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Infection and Immunity, January 2003, p. 30-39, Vol. 71, No. 1
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.1.30-39.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Oral Immunization with ATP-Dependent Protease-Deficient Mutants Protects Mice against Subsequent Oral Challenge with Virulent Salmonella enterica Serovar Typhimurium

Hidenori Matsui,1,2* Masato Suzuki,3 Yasunori Isshiki,2,{dagger} Chie Kodama,1 Masahiro Eguchi,1,2 Yuji Kikuchi,1,2 Kenji Motokawa,4 Akiko Takaya,3 Toshifumi Tomoyasu,3 and Tomoko Yamamoto3

Laboratory of Immunoregulation, Department of Infection Control and Immunology, Kitasato Institute for Life Sciences, Kitasato University, Minato-ku, Tokyo 108-8641,1 Center for Basic Research, The Kitasato Institute, Minato-ku, Tokyo 108-8642,2 Department of Microbiology and Molecular Genetics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 263-8522,3 Division of Research and Development, Research Center for Biologicals, The Kitasato Institute, Kitamoto, Saitama 364-0026, Japan4

Received 29 April 2002/ Returned for modification 15 July 2002/ Accepted 15 October 2002

We evaluated the efficacy of mutants with a deletion of the stress response protease gene as candidates for live oral vaccine strains against Salmonella infection through infection studies with mice by using a Salmonella enterica serovar Typhimurium mutant with a disruption of the ClpXP or Lon protease. In vitro, the ClpXP protease regulates flagellum synthesis and the ClpXP-deficient mutant strain exhibits hyperflagellated bacterial cells (T. Tomoyasu et al., J. Bacteriol. 184:645-653, 2002). On the other hand, the Lon protease negatively regulates the efficacy of invading epithelial cells and the expression of invasion genes (A. Takaya et al., J. Bacteriol. 184:224-232, 2002). When 5-week-old BALB/c mice were orally administered 5 x 108 CFU of the ClpXP- or Lon-deficient strain, bacteria were detected with 103 to 104 CFU in the spleen, mesenteric lymph nodes, Peyer's patches, and cecum 1 week after inoculation and the bacteria then decreased gradually in each tissue. Significant increases of lipopolysaccharide-specific immunoglobulin G (IgG) and secretory IgA were detected at week 4 and maintained until at least week 12 after inoculation in serum and bile, respectively. Immunization with the ClpXP- or Lon-deficient strain protected mice against oral challenge with the serovar Typhimurium virulent strain. Both the challenged virulent and immunized avirulent salmonellae were completely cleared from the spleen, mesenteric lymph nodes, Peyer's patches, and even cecum 5 days after the challenge. These data indicate that Salmonella with a disruption of the ATP-dependent protease ClpXP or Lon can be useful in developing a live vaccine strain.

* Corresponding author. Mailing address: Laboratory of Immunoregulation, Department of Infection Control and Immunology, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan. Phone and fax: 81-3-5791-6267.E-mail: hmatsui{at}lisci.kitasato-u.ac.jp.

Editor: B. B. Finlay

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

Infection and Immunity, January 2003, p. 30-39, Vol. 71, No. 1
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.1.30-39.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sano, G.-i., Takada, Y., Goto, S., Maruyama, K., Shindo, Y., Oka, K., Matsui, H., Matsuo, K. (2007). Flagella Facilitate Escape of Salmonella from Oncotic Macrophages. J. Bacteriol. 189: 8224-8232 [Abstract] [Full Text]  
  • Matsui, H., Eguchi, M., Ohsumi, K., Nakamura, A., Isshiki, Y., Sekiya, K., Kikuchi, Y., Nagamitsu, T., Masuma, R., Sunazuka, T., Omura, S. (2005). Azithromycin Inhibits the Formation of Flagellar Filaments without Suppressing Flagellin Synthesis in Salmonella enterica Serovar Typhimurium. Antimicrob. Agents Chemother. 49: 3396-3403 [Abstract] [Full Text]  
  • Kodama, C., Matsui, H. (2004). Salmonella Flagellin Is Not a Dominant Protective Antigen in Oral Immunization with Attenuated Live Vaccine Strains. Infect. Immun. 72: 2449-2451 [Abstract] [Full Text]  


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