Expression of activated FcγRII discriminates between multiple granulocyte-priming phenotypes in peripheral blood of allergic asthmatic subjects Deon.

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Presentation transcript:

Expression of activated FcγRII discriminates between multiple granulocyte-priming phenotypes in peripheral blood of allergic asthmatic subjects Deon Kanters, BSc, Willem ten Hove, MSc, Bart Luijk, MD, PhD, Corneli van Aalst, BSc, René C. Schweizer, MD, PhD, Jan-Willem J. Lammers, MD, PhD, Hubert G.M. Leufkens, PharmD, PhD, Jan A.M. Raaijmakers, PhD, Madelon Bracke, PhD, Leo Koenderman, PhD Journal of Allergy and Clinical Immunology Volume 120, Issue 5, Pages 1073-1081 (November 2007) DOI: 10.1016/j.jaci.2007.06.021 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 MoPhab A17 specifically recognizes FcγRIIA (CD32) on the murine pre–B-cell line Ba/F3. Ba/F3 cells were stably transfected with FcγRIIA or FcαRI (CD89) as a control. Thereafter, the cells were double stained with MoPhab A17 (FITC labeled) and with either IV.3/CD32 (phycoerythrin [PE] labeled) or A59/CD89 (PE labeled). Fluorescence is depicted in arbitrary units (AU). Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 MoPhab A17 only recognizes FcγRIIA (CD32) when neutrophils are activated with fMLF. Median channel fluorescence is depicted in arbitrary units (AU). Whole blood was activated with different amounts of fMLF before erythrocytes were lysed, and the leukocytes were stained with MoPhab A17 or mAb IV.3 (CD32). Neutrophils were identified according to their specific side-scatter and forward-scatter signals. Fluorescence is depicted in arbitrary units (AU). Data shown are means of 3 different experiments. Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 In vivo priming of eosinophils (A, B, and D-F) and neutrophils (A and C-F) in asthmatic subjects compared with cells from control subjects. Priming was measured by means of determination of expression of MoPhab A17 (Fig 3, A-C), αm (CD11b; Fig 3, D and F), or L-selectin (CD62L; Fig 3, E and F). Statistics was performed with Mann-Whitney U tests. Median channel fluorescence is depicted in arbitrary units (AU). Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Integrin subunit αm/CD11b expression correlates positively with L-selectin/CD62L expression on eosinophils (A; n = 25) and neutrophils (B; n = 23) in peripheral blood of asthmatic subjects. Data regarding αm/CD11b expression levels are plotted against the L-selectin/CD62L expression values within the same patient. Median cell fluorescence (MCF) is presented in arbitrary units (AU). Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Differential MoPhab A17 expression before and after allergen challenge on eosinophils and neutrophils between dual (n = 10) and single (n = 7) responders. The differences in priming were analyzed with the Mann-Whitney U test. Differences between prechallenge and postchallenge MoPhab A17 expressions were analyzed with Wilcoxon signed-rank tests. Data are depicted as median channel fluorescence (MCF) and given in arbitrary units (AU). Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Differential regulation of responsiveness for fMLF in neutrophils and eosinophils isolated from peripheral blood of dual and single responders. Priming was measured on the basis of expression of MoPhab A17 on eosinophils and neutrophils of dual responders (A and B, respectively) and single responders (C and D, respectively) before and 6 hours after allergen challenge. Data were analyzed with Wilcoxon signed-rank tests. NS, Not significant; MCF, median channel fluorescence; AU, arbitrary units. Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 Low baseline (before allergen challenge) log2 PC20 methacholine values (<1.5 mg/mL) correlate with αm/CD11b expression on eosinophils (A) and neutrophils (B) isolated from peripheral blood of dual responders (n = 9). Data regarding αm/CD11b expression levels are plotted against the baseline PC20 methacholine in the same patient. Median channel fluorescence (MCF) is given in arbitrary units (AU). Journal of Allergy and Clinical Immunology 2007 120, 1073-1081DOI: (10.1016/j.jaci.2007.06.021) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions