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Clinical and Diagnostic Laboratory Immunology, May 2004, p. 496-502, Vol. 11, No. 3
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.3.496-502.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Vaccination with Recombinant Whole Heavy Chain Fragments of Clostridium botulinum Type C and D Neurotoxins

Hideyuki Arimitsu,^1, Jae-Chul Lee,¹ Yoshihiko Sakaguchi,¹ Yuji Hayakawa,² Michiko Hayashi,² Miki Nakaura,² Hikaru Takai,² Song-Nan Lin,¹ Masafumi Mukamoto,³ Tom Murphy,⁴ and Keiji Oguma^1*

Department of Bacteriology, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558,¹ Nanbu Livestock Hygiene Service Center, Kanazawa, Ishikawa 920-3101,² Department of Veterinary Epidemiology, University of Osaka Prefecture, Sakai, Osaka 599-8531, Japan,³ National Water Research Institute, Burlington, Ontario L7R 4A6, Canada⁴

Received 15 November 2003/ Returned for modification 22 December 2003/ Accepted 28 January 2004

Mice and ducks were subcutaneously immunized with recombinant whole heavy (H) chains of Clostridium botulinum type C and D neurotoxins, which were expressed as glutathione S-transferase fusion proteins. In the case of mice, it was confirmed that two immunizations with type C- and D-H chains, 10 µg each time, significantly increased the specific antibodies against 100-kDa H chains of type C and D neurotoxins in an immunoblot analysis and an enzyme-linked immunosorbent assay, respectively. The mice immunized with type C- and D-H chains showed no symptoms of botulism when they were challenged with C- and D-16 S toxins at doses, given intraperitoneally, of up to 10⁵ and 10⁶ minmum lethal doses (MLD), respectively, per mouse. Ducks were immunized with a total of 100 µg of type C-H chain. The ducks also developed specific antibodies to the type C-H chain and showed significant protection against a challenge with 10³ duck MLD of C-16 S toxin given intravenously. These results indicate that recombinant whole H chains can be used as an effective and safe vaccine for type C and D botulism in domestic animals.

Present address: Department of Microbiology, Fujita Health University, Toyoake, Aichi 470-1192, Japan.