Retinoic acid modulates IL-5 receptor expression and selectively inhibits eosinophil- basophil differentiation of hemopoietic progenitor cells John W.

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Presentation transcript:

Retinoic acid modulates IL-5 receptor expression and selectively inhibits eosinophil- basophil differentiation of hemopoietic progenitor cells John W. Upham, MD, PhDa*, Roma Sehmi, PhDb, Lisa M. Hayes, BScb, Karen Howie, b, Joachim Lundahl, MDb, Judah A. Denburg, MDb Journal of Allergy and Clinical Immunology Volume 109, Issue 2, Pages 307-313 (February 2002) DOI: 10.1067/mai.2002.121527 Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 1 ATRA inhibits eosinophil-basophil, but not granulocyte-macrophage, CFUs. Normal bone marrow was obtained from nonatopic subjects undergoing cardiac surgery. Low-density, nonadherent bone marrow cells were cultured in conventional semisolid colony-forming assays and supplemented with conditioned medium in the presence or absence of ATRA 10−6 mol/L. CFUs were identified and enumerated on day 14 by means of inverted light microscopy and classified as either eosinophil-basophil CFUs or as granulocyte-macrophage CFUs by using standard morphologic criteria.3 Data are expressed as mean of 6 experiments. *P < .05. Journal of Allergy and Clinical Immunology 2002 109, 307-313DOI: (10.1067/mai.2002.121527) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 2 ATRA inhibits surface expression of IL-5Rα but not GM-CSFRα. Cord blood CD34+ cells were isolated by means of positive immunomagnetic selection with MACS beads and studied after 7 days of liquid culture in the presence of 1.0 ng/mL IL-3, 1.0 ng/mL IL-5, and 10 ng/mL GM-CSF. Cells were stained with saturating amounts of either biotin-conjugated anti-IL-5Rα, anti-GM-CSFRα (black line) , or IgG1 isotype control (red line) antibodies, followed by streptavidin CyChrome. FACS analysis gates were set to exclude dead cells, and at least 30,000 events were acquired for each sample. Histograms of IL-5Rα expression (A , B , and C ) and GM-CSFRα expression (D , E , and F ) from a representative experiment are shown. RA , All-trans retinoic acid. Journal of Allergy and Clinical Immunology 2002 109, 307-313DOI: (10.1067/mai.2002.121527) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 3 ATRA inhibits surface expression of IL-5Rα in a dosedependent manner. Group data are shown (n = 5). ATRA significantly reduced both the proportion of cells expressing IL-5Rα (upper panel) and the intensity of receptor expression (lower panel) . Fluorescence intensity of cytokine receptor staining was determined relative to fluorescence of the isotype control, indicated in the lower panel by the dashed line , and arbitrarily set at 100. *P < .05. Journal of Allergy and Clinical Immunology 2002 109, 307-313DOI: (10.1067/mai.2002.121527) Copyright © 2002 Mosby, Inc. Terms and Conditions

Fig. 4 ATRA has no effect on surface expression of GM-CSFRα. Group data are shown, as in Fig 3. Data are shown only for 10−6 mol/L ATRA; 10−7 and 10−8 mol/L ATRA likewise had no effect on GM-CSFRα expression. The upper panel displays the proportion of cells expressing GM-CSFRα, whereas the intensity of receptor expression is shown in the lower panel . Fluorescence intensity of cytokine receptor staining was determined relative to fluorescence of the isotype control, indicated in the lower panel by the dashed line , and arbitrarily set at 100. Journal of Allergy and Clinical Immunology 2002 109, 307-313DOI: (10.1067/mai.2002.121527) Copyright © 2002 Mosby, Inc. Terms and Conditions