Production and Prophylactic Efficacy Study of Human Papillomavirus-like Particle Expressing HPV16 L1 Capsid Protein

  • Published : 2002.12.01


To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported to induce significant humoral and cellular immunity using various animal model systems. In order to produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain reaction, was inserted into yeast expression vector, YEG$\alpha$-HIR525 under the control of GAL10 promoter. The transformation of YEG$\alpha$-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody. The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and simple assay to detect hemagglutihation inhibition activity.


  1. Seminars in Cancer Biol. v.9 Human papillomavirus vaccines. Breitburd,F.;P.Coursaget.
  2. Virology v.293 Yeast coexpression of human papillomavirus types 6 and 16 capsid protein. Buonamassa,D.T.;C.E.Greer;S.Capo;T.S.B.Yen;C.L.Galeotti;G.BENSI.
  3. Virology v.182 Expression fo human papillomavirus proteins in yeast Saccharomyees Cerevisiae. Carter,J.J.;N.Yaegashi;S.A.Jenison;D.A.Galloway.
  4. Cancer Lett. v.162 Improvement of gene transfer to cervical cancer cell lines using non-viral agents. Cho,C.W.;Y.S.Cho;B.T.Kang;J.S.Hwang;S.N.Park;D.Y.Yoon
  5. J. Infect. Dis. v.183 no.10 A Phase 1 study of a recombinant virus-like particle vaccine against human papillomavirus type 11 in healthy adult volunteers. Evans,T.G.;W.Bonnez;R.C.Rose;S.Koenig;L.Demeter;J.A.Suzich;D.O’Brien;M.Campbell;W.I.White;J.Balsley;R.C.Reichman
  6. J. Virol. Human papillomavirus virus-like particles are efficient oral immunogens when coadministered with Escherichia coli heat-labile Enterotoxin Mutant R192G or CpG DNA. Gerber,S.;C.Lane;D.M.Brown;E.Lord;M.Dilorenzo;J.D.Clements;E.Rybicki;A.L.Williamson;R.C.Rose.
  7. J.Virol. v.67 Self-assembiy of human papillomavirus type 1 capsids by expression of the L1 protein alone or by oexpression of the L1 and L2 capsid proteins. Hagensee,M.E.;N.Yaegashi;D.A.Galloway.
  8. J. Natl.Cancer Inst. v.93 no.4 Papillomavirus 16L1 virus-like paticle vaccine Harro,C.D.;Y.S.Pang;R.B.S.Rose;A.Hildesheim;Z.Wang;M.J.Reynolds;T.C.Mast;R.Robinson;B.R.Murphy;R.A.Karron;J.Dillner;T.Schiller;D.R.Lowy.
  9. Vaccine v.13 Vaccination with yeast-expressed cottontail rabbits papillomavirus(CRPV) virus-like particles protects rabbits from CRPV-induced papilloma formation. Jansen,K.U.;M.Rosolowsky;L.D.Schultz;H.Z.Markus;J.C.Cook;J.J.Donnelly;D.Martinez;R.W.Ellis;A.R.Shaw.
  10. Biomed. Pharmacother. v.53 Induction of cellular immunosuppression by the human papillomavirus type 16 E7 oncogenic protein. Le Buanec,H.;A.Lachger;R.D’Anna;J.F.Zagury;B.Bizzini;J.Bernard;D.Ittele;S.Hallez;C.Giannouli;A.Burny;D.Zaguty.
  11. J. Immunol.2001 v.166 Papillomavirus-like particles induce acute activation of dendritic cells. Lenz,P.;P.M.Day;Y.S.Pang;S.A.Frye;P.N.Jensen;D.R.Lowy;J.T.Schiller.
  12. Proc. Nalt. Acad.Sci.USA. v.91 Genital human papillomavirus infection. Lowy,D.R.;R.Kirnbauer;J.T.Schiller.
  13. J. Virol.1996 Assessment of the serological relatedness of genital human papillomaviruses by hemagglutination inhibition. Roden,R.A.S;N.L.Hubbert;R.Kirnbauer;N.D.Chridtensen;D.R.Lowy;J.T.Schiller.
  14. Virology v.270 Minor Capsid protein of human genital papillomaviruses contains subdominant, cross-neutalizing epitoper. Roden,R.B.S.;W.H.Yutzy;R.Fallon;S.Inglis;D.R.Lowy;J.T.Schiller.
  15. Hum. Gene Ther. v.11 Assembly of human papillomavirus type 16 pseudovirions in Saccharomyces cerevisiae. Rossi,J.L.;L.Gissmann;K.Jansen;M.Muller.
  16. J. Clinic. Virol. v.19 Deneloping HPV virus-like particle vaccines to prevent cervical cancer: a prohress report. Schiller,J.T.;A.Hidesheim.
  17. Seminars in Cancer Biol. v.7 Papillomavirus0like particles and HPV vaccine development. Schiller,J.T.;D.R.Lowy.
  18. J. Microbiol. v.40 Histological alteration and immune response inducde by pet toxin during colonization with enteroaggregative Escherichia coli(EAEC) in a mouse midel infection. Teresita, S.;J.Perez;M.C.Fresan;V.Flotes;L.Jimenez;U.Hernandez;I.Herrera;C.Eslava.
  19. Biochem. Biophys. Res.Commun. v.283 no.3 Alpha(6) Integrin id the main receptor of human papillomavirus type 16VLP. Yoon,C.S.;K.D.Kim,;S.N.Park;S.W.Cheong.
  20. Virology v.189 Definition of liner antigenic regions of the HPV16 L1 capsid protein using synthetic virion-like particles. Zhou,J.;X.Y.Sun;H.Davies;L.Crawford;D.Park;I.H.Fraxer.