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Infection and Immunity, August 2001, p. 4823-4830, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4823-4830.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Major Outer Membrane Protein Omp25 of Brucella suis Is Involved in Inhibition of Tumor Necrosis Factor Alpha Production during Infection of Human Macrophages

Véronique Jubier-Maurin,1 Rose-Anne Boigegrain,1 Axel Cloeckaert,2 Antoine Gross,1 Maria-Teresa Alvarez-Martinez,1 Annie Terraza,1 Janny Liautard,1 Stephan Köhler,1 Bruno Rouot,1 Jacques Dornand,1 and Jean Pierre Liautard1,*

INSERM U431, Microbiologie et Pathologie Cellulaire Infectieuse, Université de Montpellier-II, 34095 Montpellier Cedex 05,1 and Laboratoire de Pathologie Infectieuse et Immunologie, I.N.R.A., 37380 Nouzilly,2 France

Received 28 February 2001/Returned for modification 12 April 2001/Accepted 24 May 2001

Brucella spp. can establish themselves and cause disease in humans and animals. The mechanisms by which Brucella spp. evade the antibacterial defenses of their host, however, remain largely unknown. We have previously reported that live brucellae failed to induce tumor necrosis factor alpha (TNF-alpha ) production upon human macrophage infection. This inhibition is associated with a nonidentified protein that is released into culture medium. Outer membrane proteins (OMPs) of gram-negative bacteria have been shown to modulate macrophage functions, including cytokine production. Thus, we have analyzed the effects of two major OMPs (Omp25 and Omp31) of Brucella suis 1330 (wild-type [WT] B. suis) on TNF-alpha production. For this purpose, omp25 and omp31 null mutants of B. suis (Delta omp25 B. suis and Delta omp31 B. suis, respectively) were constructed and analyzed for the ability to activate human macrophages to secrete TNF-alpha . We showed that, in contrast to WT B. suis or Delta omp31 B. suis, Delta omp25 B. suis induced TNF-alpha production when phagocytosed by human macrophages. The complementation of Delta omp25 B. suis with WT omp25 (Delta omp25-omp25 B. suis mutant) significantly reversed this effect: Delta omp25-omp25 B. suis-infected macrophages secreted significantly less TNF-alpha than did macrophages infected with the Delta omp25 B. suis mutant. Furthermore, pretreatment of WT B. suis with an anti-Omp25 monoclonal antibody directed against an epitope exposed at the surface of the bacteria resulted in substancial TNF-alpha production during macrophage infection. These observations demonstrated that Omp25 of B. suis is involved in the negative regulation of TNF-alpha production upon infection of human macrophages.

* Corresponding author. Mailing address: INSERM U431, IFR Eugène Bataillon, Université de Montpellier-II, 34095 Montpellier Cedex 05, France. Phone: (33) 467 14 32 09. Fax: (33) 467 14 33 38. E-mail: liautard{at}crit.univ-montp2.fr.

Infection and Immunity, August 2001, p. 4823-4830, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4823-4830.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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