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Cysteinyl leukotrienes acting via granule membrane-expressed receptors elicit secretion from within cell-free human eosinophil granules  Josiane S. Neves,

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Presentation on theme: "Cysteinyl leukotrienes acting via granule membrane-expressed receptors elicit secretion from within cell-free human eosinophil granules  Josiane S. Neves,"— Presentation transcript:

1 Cysteinyl leukotrienes acting via granule membrane-expressed receptors elicit secretion from within cell-free human eosinophil granules  Josiane S. Neves, PhD, Amy L. Radke, BS, Peter F. Weller, MD  Journal of Allergy and Clinical Immunology  Volume 125, Issue 2, Pages (February 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Isolated granules expressed extracellular domains of cysLT1R (A) and cysLT2R (B) but not the carboxy-terminal intracellular domain of CysLT1R (C). Shaded histograms represent staining with control antibody (Ab). Solid and dashed lines represent staining with polyclonal specific antibodies (pAb) and anti-cysLTR pAbs neutralized by absorption with their respective immunogen peptide. Data are from 1 experiment, representative of 3. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Isolated granules were stimulated with different concentrations ( nmol/L) of LTC4(A), LTD4(B), and LTE4(C), and ECP levels were measured in the supernatants. Data are shown for 1 experiment, representative of 3. +, Significantly increased ECP release (P < .05) compared with nonstimulated granules. M, mol/L. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 MRS 2395, a selective P2Y12R antagonist, dose-dependently inhibited the ECP release induced by LTC4 30 nmol/L (A), LTD4 0.3 nmol/L (B) and 300 nmol/L (C), and LTE4 30 nmol/L (D). + and ∗, P <.05 for ECP released compared with nonstimulated and leukotriene-stimulated granules, respectively. Eosinophil (E) and isolated granule surface membranes (F) expressed the P2Y12R. Shaded histograms and solid lines represent staining with control antibody and with an anti-P2Y12R specific polyclonal antibody (pAb), respectively. G, The expression of the P2Y12R was confirmed on eosinophil and isolated granules by Western blots. Specificity of immunodetection was confirmed by neutralization with the anti-P2Y12R pAb by absorption with its respective immunogen peptide. All data are shown for 1 experiment, representative of 3. Eos, Eosinophils; Gran, granules; M, mol/L. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Montelukast dose-dependently inhibited ECP secretion induced by LTC4 30 nmol/L (A), LTD4 0.3 nmol/L (B) and 300 nmol/L (C), and LTE4 30 nmol/L (D). Data are shown for 1 experiment, representative of 3. + and ∗, P <.05 for ECP released compared with nonstimulated granules and leukotriene-stimulated granules, respectively. μ, mol/L. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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