Stably expressed FIPV peplomer protein induces cell fusion and elicits neutralizing antibodies in mice

Virology. 1989 Aug;171(2):493-502. doi: 10.1016/0042-6822(89)90619-3.

Abstract

We have established bovine papilloma virus (BPV)-transformed mouse C127 cell lines that synthesize the peplomer protein of the feline infectious peritonitis virus (FIPV) strain 79-1146. For this purpose, a new cassette expression vector pHSL, which carries the Drosophila HSp70 promotor and the polyadenylation signal of the Moloney murine leukemia virus long terminal repeat, was constructed. Cocultivation of the BPV-transformed cell lines with FIPV-permissive feline fcwf-D cells resulted in polykaryocyte formation. Since it depended on the presence of fcwf-D cells, binding of E2 to the cell receptor may be required for membrane fusion. E2 was synthesized as a core-glycosylated protein of 180K which was only slowly transported from the endoplasmic reticulum to the medial Golgi: of the E2-molecules labeled during a 1-hr pulse about half was still completely sensitive to endoglycosidase H after a 2-hr chase, while the remaining E2 had been chased into multiple, partially endoglycosidase H-resistant forms. Immunofluorescence studies also indicated that most E2 was retained intracellularly. Mice immunized with whole lysates of the transformed cells produced FIPV-neutralizing antibodies as shown by plaque reduction.

MeSH terms

  • Animals
  • Antibodies, Viral / immunology*
  • Biological Transport
  • Bovine papillomavirus 1 / genetics
  • Cell Fusion*
  • Cell Line
  • Cell Membrane / metabolism
  • Cloning, Molecular
  • Coronaviridae / genetics*
  • Coronaviridae / physiology
  • Genetic Vectors
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / immunology
  • Mice
  • Protein Processing, Post-Translational
  • Transfection
  • Viral Envelope Proteins / genetics*
  • Viral Envelope Proteins / immunology

Substances

  • Antibodies, Viral
  • Membrane Glycoproteins
  • Viral Envelope Proteins