Generation of High-Titer Neutralizing Antibodies against Botulinum Toxins A, B, and E by DNA Electrotransfer
- 1 May 2009
- journal article
- Published by American Society for Microbiology in Infection and Immunity
- Vol. 77 (5), 2221-2229
- https://doi.org/10.1128/iai.01269-08
Abstract
Botulinum neurotoxins are known to be among the most toxic known substances. They produce severe paralysis by preventing the release of acetylcholine at the neuromuscular junction. Thus, new strategies for efficient production of safe and effective anti-botulinum neurotoxin antisera have been a high priority. Here we describe the use of DNA electrotransfer into the skeletal muscle to enhance antiserum titers against botulinum toxin serotypes A, B, and E in mice. We treated animals with codon-optimized plasmid DNA encoding the nontoxic but highly immunogenic C-terminal heavy chain fragment of the toxin. By employing both codon optimization and the electrotransfer procedure, the immune response and corresponding neutralizing antiserum titers were markedly increased. The cellular localization of the antigen and the immunization regimens were also shown to increase neutralizing titers to >100 IU/ml. This study demonstrates that DNA electrotransfer is an effective procedure for raising neutralizing antiserum titers to remarkably high levels.Keywords
This publication has 50 references indexed in Scilit:
- DNA vaccines: ready for prime time?Nature Reviews Genetics, 2008
- Antigenic and Immunogenic Study of Membrane-Proximal External Region-Grafted gp120 Antigens by a DNA Prime-Protein Boost Immunization StrategyJournal of Virology, 2007
- Identification of the protein receptor binding site of botulinum neurotoxins B and G proves the double-receptor conceptProceedings of the National Academy of Sciences of the United States of America, 2007
- The synaptic vesicle protein 2C mediates the uptake of botulinum neurotoxin A into phrenic nervesFEBS Letters, 2006
- Early events of electroporation‐mediated intramuscular DNA vaccination potentiate Th1‐directed immune responsesThe Journal of Gene Medicine, 2005
- Interaction between the two subdomains of the C‐terminal part of the botulinum neurotoxin A is essential for the generation of protective antibodiesFEBS Letters, 2004
- In vivo electroporation improves immune responses to DNA vaccination in sheepVaccine, 2004
- Codon optimization and mRNA amplification effectively enhances the immunogenicity of the hepatitis C virus nonstructural 3/4A geneGene Therapy, 2004
- DNA transfection of mononuclear cells in muscle tissueThe Journal of Gene Medicine, 2003
- DNA immunization with Onchocerca volvulus genes, Ov-tmy-1 and OvB20: serological and parasitological outcomes following intramuscular or GeneGun delivery in a mouse model of OnchocerciasisParasite Immunology, 2000