This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kiyatkin, N. Articles by Simpson, L. L. Search for Related Content PubMed PubMed Citation Articles by Kiyatkin, N. Articles by Simpson, L. L.

 Previous Article  |  Next Article 

Infect. Immun., 11 1997, 4586-4591, Vol 65, No. 11
Copyright © 1997, American Society for Microbiology

Induction of an immune response by oral administration of recombinant botulinum toxin

N Kiyatkin, AB Maksymowych and LL Simpson
Department of Medicine, Jefferson Medical College, Philadelphia, Pennsylvania 19107, USA.

A gene encoding the full-size botulinum neurotoxin serotype C was reconstructed in vector pQE-30 and expressed at high levels in Escherichia coli. Three amino acid mutations (H229-->G, E230-->T, and H233-->N) were generated in the zinc-binding motif, resulting in complete detoxification of the modified recombinant holotoxin. The PCR- amplified wild-type light chain of botulinum neurotoxin serotype C was also expressed in E. coli and used as a control in all experiments. Modified recombinant holotoxin and light chain contained a histidine affinity tag at the amino terminus, which was used for detection and purification. Recombinant proteins were purified on nickel affinity resin and analyzed by Western blotting with the anti-histidine tag and anti-neurotoxin C antibodies. The results indicated that the 150-kDa molecule of modified recombinant holotoxin and the 50-kDa recombinant light chain were synthesized without degradation; however, E. coli did not provide for efficient nicking of modified recombinant toxin. Modified recombinant holotoxin was not toxic to mice, had no effect on nerve-evoked muscle twitch in vitro, and was not able to cleave syntaxin in crude synaptosome preparations. The recombinant light chain was also nontoxic in vivo, had no effect on evoked muscle twitch, but was able to cleave syntaxin. Modified recombinant neurotoxin and light chain were administered to animals either orally or subcutaneously. Both oral administration and subcutaneous administration of modified recombinant neurotoxin evoked high levels of serum antibodies and protective immunity. Oral administration of recombinant light chain evoked no systemic response, whereas subcutaneous administration evoked antibody production and immunity.

This article has been cited by other articles:

• Pier, C. L., Tepp, W. H., Bradshaw, M., Johnson, E. A., Barbieri, J. T., Baldwin, M. R. (2008). Recombinant Holotoxoid Vaccine against Botulism. Infect. Immun. 76: 437-442 [Abstract] [Full Text]  
• Dembek, Z. F., Smith, L. A., Rusnak, J. M. (2007). Botulism: Cause, Effects, Diagnosis, Clinical and Laboratory Identification, and Treatment Modalities. dmphp 1: 122-134 [Abstract] [Full Text]  
• Ravichandran, E., Al-Saleem, F. H., Ancharski, D. M., Elias, M. D., Singh, A. K., Shamim, M., Gong, Y., Simpson, L. L. (2007). Trivalent Vaccine against Botulinum Toxin Serotypes A, B, and E That Can Be Administered by the Mucosal Route. Infect. Immun. 75: 3043-3054 [Abstract] [Full Text]  
• Kobayashi, R., Kohda, T., Kataoka, K., Ihara, H., Kozaki, S., Pascual, D. W., Staats, H. F., Kiyono, H., McGhee, J. R., Fujihashi, K. (2005). A Novel Neurotoxoid Vaccine Prevents Mucosal Botulism. J. Immunol. 174: 2190-2195 [Abstract] [Full Text]  
• Arimitsu, H., Lee, J.-C., Sakaguchi, Y., Hayakawa, Y., Hayashi, M., Nakaura, M., Takai, H., Lin, S.-N., Mukamoto, M., Murphy, T., Oguma, K. (2004). Vaccination with Recombinant Whole Heavy Chain Fragments of Clostridium botulinum Type C and D Neurotoxins. CVI 11: 496-502 [Abstract] [Full Text]  
• Simpson, L. L., Maksymowych, A. B., Park, J.-B., Bora, R. S. (2004). The Role of the Interchain Disulfide Bond in Governing the Pharmacological Actions of Botulinum Toxin. J. Pharmacol. Exp. Ther. 308: 857-864 [Abstract] [Full Text]  
• Pless, D. D., Torres, E. R., Reinke, E. K., Bavari, S. (2001). High-Affinity, Protective Antibodies to the Binding Domain of Botulinum Neurotoxin Type A. Infect. Immun. 69: 570-574 [Abstract] [Full Text]  
• Maksymowych, A. B., Reinhard, M., Malizio, C. J., Goodnough, M. C., Johnson, E. A., Simpson, L. L. (1999). Pure Botulinum Neurotoxin Is Absorbed from the Stomach and Small Intestine and Produces Peripheral Neuromuscular Blockade. Infect. Immun. 67: 4708-4712 [Abstract] [Full Text]  
• Maksymowych, A. B., Simpson, L. L. (1998). Binding and Transcytosis of Botulinum Neurotoxin by Polarized Human Colon Carcinoma Cells. J. Biol. Chem. 273: 21950-21957 [Abstract] [Full Text]  
• Simpson, L. L., Maksymowych, A. B., Hao, S. (2001). The Role of Zinc Binding in the Biological Activity of Botulinum Toxin. J. Biol. Chem. 276: 27034-27041 [Abstract] [Full Text]