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Infection and Immunity, January 2008, p. 437-442, Vol. 76, No. 1
0019-9567/08/$08.00+0     doi:10.1128/IAI.00843-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Recombinant Holotoxoid Vaccine against Botulism{triangledown}

Christina L. Pier,1 William H. Tepp,1 Marite Bradshaw,1 Eric A. Johnson,1 Joseph T. Barbieri,2 and Michael R. Baldwin2*

Food Research Institute, University of Wisconsin, Madison, Wisconsin,1 Department of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, Wisconsin2

Received 19 June 2007/ Returned for modification 21 September 2007/ Accepted 10 October 2007

The botulinum neurotoxins (BoNT) are the most toxic proteins for humans and designated "Category A Select Agents." The current vaccine against botulism is in limited supply, and there is a need to develop new vaccine strategies. A recombinant BoNT/A toxoid was produced in Clostridium botulinum that contained a double amino acid substitution, R363A Y365F (termed BoNT/ARYM). BoNT/ARYM was noncatalytic for SNAP25 and nontoxic for mice. Immunization with BoNT/ARYM protected mice from challenge at levels that were similar to chemically inactivated BoNT/A toxoid. BoNT/ARYM elicited an immune response against the light-chain and heavy-chain components of the toxin. Neutralizing anti-BoNT/ARYM sera blocked BoNT toxicity in primary cortical neurons and blocked ganglioside binding by the heavy chain. BoNT/ARYM represents a viable vaccine candidate for a holotoxoid against botulism.

* Corresponding author. Mailing address: Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226. Phone: (414) 456-8419. Fax: (414) 456-6535. E-mail: mbaldwin{at}mcw.edu

{triangledown} Published ahead of print on 29 October 2007.

Editor: V. J. DiRita

Infection and Immunity, January 2008, p. 437-442, Vol. 76, No. 1
0019-9567/08/$08.00+0     doi:10.1128/IAI.00843-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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