Human-like antibodies neutralizing Western equine encephalitis virus

MAbs. May-Jun 2014;6(3):718-27. doi: 10.4161/mabs.28170. Epub 2014 Feb 11.

Abstract

This study describes the development of the first neutralizing antibodies against Western equine encephalitis virus (WEEV), a member of the genus Alphavirus. WEEV is transmitted by mosquitoes and can spread to the human central nervous system, causing symptoms ranging from mild febrile reactions to life-threatening encephalitis. WEEV has been classified as a biological warfare agent by the US Centers for Disease Control and Prevention. No anti-WEEV drugs are currently commercially available. Neutralizing antibodies are useful for the pre- and post-exposure treatment of WEEV infections. In this study, two immune antibody gene libraries were constructed from two macaques immunized with inactivated WEEV. Four antibodies were selected from these libraries and recloned as scFv-Fc, with a human Fc part. These antibodies bound WEEV specifically in ELISA with little or no cross-reaction with other alphaviruses. They were further analyzed by immunohistochemistry. All binders were suitable for the intracellular detection of WEEV particles. Neutralizing activity was determined in vitro. Three of the four antibodies were found to be neutralizing; about 1 ng/mL of the best antibody (ToR69-3A2) neutralized 50% of 5x10(4) TCID 50/mL. Due to its human-like nature with a germinality index of 89% (VH) and 91% (VL), the ToR69-3A2 antibody is a promising candidate for future passive vaccine development.

Keywords: Alphavirus; NHP antibodies; Western equine encephalitis virus (WEEV); immune antibody library; passive vaccine; phage display; scFv; scFv-Fc.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Neutralizing / genetics
  • Antibodies, Neutralizing / immunology*
  • Antibodies, Viral / genetics
  • Antibodies, Viral / immunology*
  • Cloning, Molecular
  • Cross Reactions
  • Encephalitis Virus, Western Equine / immunology*
  • Encephalitis Virus, Western Equine / pathogenicity
  • Encephalomyelitis, Equine / immunology
  • Encephalomyelitis, Equine / prevention & control
  • Humans
  • Immunization
  • Immunization, Passive
  • Macaca fascicularis
  • Male
  • Peptide Library
  • Post-Exposure Prophylaxis
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology

Substances

  • Antibodies, Monoclonal
  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Peptide Library
  • Single-Chain Antibodies