Modulation of β1-integrins on hemopoietic progenitor cells after allergen challenge in asthmatic subjects Adriana E. Catalli, PhD, Jennifer V. Thomson,

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Presentation transcript:

Modulation of β1-integrins on hemopoietic progenitor cells after allergen challenge in asthmatic subjects Adriana E. Catalli, PhD, Jennifer V. Thomson, BSc, Irene M. Babirad, BSc, MyLinh Duong, MD, Tracey M. Doyle, BSc, Karen J. Howie, BSc, Paul Newbold, PhD, Richard I. Craggs, PhD, Martyn Foster, PhD, Gail M. Gauvreau, PhD, Paul M. O'Byrne, MD, Roma Sehmi, PhD Journal of Allergy and Clinical Immunology Volume 122, Issue 4, Pages 803-810 (October 2008) DOI: 10.1016/j.jaci.2008.07.021 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Detection of VLA-4 expression on BM-derived CD34+CD45+ cells by means of multigating flow cytometry. Plots represent BM, low-density, and nonadherent mononuclear cells stained with CD45–fluorescein isothiocyanate (FITC)/CD34–phycoerythrin (PE). Events in region R4 (true progenitors; identified as CD34high/CD45dull cells with low side scatter) were evaluated for SMFI and analyzed for specific staining with peridinin chlorophyll protein (PerCp)–linked VLA-4 (CD49d) antibody or isotype control. Data were collected with 99% confidence limits (set relative to 1% cells stained with isotype). SSC, Side scatter; FSC, forward scatter. Journal of Allergy and Clinical Immunology 2008 122, 803-810DOI: (10.1016/j.jaci.2008.07.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 CD34+CD45+ cell numbers at baseline and several time points after allergen challenge in BM (A; n = 12-13) and PB (B; n = 16 for all time points). Values were log10 transformed to fit a normal distribution before analysis and expressed as the number of CD34+CD45+ cells per 1 million white blood cells (WBC). Difference from baseline: ∗P < .05. Journal of Allergy and Clinical Immunology 2008 122, 803-810DOI: (10.1016/j.jaci.2008.07.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Allergen-induced changes in integrin expression on BM- and PB-derived CD34+CD45+ cells expressed as SMFI. VLA-4 expression decreased after challenge in BM (A; n = 9-15) and PB (B; n = 13-15). VLA-5 expression was unchanged in BM (C; n = 12-16) but decreased in PB (D; n = 16 for all time points). Mac-1 expression was unchanged (E and F; n = 16 for all time points). Difference from baseline: ∗P < .05; difference from BM baseline: †P < .05. Journal of Allergy and Clinical Immunology 2008 122, 803-810DOI: (10.1016/j.jaci.2008.07.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Adhesion of BM-derived CD34+CD45+ cells to BM components expressed as a percentage of total cells added. SDF-1–induced progenitor cell adhesion to fibronectin (A) was attenuated by anti-VLA-4 and anti-VLA-5 antibodies (n = 4 for unstimulated and n = 5 for all other conditions) and that to VCAM-1 (B) was attenuated by anti-VLA-4 antibodies (n = 5 for all conditions). Difference from SDF-1 baseline: ∗P < .05. Journal of Allergy and Clinical Immunology 2008 122, 803-810DOI: (10.1016/j.jaci.2008.07.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Effect of allergen challenge on BM-derived CD34+CD45+ cell adhesion to fibronectin. Unstimulated (n = 7) and SDF-1 (1 and 10 ng/mL; n = 9 and n = 8, respectively)–stimulated adhesion to fibronectin was reduced 24 hours after allergen challenge. Adhesion began to recover by 48 hours (inset), as determined from 1 individual's samples (expressed as mean of duplicate assessment). Difference from baseline: ∗P < .05. Journal of Allergy and Clinical Immunology 2008 122, 803-810DOI: (10.1016/j.jaci.2008.07.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions