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Infection and Immunity, August 2001, p. 4938-4943, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4938-4943.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Stress-Induced ClpP Serine Protease of Listeria monocytogenes Is Essential for Induction of Listeriolysin O-Dependent Protective Immunity

Olivier Gaillot,1 Søren Bregenholt,2,3 Francis Jaubert,4 James P. Di Santo,2,3 and Patrick Berche1,*

INSERM U411,1 INSERM U429,2 and Service d'Anatomie Pathologique,4 Centre Hospitalo-Universitaire Necker-Enfants Malades, and Unité des Cytokines et Développement Lymphoïde, Institut Pasteur,3 Paris, France

Received 21 November 2000/Returned for modification 22 February 2001/Accepted 1 May 2001

The stress-induced protease ClpP is required for virulence of the facultative intracellular pathogen Listeria monocytogenes. We previously found that in the absence of ClpP, the virulence of this pathogen was strongly reduced, mainly due to the decreased production of functional listeriolysin O (LLO), a major immunodominant virulence factor promoting intracellular growth. In this work, a clpP deletion mutant of L. monocytogenes was used to study the generation of anti-Listeria protective immunity. We found that ClpP is required for the intracellular growth of L. monocytogenes in resident macrophages in vivo. Mice infected with doses as high as 106 clpP mutant bacteria were not protected against a lethal challenge of wild-type bacteria and did not develop any detectable LLO-specific cytolytic T cells or antibodies, suggesting that the amount of LLO produced in infected mice under these conditions was too low to induce a specific immune response. However, in contrast to the results obtained with a mutant with a disrupted hly gene, this lack of protection was overcome by inoculation of very high infecting doses of clpP mutant bacteria (5 × 108), thus producing sufficient amounts of LLO to stimulate anti-Listeria immunity. The role of ClpP was confirmed by showing that anti-Listeria immunity was restored in mice infected with a clpP-complemented mutant. These results indicate that the stress-induced serine protease ClpP is a potential target for modulating the presentation of protective antigens such as LLO and thereby the immune response against L. monocytogenes.

* Corresponding author. Mailing address: INSERM U411, Faculté de Médecine Necker-Enfants Malades, 156 Rue de Vaugirard, 75730 Paris Cedex 15, France. Phone: 33 1 40 61 53 71. Fax: 33 1 40 61 55 92. E-mail: berche{at}necker.fr.

Infection and Immunity, August 2001, p. 4938-4943, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4938-4943.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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