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Infection and Immunity, October 2007, p. 4867-4874, Vol. 75, No. 10
0019-9567/07/$08.00+0     doi:10.1128/IAI.00439-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

BvrR/BvrS-Controlled Outer Membrane Proteins Omp3a and Omp3b Are Not Essential for Brucella abortus Virulence{triangledown}

Lorea Manterola,1,{dagger} Caterina Guzmán-Verri,2* Esteban Chaves-Olarte,2,3 Elías Barquero-Calvo,2 María-Jesús de Miguel,4 Ignacio Moriyón,1 María-Jesús Grilló,5 Ignacio López-Goñi,1 and Edgardo Moreno2

Departamento de Microbiología y Parasitología, Universidad de Navarra, 31008 Pamplona, Spain,1 Programa de Investigación en Enfermedades Tropicales (PIET), Escuela de Medicina Veterinaria, Universidad Nacional, 304-3000 Heredia, Costa Rica,2 Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, 1000 San José, Costa Rica,3 Centro de Investigación y Tecnología Agroalimentaria (CITA), Sanidad Animal, Gobierno de Aragón, Zaragoza, Spain,4 Instituto de Agrobiotecnología, CSIC-UPNA-Gobierno de Navarra, Pamplona, Spain5

Received 26 March 2007/ Returned for modification 6 May 2007/ Accepted 17 July 2007

The Brucella abortus two-component regulatory system BvrR/BvrS controls the expression of outer membrane proteins (Omp) Omp3a (Omp25) and Omp3b (Omp22). Disruption of bvrS or bvrR generates avirulent mutants with altered cell permeability, higher sensitivity to microbicidal peptides, and complement. Consequently, the role of Omp3a and Omp3b in virulence was examined. Similar to bvrS or bvrR mutants, omp3a and omp3b mutants displayed increased attachment to cells, indicating surface alterations. However, they showed unaltered permeability; normal expression of Omp10, Omp16, Omp19, Omp2b, and Omp1; native hapten polysaccharide; and lipopolysaccharide and were resistant to complement and polymyxin B at ranges similar to those of the wild-type (WT) counterpart. Likewise, omp3a and omp3b mutants were able to replicate in murine macrophages and in HeLa cells, were resistant to the killing action of human neutrophils, and persisted in mice, like the WT strain. Murine macrophages infected with the omp3a mutant generated slightly higher levels of tumor necrosis factor alpha than the WT, whereas the bvrS mutant induced lower levels of this cytokine. Since the absence of Omp3a or Omp3b does not result in attenuation, it can be concluded that BvrR/BvrS influences additional Brucella properties involved in virulence. Our results are discussed in the light of previous works suggesting that disruption of omp3a generates attenuated Brucella strains, and we speculate on the role of group 3 Omps.

* Corresponding author. Mailing address: Programa de Investigación en Enfermedades Tropicales (PIET), Escuela de Medicina Veterinaria, Universidad Nacional, 304-3000 Heredia, Costa Rica. Phone: (506) 238-0761. Fax: (506) 238-1298. E-mail: catguz{at}medvet.una.ac.cr

{triangledown} Published ahead of print on 30 July 2007.

Editor: J. B. Bliska

{dagger} Present address: Unidad de Proteómica CIC bioGUNE, Parque Tecnológico de Bizkaia, Ed. 801-A, 48160 Derio, Bizkaia, Spain.

Infection and Immunity, October 2007, p. 4867-4874, Vol. 75, No. 10
0019-9567/07/$08.00+0     doi:10.1128/IAI.00439-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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