Akos Heinemann, MD, Gunter J. Sturm, MD, Martina Ofner, BSc, Eva M

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Presentation transcript:

Stem cell factor stimulates the chemotaxis, integrin upregulation, and survival of human basophils Akos Heinemann, MD, Gunter J. Sturm, MD, Martina Ofner, BSc, Eva M. Sturm, MSc, Charlotte Weller, PhD, Bernhard A. Peskar, MD, Adele Hartnell, PhD Journal of Allergy and Clinical Immunology Volume 116, Issue 4, Pages 820-826 (October 2005) DOI: 10.1016/j.jaci.2005.06.008 Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 SCF enhances the migration of basophils. Basophils were mixed with vehicle or agonists (top wells) and were allowed to migrate toward agonists in the bottom wells. The amount of migrated cells was expressed relative to nonstimulated migration (chemotactic index). ∗P < .05, SCF versus vehicle (n = 6-8). PDG2, Prostaglandin D2. Journal of Allergy and Clinical Immunology 2005 116, 820-826DOI: (10.1016/j.jaci.2005.06.008) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 SCF enhances the chemoattractant-induced CD11b upregulation in basophils through c-kit tyrosine kinase. In A and B, PBMCs were pretreated with anti-human c-kit antibody (clone 47233, 20 μg/mL) or IgG1 and then mixed with vehicle or SCF (10 nM) and stimulated with eotaxin or MCP-2. In C, samples of whole blood were pretreated with the tyrosine kinase inhibitor STI-571 (1 μM) and then mixed with vehicle or SCF (10 nM) and stimulated with eotaxin. CD11b upregulation in basophils was quantified as a percentage of the baseline value. The P values show the levels of probability for differences between the indicated treatments (2-way ANOVA). ∗P < .05, vehicle versus SCF pretreatment (n = 6-8). Journal of Allergy and Clinical Immunology 2005 116, 820-826DOI: (10.1016/j.jaci.2005.06.008) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 Basophils express c-kit, which is upregulated by IL-3. In A, samples of whole blood were stained with the anti-c-kit antibody YB5.B8 (filled histograms) or the IgG1 control antibody (open histograms). The tracings are representative of 9 donors. In B and C, isolated basophils were cultured in the presence (bold histogram) or absence (filled histogram) of IL-3 (300 pM) for up to 48 hours, and cells were stained with the anti-human c-kit antibody (clone YB5.B8, bold and filled histograms) or IgG1 (open histogram). A representative staining and means ± SEM of 4 experiments are shown. ∗P < .05 versus baseline. Journal of Allergy and Clinical Immunology 2005 116, 820-826DOI: (10.1016/j.jaci.2005.06.008) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 PI-3 kinase mediates the SCF-induced enhancement of basophil CD11b upregulation and chemotaxis. In A, PBMCs were pretreated with the PI-3 kinase inhibitor LY-294002 (20 μM) or vehicle and then mixed with vehicle or SCF (10 nM) and were finally stimulated with eotaxin. CD11b upregulation in basophils was quantified as a percentage of the baseline value. In B, basophils pretreated with the PI-3 kinase inhibitor LY-294002 (20 μM) or vehicle and mixed with vehicle or SCF (10 nM, top wells) were allowed to migrate toward eotaxin or C5a (bottom wells). The amount of migrated cells was expressed relative to nonstimulated migration (chemotactic index). The P values denote significant overall differences between vehicle and pretreatment (2-way ANOVA). ∗P < .05, SCF versus its vehicle (n = 6-8). Journal of Allergy and Clinical Immunology 2005 116, 820-826DOI: (10.1016/j.jaci.2005.06.008) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions