Simultaneous GeneGun immunisation with plasmids encoding antigen and GM-CSF: significant enhancement of murine antivenom IgG1 titres.
GeneGun DNA immunisation is a potent means of inducing antibody-dominant immune responses that we are exploiting to generate venom toxin-specific antibodies to improve the therapy of systemic envenoming by snakes. Here, we report that mice immunised with DNA encoding the carboxyl domain (JD9) of a haemorrhagic Zn metalloprotease (Jararhagin) in venom of the South American pit viper, Bothrops jararaca, and a plasmid expressing murine cytokine granulocyte/macrophage-colony stimulating factor (GM-CSF) raised significantly higher antigen-specific IgG1 titres than mice immunised with JD9 DNA alone. Serological responses to GeneGun JD9 DNA immunisation were shown to be dominated by IgG1, an IgG subclass associated with T lymphocyte helper 2 (Th2) immune responses. Further significant enhancement of JD9-specific IgG1 titres was achieved by increasing the number of immunisations. This report illustrates that DNA immunisation protocols to achieve high-titre, venom toxin-specific antibody production are well advanced and encourage the development of a DNA-based approach to antivenom production.
Harrison
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Alistair Reid Venom Research Unit, Division of Molecular Biology and Immunology, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UK. robharr@liverpool.ac.uk
Richards
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Laing
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Theakston
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D000818:Animals; D000941:Antigens / Q000235:genetics; D000997:Antivenins / Q000097:blood; D019470:Biolistics; D003435:Crotalid Venoms / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000235:genetics*; D000097146:Bothrops jararaca Venom
D000941:Antigens; D000997:Antivenins; D003435:Crotalid Venoms; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007074:Immunoglobulin G; D008666:Metalloendopeptidases; D019444:Vaccines, DNA; D000097146:Bothrops jararaca Venom