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In vivo effect of adhesion inhibitor heparin on Legionella pneumophila pathogenesis in a murine pneumonia model

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Abstract

Objective

To examine the effect of intratracheal heparin instillation on Legionella pneumophila-related acute lung injury (ALI) and systemic dissemination.

Design

Prospective, controlled experimental study.

Setting

University research laboratory.

Interventions

A/J mice received 5 μg of sulfated heparin intratracheally co-instilled with 106 or 108 colony-forming units (CFU) of a virulent isolate of L. pneumophila.

Measurements and results

ALI was assessed in control groups (PBS and PBS-heparin) and on days 1, 2 and 3 post-infection, in terms of the lung wet-to-dry (W/D) weight ratios and of lung endothelial permeability to radio-labeled albumin (Perm-I125). Lung bacterial loads were measured and systemic spread was assessed by blood and target organ culture. The alveolar inflammatory response was evaluated by measuring the cytokine levels (TNF-α, IFN-γ, IL-6 and IL-12p70) in bronchoalveolar lavage fluids (BALF). Co-instilled heparin improved mouse survival after the 108 CFU challenge (p < 0.01). On day 2, heparin co-instillation significantly reduced the W/D ratio and Perm-I125 (p < 0.01 and p < 0.001 respectively), improved lung bacterial clearance (p < 0.001), prevented systemic dissemination (blood, liver, spleen, kidneys and brain cultures, all p < 0.05) and significantly increased IFN-γ and IL-12p70 levels in BALF (p < 0.05).

Conclusions

Heparin co-instillation during intratracheal L. pneumophila challenge has a protective effect on the alveolar–capillary barrier and prevents bacterial dissemination. These results tend to confirm the competitive inhibition by heparin of L. pneumophila attachment to lung epithelium in vivo, and point to the possible involvement of a heparan-sulfate adhesin in L. pneumophila binding to pneumocytes.

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Acknowledgements

This study was supported by grants from Collège français des universitaires de Maladies Infectieuses et Tropicales (CMIT) and by Agence française de sécurité sanitaire de l’environnement et du travail (Afsset). The authors thank Dr. Bruno Cardinaud and Dr. Elizabeth Pradel for substantial assistance with work relevant to the study and David Young for editing the manuscript.

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Authors and Affiliations

  1. Université Lyon 1, INSERM U851, Centre National de référence des Légionelles, Faculté de Médecine Laennec, 7 rue Guillaume Paradin, 69372, Lyon cedex 08, France

    Florence Ader, Sophie Jarraud, Jerome Etienne & Christian Chidiac

  2. Centre de Biologie Est, Hospices Civils de Lyon, 69003, Lyon, France

    Sophie Jarraud & Jerome Etienne

  3. Laboratoire de Recherche en Pathologie Infectieuse, EA 2689, Faculté de Médecine de Lille, Place de Verdun, 59045, Lille cedex, France

    Florence Ader, Rozenn Le Berre, Rémi Fackeure, Nathalie Viget, Karine Faure, Eric Kipnis & Benoît Guery

  4. Service de Maladies Infectieuses et Tropicales, Hospices Civils de Lyon, Hôpital de La Croix-Rousse, 69004, Lyon, France

    Florence Ader & Christian Chidiac

  5. INSERM–Groupe AVENIR Pathogénie des Légionelles, Institut Pasteur Lille–IBL, rue du Pr Calmette, 59021, Lille cedex, France

    Dominique Raze & Franco Dante Menozzi

Authors
  1. Florence Ader
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  2. Rozenn Le Berre
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  3. Rémi Fackeure
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  4. Dominique Raze
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  5. Franco Dante Menozzi
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  6. Nathalie Viget
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  7. Karine Faure
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  9. Benoît Guery
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  10. Sophie Jarraud
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  11. Jerome Etienne
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  12. Christian Chidiac
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Corresponding author

Correspondence to Florence Ader.

Additional information

Franco Dante Menozzi has died since this study was conducted.

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Ader, F., Le Berre, R., Fackeure, R. et al. In vivo effect of adhesion inhibitor heparin on Legionella pneumophila pathogenesis in a murine pneumonia model. Intensive Care Med 34, 1511–1519 (2008). https://doi.org/10.1007/s00134-008-1063-2

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  • DOI: https://doi.org/10.1007/s00134-008-1063-2

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