Heparin binding activity of orf virus F1L protein.
The orf virus is the type species of the Parapoxvirus genus and is the causative agent of contagious echtyma, a debilitating skin disease of sheep and goats, which can also affect man. The virus exhibits a restricted host range, even if it has been shown to bind to a wide range of tissues of non-permissive species. This ability is an argument for its potential use as an expression vector. Since most mammalian cell types express heparan sulfate (HS) surface receptors, we assumed that HS could serve as receptors to mediate orf virus binding. In this study, we showed that orf virus is inhibited by the addition of soluble heparin in cell cultures. Affinity chomatography using heparin agarose demonstrated that orf virus F1L is the major heparin binding protein. Furthermore, the recombinant F1L protein was visualised on the cell surface by confocal microscopy, and rabbits immunised with recombinant F1L protein produced virus neutralising antibodies. These results confirm that the F1L immunodominant protein is also involved in virus binding to cells as for the vaccinia homologue H3L protein. Heparin also inhibited the binding of the F1L protein to cells showing that this protein has a role in the early stages of infection.
Scagliarini
A
A
Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale, Alma Mater Studiorum, Bologna, Via Tolara di Sopra, 50, 40064 Ozzano Emilia Bologna, Italy. ascagliarini@vet.unibo.it
Gallina
L
L
Dal Pozzo
F
F
Battilani
M
M
Ciulli
S
S
Prosperi
S
S
D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology*; D018613:Microscopy, Confocal; D009500:Neutralization Tests; D009923:Orf virus / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D011485:Protein Binding; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000998:Antiviral Agents; D014764:Viral Proteins; D006493:Heparin