Induction of antibodies to plant viral proteins by DNA-based immunization.
DNA-based immunization is a promising new technique for generating antibodies in laboratory animals for diagnostic purposes in biological science. The main advantages are the elimination of time and labor and the technically demanding steps of antigen purification. The DNA sequence of the protein of interest, cloned in a suitable in vivo expression vector that is administered intramuscularly or intradermally, is sufficient to induce an immune response in animals. We report the induction of antibodies to tobacco mosaic virus (TMV) coat protein (CP) as a highly immunogenic structural protein and potato virus Y (PVY) P1 protein (P1) as a nonstructural protein. The appropriate nucleotide sequences were introduced in a mammalian expression vector (pSG5) and injected intramuscularly into New Zealand White rabbits (Oryctolagus cuniculus). By 10 days post-injection (dpi) a specific immune response was detected against TMV-CP, while it took about 5 weeks for a response to PVY P1. In both cases the antibody titers were significantly above the corresponding pre-immune serum, however, they were considerably below the titer of the matching conventionally produced antiserum. To our knowledge, this is the first report of DNA-based immunization in order to generate antibodies to plant viral proteins, but further improvements are necessary to increase antibody titers before this promising new technique can be introduced broadly in plant science for diagnostic purposes.
Hinrichs
J
J
University of Kentucky, College of Agriculture, Lexington 40546-0091, USA.
Berger
S
S
Shaw
J G
JG
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000918:Antibody Specificity; D002213:Capsid / Q000235:genetics / Q000276:immunology; D004279:DNA, Viral / Q000008:administration & dosage*; D007273:Injections, Intramuscular; D009029:Mosaic Viruses / Q000235:genetics / Q000276:immunology*; D017800:Potyvirus / Q000235:genetics / Q000276:immunology; D011817:Rabbits; D011993:Recombinant Fusion Proteins; D014027:Tobacco Mosaic Virus / Q000235:genetics / Q000276:immunology; D014611:Vaccination / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins