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. 1997 Oct;71(10):7381–7386. doi: 10.1128/jvi.71.10.7381-7386.1997

Synthesis of infectious human papillomavirus type 18 in differentiating epithelium transfected with viral DNA.

C Meyers 1, T J Mayer 1, M A Ozbun 1
PMCID: PMC192083  PMID: 9311816

Abstract

The lack of a permissive system for the propagation of viral stocks containing abundant human papillomavirus (HPV) particles has hindered the study of infectivity and the early stages of HPV replication. The organotypic (raft) culture system has permitted the study of a number of the differentiation-specific aspects of HPV, including amplification of viral DNA, expression of late genes, and viral morphogenesis. However, these investigations have been limited to a single virus type, namely, HPV type 31 (HPV31). We have artificially introduced linearized HPV18 genomic DNA into primary keratinocytes by electroporation, followed by clonal expansion and induction of epithelial stratification and differentiation in organotypic culture. We report the synthesis of infectious HPV18 virions. Virus particles approximately 50 nm in diameter were observed by electron microscopy. HPV18 virions purified by isopycnic gradient were capable of infecting keratinocytes in vitro, as shown by the expression of multiple HPV18-specific, spliced transcripts.

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Selected References

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  1. Atencio I. A., Villarreal L. P. Polyomavirus replicates in differentiating but not in proliferating tubules of adult mouse polycystic kidneys. Virology. 1994 May 15;201(1):26–35. doi: 10.1006/viro.1994.1262. [DOI] [PubMed] [Google Scholar]
  2. Choo K. B., Cheung W. F., Liew L. N., Lee H. H., Han S. H. Presence of catenated human papillomavirus type 16 episomes in a cervical carcinoma cell line. J Virol. 1989 Feb;63(2):782–789. doi: 10.1128/jvi.63.2.782-789.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crawford D. H., Ando I. EB virus induction is associated with B-cell maturation. Immunology. 1986 Nov;59(3):405–409. [PMC free article] [PubMed] [Google Scholar]
  4. Cullen B. R., Greene W. C. Regulatory pathways governing HIV-1 replication. Cell. 1989 Aug 11;58(3):423–426. doi: 10.1016/0092-8674(89)90420-0. [DOI] [PubMed] [Google Scholar]
  5. Davies A. H., Grand R. J., Evans F. J., Rickinson A. B. Induction of Epstein-Barr virus lytic cycle by tumor-promoting and non-tumor-promoting phorbol esters requires active protein kinase C. J Virol. 1991 Dec;65(12):6838–6844. doi: 10.1128/jvi.65.12.6838-6844.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Frattini M. G., Lim H. B., Laimins L. A. In vitro synthesis of oncogenic human papillomaviruses requires episomal genomes for differentiation-dependent late expression. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):3062–3067. doi: 10.1073/pnas.93.7.3062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hudson J. B., Bedell M. A., McCance D. J., Laiminis L. A. Immortalization and altered differentiation of human keratinocytes in vitro by the E6 and E7 open reading frames of human papillomavirus type 18. J Virol. 1990 Feb;64(2):519–526. doi: 10.1128/jvi.64.2.519-526.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Johnson P., Chung S., Benchimol S. Growth suppression of Friend virus-transformed erythroleukemia cells by p53 protein is accompanied by hemoglobin production and is sensitive to erythropoietin. Mol Cell Biol. 1993 Mar;13(3):1456–1463. doi: 10.1128/mcb.13.3.1456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaur P., McDougall J. K. HPV-18 immortalization of human keratinocytes. Virology. 1989 Nov;173(1):302–310. doi: 10.1016/0042-6822(89)90247-x. [DOI] [PubMed] [Google Scholar]
  11. Kiessling U., Kube D., Janda J., Bergmann S., Johannsmeyer K. D., Hauff P., Arnoldt W., Waehlte H. Properties of two epithelial cell lines derived from HPV-associated cervical and vulvar lesions. Eur J Cancer. 1993;29A(12):1746–1753. doi: 10.1016/0959-8049(93)90118-y. [DOI] [PubMed] [Google Scholar]
  12. Laurence J., Cooke H., Sikder S. K. Effect of tamoxifen on regulation of viral replication and human immunodeficiency virus (HIV) long terminal repeat-directed transcription in cells chronically infected with HIV-1. Blood. 1990 Feb 1;75(3):696–703. [PubMed] [Google Scholar]
  13. Laverty C. R., Russell P., Hills E., Booth N. The significance of noncondylomatous wart virus infection of the cervical transformation zone. A review with discussion of two illustrative cases. Acta Cytol. 1978 Jul-Aug;22(4):195–201. [PubMed] [Google Scholar]
  14. Lewis R. M., Morrill J. C., Jahrling P. B., Cosgriff T. M. Replication of hemorrhagic fever viruses in monocytic cells. Rev Infect Dis. 1989 May-Jun;11 (Suppl 4):S736–S742. doi: 10.1093/clinids/11.supplement_4.s736. [DOI] [PubMed] [Google Scholar]
  15. Li Q. X., Young L. S., Niedobitek G., Dawson C. W., Birkenbach M., Wang F., Rickinson A. B. Epstein-Barr virus infection and replication in a human epithelial cell system. Nature. 1992 Mar 26;356(6367):347–350. doi: 10.1038/356347a0. [DOI] [PubMed] [Google Scholar]
  16. McCance D. J., Kopan R., Fuchs E., Laimins L. A. Human papillomavirus type 16 alters human epithelial cell differentiation in vitro. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7169–7173. doi: 10.1073/pnas.85.19.7169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meyers C., Frattini M. G., Hudson J. B., Laimins L. A. Biosynthesis of human papillomavirus from a continuous cell line upon epithelial differentiation. Science. 1992 Aug 14;257(5072):971–973. doi: 10.1126/science.1323879. [DOI] [PubMed] [Google Scholar]
  18. Meyers C., Laimins L. A. In vitro systems for the study and propagation of human papillomaviruses. Curr Top Microbiol Immunol. 1994;186:199–215. doi: 10.1007/978-3-642-78487-3_11. [DOI] [PubMed] [Google Scholar]
  19. Morin C., Meisels A. Human papilloma virus infection of the uterine cervix. Acta Cytol. 1980 Jan-Feb;24(1):82–84. [PubMed] [Google Scholar]
  20. Ozbun M. A., Meyers C. Characterization of late gene transcripts expressed during vegetative replication of human papillomavirus type 31b. J Virol. 1997 Jul;71(7):5161–5172. doi: 10.1128/jvi.71.7.5161-5172.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ozbun M. A., Meyers C. Transforming growth factor beta1 induces differentiation in human papillomavirus-positive keratinocytes. J Virol. 1996 Aug;70(8):5437–5446. doi: 10.1128/jvi.70.8.5437-5446.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Parkin D. M., Pisani P., Ferlay J. Estimates of the worldwide incidence of eighteen major cancers in 1985. Int J Cancer. 1993 Jun 19;54(4):594–606. doi: 10.1002/ijc.2910540413. [DOI] [PubMed] [Google Scholar]
  23. Pilotti S., Rilke F., De Palo G., Della Torre G., Alasio L. Condylomata of the uterine cervix and koilocytosis of cervical intraepithelial neoplasia. J Clin Pathol. 1981 May;34(5):532–541. doi: 10.1136/jcp.34.5.532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Polyak S. J., Rawls W. E., Harnish D. G. Characterization of Pichinde virus infection of cells of the monocytic lineage. J Virol. 1991 Jul;65(7):3575–3582. doi: 10.1128/jvi.65.7.3575-3582.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schneider-Gädicke A., Schwarz E. Different human cervical carcinoma cell lines show similar transcription patterns of human papillomavirus type 18 early genes. EMBO J. 1986 Sep;5(9):2285–2292. doi: 10.1002/j.1460-2075.1986.tb04496.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schneider-Schaulies S., Schneider-Schaulies J., Bayer M., Löffler S., ter Meulen V. Spontaneous and differentiation-dependent regulation of measles virus gene expression in human glial cells. J Virol. 1993 Jun;67(6):3375–3383. doi: 10.1128/jvi.67.6.3375-3383.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shally M., Alloul N., Jackman A., Muller M., Gissmann L., Sherman L. The E6 variant proteins E6I-E6IV of human papillomavirus 16: expression in cell free systems and bacteria and study of their interaction with p53. Virus Res. 1996 Jun;42(1-2):81–96. doi: 10.1016/0168-1702(96)01301-9. [DOI] [PubMed] [Google Scholar]
  28. Small J. A., Bieberich C., Ghotbi Z., Hess J., Scangos G. A., Clements J. E. The visna virus long terminal repeat directs expression of a reporter gene in activated macrophages, lymphocytes, and the central nervous systems of transgenic mice. J Virol. 1989 May;63(5):1891–1896. doi: 10.1128/jvi.63.5.1891-1896.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Viac J., Schmitt D., Thivolet J. An immunoelectron microscopic localization of wart associated antigens present in human papilloma virus (HPV) infected cells. J Invest Dermatol. 1978 May;70(5):263–266. doi: 10.1111/1523-1747.ep12541415. [DOI] [PubMed] [Google Scholar]
  30. Visalli R. J., Courtney R. J., Meyers C. Infection and replication of herpes simplex virus type 1 in an organotypic epithelial culture system. Virology. 1997 Apr 14;230(2):236–243. doi: 10.1006/viro.1997.8484. [DOI] [PubMed] [Google Scholar]
  31. Weinshenker B. G., Wilton S., Rice G. P. Phorbol ester-induced differentiation permits productive human cytomegalovirus infection in a monocytic cell line. J Immunol. 1988 Mar 1;140(5):1625–1631. [PubMed] [Google Scholar]
  32. Wettstein F. O., Stevens J. G. Variable-sized free episomes of Shope papilloma virus DNA are present in all non-virus-producing neoplasms and integrated episomes are detected in some. Proc Natl Acad Sci U S A. 1982 Feb;79(3):790–794. doi: 10.1073/pnas.79.3.790. [DOI] [PMC free article] [PubMed] [Google Scholar]

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