Thymic stromal lymphopoietin as a mediator of crosstalk between bronchial smooth muscles and mast cells  Zoulfia Allakhverdi, PhD, Michael R. Comeau,

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Thymic stromal lymphopoietin as a mediator of crosstalk between bronchial smooth muscles and mast cells  Zoulfia Allakhverdi, PhD, Michael R. Comeau, BSc, Heidi K. Jessup, BSc, Guy Delespesse, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 4, Pages 958-960.e2 (April 2009) DOI: 10.1016/j.jaci.2009.01.059 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, TSLP production by human BSM cells stimulated with TNF (25 ng/mL), IL-1α (10 ng/mL), IL-4, and IL-13 (50 ng/mL each). Data are representative of 4 experiments. B, Time course of TSLP production by BSM cells stimulated with IL-1/TNF. Data are representative of 2 experiments. C, Supernatants of IgE/anti-IgE–activated primary MCs (50% vol/vol), prepared as in Allakhverdi et al,5 with or without antibody to TNF or isotype control (10 μg/mL each). Data are representative of 4 experiments. ∗P < .05, ∗∗∗P < .001. Unstim, Unstimulated; SNT, supernatant. Journal of Allergy and Clinical Immunology 2009 123, 958-960.e2DOI: (10.1016/j.jaci.2009.01.059) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 BSM cells activate human MCs. BSM cells were activated with TNF (25 ng/mL) and IL-1α (10 ng/mL), and their supernatants (50% vol/vol) were used to stimulate MCs. The production of IL-5 and IL-13 were measured after 16 hours of culture in the presence or absence of anti-TSLP or isotype control mAbs (each at 10 μg/mL). Data represent the mean ± SEM of 13 experiments. ∗∗P < .01. Unstim, Unstimulated; SNT, supernatant. Journal of Allergy and Clinical Immunology 2009 123, 958-960.e2DOI: (10.1016/j.jaci.2009.01.059) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

TSLP production by human BSM cells stimulated with ligands of Toll-like receptors (TLRs) LPS (1 μg/mL), peptidoglycan, poly I:C, and flagellin (each at 100 μg/mL; A); the common allergens house dust mite (HDM), intact grains of ragweed (25 μg/mL), extract of molds (1:100 dilution), and the neuropeptides brain-derived nerve factor (BDNF), nerve growth factor β (NGF-β), and neurotrophin 3 (NT3; each at 50 ng/mL; B); and the MC mediators tryptase (0.5 ng/mL), histamine, leukotriene C4(LTC4), prostaglandin D2(PGD2), and prostaglandin E2 (PGE2; each at 1 μg/mL; C). Data are representative of 2 experiments. Unstim, Unstimulated. Journal of Allergy and Clinical Immunology 2009 123, 958-960.e2DOI: (10.1016/j.jaci.2009.01.059) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

The carryover of IL-1 in the culture supernatant of IL-1/TNF–stimulated BSM cells is required for the response of MCs to TSLP. BSM cells were activated with TNF (25 ng/mL) and IL-1α (10 ng/mL), and their supernatants (50% vol/vol) were used to stimulate MCs. The production of IL-5 was measured after 16 hours of culture in the presence or absence of anti-IL-1α, anti-TNF, and their respective isotype control mAbs (each at 10 μg/mL). MCs activated with IL-1/TNF and exogenous TSLP (10 ng/mL) were used as a positive control. Data represent the mean ± SD of triplicates and are representative of 2 experiments. Journal of Allergy and Clinical Immunology 2009 123, 958-960.e2DOI: (10.1016/j.jaci.2009.01.059) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions