title: Neutralization of venom-induced hemorrhage by equine antibodies raised by immunization with a plasmid encoding a novel P-II metalloproteinase from the lancehead pitviper Bothrops asper.
abstract: In this work, the cDNA encoding a novel P-II type metalloproteinase from Bothrops asper venom glands was cloned, sequenced and used for DNA immunization of animals with accelerated DNA-coated tungsten microparticles and the helius Gene Gun system. Specific antibodies against B. asper venom antigens were induced in mice co-immunized with the plasmid encoding the P-II metalloproteinase together with an expression plasmid encoding the murine IL-2. Similarly, specific antibodies against B. asper venom antigens were also induced in a horse co-immunized with the plasmid encoding the P-II metalloproteinase, together with a plasmid encoding the equine IL-6. The equine antibodies induced by immunization with the P-II metalloproteinase encoding plasmid cross react with several proteins of B. asper, Crotalus durissus durissus, and Lachesis stenophrys venoms in western blot, demonstrating antigenic similarity between the cloned metalloproteinase and other metalloproteinases present in these venoms. Furthermore, the equine antibodies induced by immunization with the P-II metalloproteinase encoding plasmid completely neutralized the hemorrhagic activity of the whole B. asper venom and partially the hemorrhagic activity of C. durissus durissus venom. The neutralizing ability of the produced antibodies raises, for the first time, the possibility of developing therapeutic antivenoms in horses by DNA immunization using tungsten microparticles.
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: Viviana
authors.initials: V
authors.lastname: Arce-Estrada
authors.forename: Gabriela
authors.initials: G
authors.lastname: Azofeifa-Cordero
authors.forename: Ricardo
authors.initials: R
authors.lastname: Estrada
authors.forename: Alberto
authors.initials: A
authors.lastname: Alape-Girón
authors.forename: Marietta
authors.initials: M
authors.lastname: Flores-Díaz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000931:Antidotes / Q000494:pharmacology*; D000997:Antivenins / Q000494:pharmacology*; D001483:Base Sequence; D017837:Bothrops / Q000276:immunology*; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000276:immunology*; D018076:DNA, Complementary / Q000235:genetics; D006470:Hemorrhage / Q000276:immunology / Q000517:prevention & control*; D006736:Horses; D045726:Metalloproteases / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010957:Plasmids / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D019444:Vaccines, DNA / Q000276:immunology
chemical_list: D000931:Antidotes; D000997:Antivenins; D003435:Crotalid Venoms; D018076:DNA, Complementary; D019444:Vaccines, DNA; D045726:Metalloproteases