A deletion and point mutation study of the human papillomavirus type 16 major capsid gene

Arvind Varsani, Anna Lise Williamson, Mohamed A. Jaffer, Edward P. Rybicki

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Recombinant human papillomavirus (HPV) virus-like particles (VLPs) made from the major capsid protein L1 are promising vaccine candidates for use as vaccines against genital and other HPV infections, and particularly against HPV-16. However, HPV-16 genotype variants have different binding affinities for neutralising mouse Mabs raised against HPV-16 L1 VLPs. This paper analyses, using a panel of well-characterised Mabs, the effects on the antigenicity of various C- and N-terminal deletants of HPV-16 L1 made in insect cells via recombinant baculovirus, of an A → T mutation at residue 266 (A266T), and of a C → G mutation at conserved position 428 (C428G). The effects of these changes on assembly of the variant L1s were studied by electron microscopy. Binding of Mab H16:E70 to A266T was reduced by almost half in comparison to wild type L1. Retention of the C-terminal region 428-483 was critical for the binding of conformation-specific Mabs (H16:V5, H16:E70, H16:U4 and H16:9A) whereas deletion of the nuclear localisation signal (NLS) or the C428G mutation or an N-terminal deletion (residues 2-9) did not affect the antigenicity. The N-terminal deletion resulted in a mixed population of 30 and 55 nm VLPs, which differs from the same construct expressed in Escherichia coli, whereas pentamer aggregates resulted from deletion of the 428-465 region or the C428G mutation. The results have implications both for considering use of single-genotype HPV vaccines, and for design of novel second-generation vaccines.

Original languageEnglish (US)
Pages (from-to)154-163
Number of pages10
JournalVirus research
Volume122
Issue number1-2
DOIs
StatePublished - Dec 1 2006

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Keywords

  • Antigenicity
  • Deletion
  • HPV-16
  • L1
  • Papillomavirus
  • VLP

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

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