Human TH2 cells respond to cysteinyl leukotrienes through selective expression of cysteinyl leukotriene receptor 1 Celine N. Parmentier, MSc, Elisabeth.

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Presentation transcript:

Human TH2 cells respond to cysteinyl leukotrienes through selective expression of cysteinyl leukotriene receptor 1 Celine N. Parmentier, MSc, Elisabeth Fuerst, PhD, Joanne McDonald, PhD, Holly Bowen, PhD, Tak H. Lee, MD, ScD, James E. Pease, PhD, Grzegorz Woszczek, MD, PhD, David J. Cousins, PhD Journal of Allergy and Clinical Immunology Volume 129, Issue 4, Pages 1136-1142 (April 2012) DOI: 10.1016/j.jaci.2012.01.057 Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 TH2 cells selectively express CYSLTR1. A, Intracellular cytokine staining of human TH1 and TH2 cells. Cells were either resting or activated for 4 hours with phorbol 12-myristate 13-acetate (5 ng/mL) and ionomycin (500 ng/mL). B and C, Real-time RT-PCR analysis of TH1 and TH2 cells for IFN-γ and IL-5 (Fig 1, B) and CYSLTR1 and CYSLTR2 (Fig 1, C). D, Time-course analysis of gene expression during TH1/TH2 differentiation by using real-time RT-PCR. Data shown are representative of 5 experiments with different donors except Fig 1, B, C, and D, which show means ± SEMs of 5 experiments. *P < .05, **P < .01, and ***P < .001, 2-way ANOVA with Bonferroni post test. A, Activated; R, resting. Journal of Allergy and Clinical Immunology 2012 129, 1136-1142DOI: (10.1016/j.jaci.2012.01.057) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 TH2 cells express functional CysLT1. A, Calcium flux in response to increasing concentrations of LTD4 in TH1 and TH2 cells. B, Calcium flux in TH2 cells in response to increasing concentrations of LTC4, LTD4, and LTE4. C, Inhibition of calcium flux in TH2 cells in response to 100 nmol/L LTD4 by 3 different CysLT1 antagonists: MK571, montelukast, and zafirlukast. Data are presented as mean ± SEM percentages of maximum response to LTD4 from 3 experiments with different donors. ***P < .001, 1-way ANOVA with Tukey post test. Journal of Allergy and Clinical Immunology 2012 129, 1136-1142DOI: (10.1016/j.jaci.2012.01.057) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 CysLT1 is partially Gi and Gq coupled in TH2 cells. A, Pertussis toxin (PTX)–mediated inhibition of calcium response to LTD4 in TH2 cells. ***P < .001, 2-way ANOVA with Bonferroni post test. B, LTD4-mediated inhibition of cAMP signaling in TH2 cells. Cells were treated with forskolin to induce cAMP, and increasing concentrations of LTD4 inhibited cAMP generation. C, The CysLT1 antagonist MK571 blocks the LTD4-mediated inhibition of cAMP response. Cells were treated with 100 nmol/L LTD4. D, Thapsigargin (THP)–mediated inhibition and EDTA-mediated partial inhibition of calcium flux response to LTD4 in TH2 cells. All data are expressed as mean ± SEM percentages of maximum response to LTD4 or forskolin from 3 experiments with different donors. Fig 3, B and D: **P < .01 and ***P < .001, 1-way ANOVA with Tukey post test. Journal of Allergy and Clinical Immunology 2012 129, 1136-1142DOI: (10.1016/j.jaci.2012.01.057) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 LTD4 is chemotactic for TH2 cells. Chemotactic agents were assessed in a Transwell-based system. Cells were allowed to migrate for 2 hours. Varying concentrations of LTD4 were used as shown. Control chemokines CXCL12 and CCL18 were used at 10 nmol/L concentration. Data shown are means ± SEMs of 3 independent experiments with different donors. ***P < .001 and *P < .05, 2-way ANOVA with Bonferroni post test. Journal of Allergy and Clinical Immunology 2012 129, 1136-1142DOI: (10.1016/j.jaci.2012.01.057) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions