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Infection and Immunity, June 2003, p. 3235-3239, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3235-3239.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
 

Protection against Shiga Toxin 1 Challenge by Immunization of Mice with Purified Mutant Shiga Toxin 1

Satoshi Ishikawa,¹ Kazuyoshi Kawahara,² Yutaka Kagami,² Yasunori Isshiki,^2, Aki Kaneko,^2, Hidenori Matsui,³ Nobuhiko Okada,¹ and Hirofumi Danbara^1*

Department of Microbiology, School of Pharmaceutical Sciences,¹ Kitasato Institute for Life Science, Kitasato University, Tokyo 108-8641,³ Department of Bacteriology, The Kitasato Institute, Tokyo 108-8642, Japan²

Received 13 November 2002/ Returned for modification 17 January 2003/ Accepted 24 February 2003

Shiga toxin 1 (Stx1) of enterohemorrhagic Escherichia coli O157:H7 was cloned, and four mutant Stx1s were constructed by site-directed mutagenesis with PCR. The wild-type and mutant Stx1s with amino acid replacements at positions 167 and 170 of the A subunit were purified by one-step affinity chromatography with commercially available Globotriose Fractogel, and the mutant Stxs were used for the immunization of mice. The mutant toxins were nontoxic to Vero cells in vitro and to mice in vivo and induced the immunoglobulin G antibody against the wild-type Stx1, which neutralized the cytotoxicity of Stx1. The induced antibody titers depended on the mutation at position 170 of the A subunit. The mice immunized with the mutant Stx1s were protected against a challenge of approximately 100 times the 50% lethal dose of the wild-type Stx1, suggesting that the mutant toxins are good candidates for toxoid vaccines for infection by Stx1-producing E. coli.

* Corresponding author. Mailing address: Department of Microbiology, School of Pharmaceutical Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan. Phone: 81-3-5791-6255. Fax: 81-3-3444-4831. E-mail: danbarah@pharm.kitasato-u.ac.jp.

Editor: A. D. O'Brien

Present address: Department of Microbiology, School of Pharmaceutical Sciences, Josai University, Sakado-shi, Saitama 350-0295, Japan.

Present address: National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan.

Infection and Immunity, June 2003, p. 3235-3239, Vol. 71, No. 6
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.6.3235-3239.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
 

Copyright © 2003 by the American Society for Microbiology. All rights reserved.