0022-1317-72-4-833
/content/journal/jgv/10.1099/0022-1317-72-4-833
/content/journal/jgv/10.1099/0022-1317-72-4-833
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error