Construction and characterisation of a recombinant fowlpox virus that expresses the human papilloma virus L1 protein

J Transl Med. 2011 Nov 4:9:190. doi: 10.1186/1479-5876-9-190.

Abstract

Background: Human papilloma virus (HPV)-16 is the most prevalent high-risk mucosal genotype. Virus-like-particle (VLP)-based immunogens developed recently have proven to be successful as prophylactic HPV vaccines, but are still too expensive for developing countries. Although vaccinia viruses expressing the HPV-16 L1 protein (HPV-L1) have been studied, fowlpox-based recombinants represent efficient and safer vectors for immunocompromised hosts due to their ability to elicit a complete immune response and their natural host-range restriction to avian species.

Methods: A new fowlpox virus recombinant encoding HPV-L1 (FPL1) was engineered and evaluated for the correct expression of HPV-L1 in vitro, using RT-PCR, immunoprecipitation, Western blotting, electron microscopy, immunofluorescence, and real-time PCR assays.

Results: The FPL1 recombinant correctly expresses HPV-L1 in mammalian cells, which are non-permissive for the replication of this vector.

Conclusion: This FPL1 recombinant represents an appropriate immunogen for expression of HPV-L1 in human cells. The final aim is to develop a safe, immunogenic, and less expensive prophylactic vaccine against HPV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism*
  • Cell Line
  • Fluorescent Antibody Technique
  • Fowlpox virus / genetics*
  • Fowlpox virus / ultrastructure
  • Gene Expression Regulation
  • Genetic Techniques*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Human papillomavirus 16 / metabolism*
  • Humans
  • Immunoprecipitation
  • Mammals
  • Oncogene Proteins, Viral / genetics
  • Oncogene Proteins, Viral / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Recombination, Genetic / genetics*
  • Time Factors
  • Virion / metabolism
  • Virion / ultrastructure

Substances

  • Capsid Proteins
  • HPV L1 protein, Human papillomavirus
  • Oncogene Proteins, Viral
  • RNA, Messenger
  • Green Fluorescent Proteins