Canine parvovirus empty capsids produced by expression in a baculovirus vector: use in analysis of viral properties and immunization of dogs.
The VP-2 genes of canine parvovirus (CPV) and a recombinant consisting of CPV and feline panleukopenia virus (FPV) sequences were cloned into baculovirus expression vectors, fused to the baculovirus polyhedrin promoter. Recombinant baculoviruses were prepared and the properties of the parvovirus proteins expressed in insect cells examined. The proteins produced were the same size as the authentic CPV VP-2 protein, and were produced late after infection; the quantity of proteins recovered from the insect cell cultures was similar to those produced in CPV infections. Parvovirus particles formed had the haemagglutination (HA), sedimentation and buoyant density properties of authentic CPV capsids. Both the CPV capsids and the CPV-FPV recombinant capsids from the baculovirus system expressed the same epitopes as those seen in the viable parvoviruses when tested with a panel of anti-parvovirus monoclonal antibodies. Lysates of recombinant baculovirus-infected cells were inoculated into dogs, giving rise to serum neutralizing and HA-inhibiting antibodies, and the immunized dogs were protected from clinical disease upon challenge with a virulent isolate of the most recent antigenic type of CPV.
James A. Baker Institute, New York State College of Veterinary Medicine, Cornell University, Ithaca 14853.
J T
JT
Saliki
B
B
Mizak
H P
HP
Flore
R R
RR
Gettig
J P
JP
Burand
L E
LE
Carmichael
H A
HA
Wood
C R
CR
Parrish
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002213:Capsid / Q000235:genetics* / Q000276:immunology; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral / Q000737:chemistry; D004283:Dog Diseases / Q000517:prevention & control; D004285:Dogs; D000939:Epitopes / Q000276:immunology; D015870:Gene Expression; D005822:Genetic Vectors; D007114:Immunization / Q000662:veterinary*; D008969:Molecular Sequence Data; D010321:Parvoviridae / Q000235:genetics* / Q000276:immunology; D010322:Parvoviridae Infections / Q000517:prevention & control / Q000662:veterinary; D010957:Plasmids; D014614:Vaccines, Synthetic; D014765:Viral Vaccines
D000914:Antibodies, Viral; D004279:DNA, Viral; D000939:Epitopes; D014614:Vaccines, Synthetic; D014765:Viral Vaccines