A deep rough type structure in Bordetella bronchiseptica lipopolysaccharide modulates host immune responses

Authors

  • Federico Sisti,

    1. VacSal Laboratory, Biotechnology and Molecular Biology Institute, Department of Biological Sciences, Faculty of Sciences, National University of La Plata, National Council of Scientific and Technical Research (CONICET), Calles 47 y 115 (1900) La Plata
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    • These authors contributed equally to this work and share first author status.

  • Julieta Fernández,

    1. VacSal Laboratory, Biotechnology and Molecular Biology Institute, Department of Biological Sciences, Faculty of Sciences, National University of La Plata, National Council of Scientific and Technical Research (CONICET), Calles 47 y 115 (1900) La Plata
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    • These authors contributed equally to this work and share first author status.

  • Sarah C. Higgins,

    1. Immune Regulation Research Group, School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland
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  • Adriana Casabuono,

    1. Center for Carbohydrate Research, Department of Organic Chemistry, Faculty of Exact and Natural Science, (1428) University of Buenos Aires, Buenos Aires, Argentina
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  • Alicia Couto,

    1. Center for Carbohydrate Research, Department of Organic Chemistry, Faculty of Exact and Natural Science, (1428) University of Buenos Aires, Buenos Aires, Argentina
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  • Kingston H. G. Mills,

    1. Immune Regulation Research Group, School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland
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  • Daniela Hozbor

    1. VacSal Laboratory, Biotechnology and Molecular Biology Institute, Department of Biological Sciences, Faculty of Sciences, National University of La Plata, National Council of Scientific and Technical Research (CONICET), Calles 47 y 115 (1900) La Plata
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Daniela Hozbor, Laboratorio VacSal, Instituto de Biotecnología y Biología Molecular, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CCT La Plata CONICET, Calles 47 y 115 (1900) La Plata, Argentina. Tel: 54 221 422 9777; fax: 54 221 4229777; email address: hozbor@biol.unlp.edu.ar

ABSTRACT

The present authors have previously obtained the Bordetella bronchiseptica mutant BbLP39, which contains a deep-rough lipopolysaccharide (LPS) instead the wild type smooth LPS with O antigen. This mutant was found to be altered in the expression of some proteins and in its ability to colonize mouse lungs. Particularly, in BbLP39 the expression of pertactin is decreased. To differentiate the contribution of each bacterial component to the observed phenotype, here mice defective in the LPS sensing receptor TLR4 (TLR4-defective mice) were used. In contrast to wild-type mice, infection of TLR4-defective mice with BbLP39 resulted in lung infection, which persisted for more than 10 days post-challenge. Comparative analysis of the immune responses induced by purified mutant and wild type LPSs showed that the mutant LPS induced significantly higher degrees of expression of TNF-α and IL-10 mRNA than did the wild type. UV matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) mass spectrometry analysis revealed that both LPSs had the same penta-acylated lipid A structure. However, the lipid A from BbLP39 contained pyrophosphate instead of phosphate at position 1. This structural difference, in addition to the lack of O-antigen in BbLP39, may explain the functional differences between BbLP39 and wild type strains.

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