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. 1993 Apr;67(4):1936–1944. doi: 10.1128/jvi.67.4.1936-1944.1993

Expression of human papillomavirus type 11 L1 protein in insect cells: in vivo and in vitro assembly of viruslike particles.

R C Rose 1, W Bonnez 1, R C Reichman 1, R L Garcea 1
PMCID: PMC240261  PMID: 8383219

Abstract

The L1 coat protein of human papillomavirus type 11 (HPV-11) was expressed in Sf-9 insect cells with the recombinant baculovirus vector Ac11L1. Viruslike particles (VLPs) were identified by electron microscopy in the nucleus and cytoplasm of Sf-9 cells infected with Ac11L1. The L1 protein was purified from Ac11L1-infected insect cells. The purified protein spontaneously assembled in vitro into various aggregates, including particles appearing similar to empty virions. Reaction of VLP-containing insect cell extracts with antisera directed against either denatured or nondenatured capsid epitopes in Western blot (immunoblot) and immuno-dot blot assays suggested that conformational epitopes present in native HPV-11 infectious virions were also present on the baculovirus-produced HPV-11 VLPs. Immuno-dot blot assays using human sera obtained from individuals with biopsy-proven condyloma acuminatum correlated closely with results previously obtained in HPV-11 whole virus particle-based enzyme-linked immunosorbent assays. These morphologic and immunologic similarities to native HPV-11 virions suggest that recombinant VLPs produced in the baculovirus system may be useful in seroepidemiology and pathogenesis studies of genital HPV infection and that they may also be potential candidates for vaccine development.

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Selected References

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  1. Baker T. S., Newcomb W. W., Olson N. H., Cowsert L. M., Olson C., Brown J. C. Structures of bovine and human papillomaviruses. Analysis by cryoelectron microscopy and three-dimensional image reconstruction. Biophys J. 1991 Dec;60(6):1445–1456. doi: 10.1016/S0006-3495(91)82181-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonnez W., Da Rin C., Rose R. C., Reichman R. C. Use of human papillomavirus type 11 virions in an ELISA to detect specific antibodies in humans with condylomata acuminata. J Gen Virol. 1991 Jun;72(Pt 6):1343–1347. doi: 10.1099/0022-1317-72-6-1343. [DOI] [PubMed] [Google Scholar]
  3. Bonnez W., Rose R. C., Reichman R. C. Antibody-mediated neutralization of human papillomavirus type 11 (HPV-11) infection in the nude mouse: detection of HPV-11 mRNAs. J Infect Dis. 1992 Feb;165(2):376–380. doi: 10.1093/infdis/165.2.376. [DOI] [PubMed] [Google Scholar]
  4. Christensen N. D., Kreider J. W. Antibody-mediated neutralization in vivo of infectious papillomaviruses. J Virol. 1990 Jul;64(7):3151–3156. doi: 10.1128/jvi.64.7.3151-3156.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Christensen N. D., Kreider J. W., Cladel N. M., Patrick S. D., Welsh P. A. Monoclonal antibody-mediated neutralization of infectious human papillomavirus type 11. J Virol. 1990 Nov;64(11):5678–5681. doi: 10.1128/jvi.64.11.5678-5681.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christensen N. D., Kreider J. W., Shah K. V., Rando R. F. Detection of human serum antibodies that neutralize infectious human papillomavirus type 11 virions. J Gen Virol. 1992 May;73(Pt 5):1261–1267. doi: 10.1099/0022-1317-73-5-1261. [DOI] [PubMed] [Google Scholar]
  7. Crum C. P., Levine R. U. Human papillomavirus infection and cervical neoplasia: new perspectives. Int J Gynecol Pathol. 1984;3(4):376–388. [PubMed] [Google Scholar]
  8. Dun A. E., Ogilvie M. M. Intranuclear virus particles in human genital wart tissue: observations on the ultrastructure of the epidermal layer. J Ultrastruct Res. 1968 Feb;22(3):282–295. doi: 10.1016/s0022-5320(68)90021-x. [DOI] [PubMed] [Google Scholar]
  9. Gheysen D., Jacobs E., de Foresta F., Thiriart C., Francotte M., Thines D., De Wilde M. Assembly and release of HIV-1 precursor Pr55gag virus-like particles from recombinant baculovirus-infected insect cells. Cell. 1989 Oct 6;59(1):103–112. doi: 10.1016/0092-8674(89)90873-8. [DOI] [PubMed] [Google Scholar]
  10. Jenison S. A., Firzlaff J. M., Langenberg A., Galloway D. A. Identification of immunoreactive antigens of human papillomavirus type 6b by using Escherichia coli-expressed fusion proteins. J Virol. 1988 Jun;62(6):2115–2123. doi: 10.1128/jvi.62.6.2115-2123.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KLUG A., FINCH J. T. STRUCTURE OF VIRUSES OF THE PAPILLOMA-POLYOMA TYPE. I. HUMAN WART VIRUS. J Mol Biol. 1965 Feb;11:403–423. doi: 10.1016/s0022-2836(65)80066-3. [DOI] [PubMed] [Google Scholar]
  12. Kennedy I. M., Haddow J. K., Clements J. B. A negative regulatory element in the human papillomavirus type 16 genome acts at the level of late mRNA stability. J Virol. 1991 Apr;65(4):2093–2097. doi: 10.1128/jvi.65.4.2093-2097.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kennedy I. M., Haddow J. K., Clements J. B. Analysis of human papillomavirus type 16 late mRNA 3' processing signals in vitro and in vivo. J Virol. 1990 Apr;64(4):1825–1829. doi: 10.1128/jvi.64.4.1825-1829.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kienzler J. L., Lemoine M. T., Orth G., Jibard N., Blanc D., Laurent R., Agache P. Humoral and cell-mediated immunity to human papillomavirus type 1 (HPV-1) in human warts. Br J Dermatol. 1983 Jun;108(6):665–672. doi: 10.1111/j.1365-2133.1983.tb01078.x. [DOI] [PubMed] [Google Scholar]
  15. Kirnbauer R., Booy F., Cheng N., Lowy D. R., Schiller J. T. Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12180–12184. doi: 10.1073/pnas.89.24.12180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koutsky L. A., Galloway D. A., Holmes K. K. Epidemiology of genital human papillomavirus infection. Epidemiol Rev. 1988;10:122–163. doi: 10.1093/oxfordjournals.epirev.a036020. [DOI] [PubMed] [Google Scholar]
  17. Kreider J. W., Howett M. K., Leure-Dupree A. E., Zaino R. J., Weber J. A. Laboratory production in vivo of infectious human papillomavirus type 11. J Virol. 1987 Feb;61(2):590–593. doi: 10.1128/jvi.61.2.590-593.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kreider J. W., Howett M. K., Wolfe S. A., Bartlett G. L., Zaino R. J., Sedlacek T., Mortel R. Morphological transformation in vivo of human uterine cervix with papillomavirus from condylomata acuminata. Nature. 1985 Oct 17;317(6038):639–641. doi: 10.1038/317639a0. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Li C. C., Shah K. V., Seth A., Gilden R. V. Identification of the human papillomavirus type 6b L1 open reading frame protein in condylomas and corresponding antibodies in human sera. J Virol. 1987 Sep;61(9):2684–2690. doi: 10.1128/jvi.61.9.2684-2690.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. López de Turiso J. A., Cortés E., Martínez C., Ruiz de Ybáez R., Simarro I., Vela C., Casal I. Recombinant vaccine for canine parvovirus in dogs. J Virol. 1992 May;66(5):2748–2753. doi: 10.1128/jvi.66.5.2748-2753.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Montross L., Watkins S., Moreland R. B., Mamon H., Caspar D. L., Garcea R. L. Nuclear assembly of polyomavirus capsids in insect cells expressing the major capsid protein VP1. J Virol. 1991 Sep;65(9):4991–4998. doi: 10.1128/jvi.65.9.4991-4998.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rose R. C., Bonnez W., Strike D. G., Reichman R. C. Expression of the full-length products of the human papillomavirus type 6b (HPV-6b) and HPV-11 L2 open reading frames by recombinant baculovirus, and antigenic comparisons with HPV-11 whole virus particles. J Gen Virol. 1990 Nov;71(Pt 11):2725–2729. doi: 10.1099/0022-1317-71-11-2725. [DOI] [PubMed] [Google Scholar]
  24. Roy P., French T., Erasmus B. J. Protective efficacy of virus-like particles for bluetongue disease. Vaccine. 1992;10(1):28–32. doi: 10.1016/0264-410x(92)90415-g. [DOI] [PubMed] [Google Scholar]
  25. Sabara M., Parker M., Aha P., Cosco C., Gibbons E., Parsons S., Babiuk L. A. Assembly of double-shelled rotaviruslike particles by simultaneous expression of recombinant VP6 and VP7 proteins. J Virol. 1991 Dec;65(12):6994–6997. doi: 10.1128/jvi.65.12.6994-6997.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Saliki J. T., Mizak B., Flore H. P., Gettig R. R., Burand J. P., Carmichael L. E., Wood H. A., Parrish C. R. Canine parvovirus empty capsids produced by expression in a baculovirus vector: use in analysis of viral properties and immunization of dogs. J Gen Virol. 1992 Feb;73(Pt 2):369–374. doi: 10.1099/0022-1317-73-2-369. [DOI] [PubMed] [Google Scholar]
  27. Salunke D. M., Caspar D. L., Garcea R. L. Polymorphism in the assembly of polyomavirus capsid protein VP1. Biophys J. 1989 Nov;56(5):887–900. doi: 10.1016/S0006-3495(89)82735-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Steele J. C., Gallimore P. H. Humoral assays of human sera to disrupted and nondisrupted epitopes of human papillomavirus type 1. Virology. 1990 Feb;174(2):388–398. doi: 10.1016/0042-6822(90)90092-6. [DOI] [PubMed] [Google Scholar]
  30. Strike D. G., Bonnez W., Rose R. C., Reichman R. C. Expression in Escherichia coli of seven DNA fragments comprising the complete L1 and L2 open reading frames of human papillomavirus type 6b and localization of the 'common antigen' region. J Gen Virol. 1989 Mar;70(Pt 3):543–555. doi: 10.1099/0022-1317-70-3-543. [DOI] [PubMed] [Google Scholar]
  31. Urakawa T., Ferguson M., Minor P. D., Cooper J., Sullivan M., Almond J. W., Bishop D. H. Synthesis of immunogenic, but non-infectious, poliovirus particles in insect cells by a baculovirus expression vector. J Gen Virol. 1989 Jun;70(Pt 6):1453–1463. doi: 10.1099/0022-1317-70-6-1453. [DOI] [PubMed] [Google Scholar]
  32. Xi S. Z., Banks L. M. Baculovirus expression of the human papillomavirus type 16 capsid proteins: detection of L1-L2 protein complexes. J Gen Virol. 1991 Dec;72(Pt 12):2981–2988. doi: 10.1099/0022-1317-72-12-2981. [DOI] [PubMed] [Google Scholar]
  33. Zhou J., Sun X. Y., Stenzel D. J., Frazer I. H. Expression of vaccinia recombinant HPV 16 L1 and L2 ORF proteins in epithelial cells is sufficient for assembly of HPV virion-like particles. Virology. 1991 Nov;185(1):251–257. doi: 10.1016/0042-6822(91)90772-4. [DOI] [PubMed] [Google Scholar]

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