Baculovirus expression of the avian paramyxovirus 2 HN gene for diagnostic applications

J Virol Methods. 2014 Mar:198:12-7. doi: 10.1016/j.jviromet.2013.12.010. Epub 2013 Dec 24.

Abstract

Avian paramyxovirus 2 (APMV-2) infections are associated with respiratory diseases in poultry worldwide. The hemagglutination inhibition (HI) test is a useful tool for surveillance and monitoring of this virus. In this study, full-length hemagglutinin (HN) gene of APMV-2 was chemically synthesized based on its published sequence, cloned and expressed in Spodoptera frugiperda insect cells using recombinant baculoviruses. The biological, antigenic and immunogenic properties of the expressed protein were evaluated to assess its ability to produce diagnostic reagents for HI testing. Recombinant APMV-2 HN protein showed two distinct bands with molecular masses of 64 and 75kDa, which showed hemagglutination (HA) and neuraminidase activities, respectively. The recombinant HN (rHN) protein extracted from infected cells produced high HA titers (2(13) per 25μL). HA activity of the protein was inhibited by APMV-2 antiserum, although there were weak cross reactions with other APMV serotype antisera. The rHN protein induced high titers of APMV-2-specific antibodies in immunized chickens based on the HI test. These results indicated that recombinant APMV-2 HN protein is a useful alternative to the APMV-2 antigen in HI assays.

Keywords: Avian paramyxovirus type 2; Baculovirus expression; Hemagglutinin–neuraminidase protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Viral / immunology
  • Antigens, Viral / genetics
  • Antigens, Viral / immunology
  • Avulavirus / genetics*
  • Avulavirus / immunology
  • Avulavirus Infections / immunology
  • Avulavirus Infections / prevention & control
  • Avulavirus Infections / virology
  • Baculoviridae / genetics*
  • Base Sequence
  • Cell Line
  • Chickens / immunology
  • Chickens / virology
  • Cloning, Molecular / methods
  • Cross Reactions / immunology
  • HN Protein / genetics*
  • HN Protein / immunology
  • Hemagglutination Inhibition Tests / methods
  • Immune Sera / immunology
  • Molecular Sequence Data
  • Poultry Diseases / immunology
  • Poultry Diseases / prevention & control
  • Poultry Diseases / virology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sf9 Cells
  • Spodoptera / genetics

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • HN Protein
  • Immune Sera
  • Recombinant Proteins