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Animal shed Bacillus licheniformis spores possess allergy-protective as well as inflammatory properties  Kay Vogel, MSc, Nicole Blümer, PhD, Melanie Korthals,

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Presentation on theme: "Animal shed Bacillus licheniformis spores possess allergy-protective as well as inflammatory properties  Kay Vogel, MSc, Nicole Blümer, PhD, Melanie Korthals,"— Presentation transcript:

1 Animal shed Bacillus licheniformis spores possess allergy-protective as well as inflammatory properties  Kay Vogel, MSc, Nicole Blümer, PhD, Melanie Korthals, MSc, Jessica Mittelstädt, PhD, Holger Garn, PhD, Markus Ege, MD, Erika von Mutius, MD, Sören Gatermann, MD, Albrecht Bufe, MD, Torsten Goldmann, PhD, Karin Schwaiger, MD, Harald Renz, MD, Sven Brandau, PhD, Johann Bauer, PhD, Holger Heine, PhD, Otto Holst, PhD  Journal of Allergy and Clinical Immunology  Volume 122, Issue 2, Pages e8 (August 2008) DOI: /j.jaci Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Stimulation of PBMCs by Blspo, hiBlspo, and vegBl. Data show means ± SEMs of TNF-α measurement in culture supernatant after dose-dependent (MOI) PBMC stimulation for 24 hours of 3 independent experiments from different healthy donors. ∗P < .05; ∗∗P < .01. n.s., Not significant; unst., unstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Stimulation of moDCs by Bacillus licheniformis 467 (MOI = 1) for 24 hours (A) before mixed leukocyte reaction (B). Cells were isolated from peripheral blood of healthy donors. Results of cytokine measurements are expressed as the mean of 3 independent experiments ± SEM. BCG, Mycobacterium bovis; DC, unstimulated DCs plus leukocytes; LPS, LPS 10 ng/mL; TC, leukocytes alone; unst., unstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Effect of intranasal treatment with Bacillus licheniformis 467 on the airway inflammation of ovalbumin-sensitized and challenged mice (n = 8). Mice BAL fluid was assessed for numbers of total eosinophils (A), neutrophils (B), lymphocytes (C), and macrophages (D). Depicted are means ± SEMs; significant differences between sham-treated (PBS) mice and bacteria treated mice are indicated by ∗P < .05; ∗∗P < .01; ∗∗∗P < # indicates significant differences between vegBI-treated mice and control medium–treated mice. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig. 4 Prevention of goblet cell hyperplasia (A) and fusion of mononuclear cells (B) by Bacillus licheniformis 467 treatment. Shown are representative micrographs of mice airways after periodic acid-Schiff staining. A, I, sham-treated mice; II, Blspo-treated mice; III, hiBlspo-treated mice; IV, vegBl-treated mice; V, control medium–treated mice. B, Formation of multinucleated giant cells shown is representative of all B licheniformis–treated groups. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 BAL cytokines IFN-Y (A), IL-ID (B), and IL-5 (C) measured after B licheniformis 467 treatment of mice sensitized and challenged to ovalbumin (n = 8). The determination of cytokine levels was performed by ELISA of BAL fluid. Shown are results as means ± SEMs for each treated group. Significant differences between sensitized PBS-treated and bacteria-treated mice are indicated by ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Schematic protocol of the experimental design for mouse experiments
Schematic protocol of the experimental design for mouse experiments. Female BALB/c mice were sensitized at days 0, 7, and 14 by ovalbumin (OVA) intraperitoneal (i.p.) injections followed by OVA aerosol exposure at days 19, 20, and 21. Twelve days before the first OVA injection and during the whole sensitization period, mice received B licheniformis 467 spores, heat-inactivated B licheniformis 467 spores, or vegetative B licheniformis 467 intranasally every second day. Mice were analyzed at days 22 and 23 for airway inflammation and lung function. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Transmission electron micrograph of B licheniformis 467
Transmission electron micrograph of B licheniformis 467. The vegetative bacterium, obtained from the log-phase of growth, is shown in longitudinal (A) and cross (B) sections. C, The dormant spore protected by several layers of highly cross-linked proteins and peptidoglycan. Bars indicate 0.5 μm. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Analyses of lung function by head-out body plethysmography
Analyses of lung function by head-out body plethysmography. Mice sensitized and challenged to ovalbumin, during and after bacterial treatment with B licheniformis 467 as the indicated forms. Methacholine concentrations that caused 50% reduction in baseline midexpiratory airflow (MCH50) are displayed. No treatment was able to achieve significant increase in airway hyperreactivity. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Lung tissue of B licheniformis 467 spore–treated mice after Wirtz-Conklin staining phagocytosed by alveolar macrophages. Staining was performed by using paraffin-embedded lung tissue 4 days after the last spore treatment. Left upper corner shows B licheniformis spores on an object slide. Magnification ×1000. Similar results were obtained by staining lung cytospins of Blspo and hiBlspo. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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