Immunization with cDNA of a novel P-III type metalloproteinase from the rattlesnake Crotalus durissus durissus elicits antibodies which neutralize 69% of the hemorrhage induced by the whole venom.
Zinc-dependent metalloproteinases play a key role in the hemorrhage induced by viperid bite envenoming. In this work we report the cloning and sequencing of the cDNA from a novel P-III type metalloproteinase from Crotalus durissus durissus venom glands. The recombinant plasmid was used for DNA immunization in mice using accelerate DNA-coated microparticles with the Gene Gun system. The results showed that there is no significant difference in the efficiency of the immunization in mice when gold or tungsten microparticles were used. A pool of the sera from mice immunized with the metalloproteinase encoding DNA neutralized the hemorrhagic activity of C. d. durissus venom. The co-immunization with DNA encoding the metalloproteinase and a plasmid encoding the murine IL-2 increased the number of mice which show a specific antibody response towards C. d. durissus venom antigens. The neutralizing ability of the produced antibodies demonstrates that DNA immunization with tungsten microparticles may be used in strategies for antivenom production.
Azofeifa-Cordero
Gabriela
G
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Arce-Estrada
Viviana
V
Flores-Díaz
Marietta
M
Alape-Girón
Alberto
A
D000595:Amino Acid Sequence; D000818:Animals; D000997:Antivenins / Q000494:pharmacology*; D001483:Base Sequence; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors / Q000201:enzymology* / Q000633:toxicity; D018076:DNA, Complementary / Q000235:genetics / Q000276:immunology*; D006470:Hemorrhage / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control*; D045726:Metalloproteases / Q000145:classification / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017422:Sequence Analysis, DNA; D012867:Skin / Q000187:drug effects / Q000473:pathology
D000997:Antivenins; D003435:Crotalid Venoms; D018076:DNA, Complementary; D045726:Metalloproteases