[en] Live attenuated vaccines (LAVs) whose virulence would be controlled at the tissue level could be a crucial tool to effectively fight intracellular bacterial pathogens, because they would optimize the induction of protective immune memory while avoiding the long-term persistence of vaccine strains in the host. Rational development of these new LAVs implies developing an exhaustive map of the bacterial virulence genes according to the host organs implicated. We report here the use of transposon sequencing to compare the bacterial genes involved in the multiplication of Brucella melitensis, a major causative agent of brucellosis, in the lungs and spleens of C57BL/6 infected mice. We found 257 and 135 genes predicted to be essential for B. melitensis multiplication in the spleen and lung, respectively, with 87 genes common to both organs. We selected genes whose deletion is predicted to produce moderate or severe attenuation in the spleen, the main known reservoir of Brucella, and compared deletion mutants for these genes for their ability to protect mice against challenge with a virulent strain of B. melitensis. The protective efficacy of a deletion mutant for the plsC gene, implicated in phospholipid biosynthesis, is similar to that of the reference Rev.1 vaccine but with a shorter persistence in the spleen. Our results demonstrate that B. melitensis faces different selective pressures depending on the organ and underscore the effectiveness of functional genome mapping for the design of new safer LAV candidates.
Disciplines :
Microbiology
Author, co-author :
Barbieux, Emeline; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium ; Laboratoire de Parasitologie, ULB Center for Research in Immunology (U-CRI), Université Libre de Bruxelles, Gosselies, Belgium
Potemberg, Georges ; Centre Hospitalier Universitaire de Liège - CHU > > Département de Physique Médicale ; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium
Stubbe, François-Xavier; Unité de recherche en physiologie moléculaire (URPhyM), Laboratoire de Génétique moléculaire (GéMo), University of Namur, Namur, Belgium
Fraikin, Audrey; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium
Poncin, Katy; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium
Reboul, Angeline; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium
Rouma, Thomas; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium ; Laboratoire de Parasitologie, ULB Center for Research in Immunology (U-CRI), Université Libre de Bruxelles, Gosselies, Belgium
Zúñiga-Ripa, Amaia; Departamento de Microbiología y Parasitología - IDISNA, Universidad de Navarra, Pamplona, Spain
De Bolle, Xavier; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium
Muraille, Eric ; Unité de Recherche en Biologie des Microorganismes (URBM), Laboratoire d’Immunologie et de Microbiologie, NARILIS, University of Namur, Namur, Belgium ; Laboratoire de Parasitologie, ULB Center for Research in Immunology (U-CRI), Université Libre de Bruxelles, Gosselies, Belgium
Language :
English
Title :
Genome-wide analysis of Brucella melitensis growth in spleen of infected mice allows rational selection of new vaccine candidates
This work was supported by grants from the Fonds National de la Recherche Scientifique (FNRS) (CDR J.0120.18 and CDR J.0157.20 to E. M. and PDR T.0058.20 to X.D.B., Belgium). E.M. is a Senior Research Associate from the FRS-FNRS (Belgium). E.B., G.P. and F-X.S. hold FRIA PhD grants from the FRS-FNRS (Belgium). The funders played no role in study design, data collection, analysis and interpretation of data, or the writing of this manuscript. We thank K. Willemart, F. Tilquin, E. Carlier and M. Waroquier for their technical support.
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