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Thymic stromal lymphopoietin does not activate human basophils

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Presentation on theme: "Thymic stromal lymphopoietin does not activate human basophils"— Presentation transcript:

1 Thymic stromal lymphopoietin does not activate human basophils
Nina Salabert-Le Guen, PhD, Caroline Hémont, PharmD, PhD, Agathe Delbove, MD, Caroline Poli, MD, PhD, Cécile Braudeau, PhD, Aurélie Fantou, PharmD, Karine Amouriaux, PhD, Gaelle Bériou, PhD, Jérôme C. Martin, PharmD, PhD, Luc Colas, MD, Vassili Soumelis, MD, PhD, Régis Josien, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 141, Issue 4, Pages e6 (April 2018) DOI: /j.jaci Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 TSLP does not activate purified blood basophils from HDs. Blood basophils and mDCs were isolated from HDs and cultured with medium, IL-3, or TSLP for expression of CD63, CD69, and CD203c on basophils (n = 4-7) (A). Cytokine and chemokine concentration in basophil supernatants (n = 4) (B). Expression of CD83 and CD86 on mDCs (n = 6) (C). Kinetics of P-STAT5 expression in basophils (top) and mDCs (bottom) (n = 3) (D). TSLPR and IL-7Rα expression on basophils (top) and mDCs (bottom) (E). MCP-1, Monocyte chemoattractant protein; MIP-1α, macrophage inflammatory protein-1α. Data are shown as mean ± SEM. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Basophils from allergic patients do not respond to TSLP and do not express IL-7Rα. Whole blood samples from HDs (empty bars, n = 5) and allergic patients (black bars, n = 8) were stimulated for 6 hours with IL-3, TSLP, anti-FcɛRI antibody, or medium and analyzed by FCM. Gating strategy on whole blood (A). Intracellular P-STAT5 expression in basophils (left) and mDCs (right) (B). CD203c (left) and CCR3 (right) expression on basophils (C). % of IL-7Rα– and TSLPR- expressing basophils (left) or mDCs (right) (D). Data are shown as mean ± SEM. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 A, CD63, CD69, and CD203c expression on freshly isolated basophils and after 24-hour stimulation with medium, IL-3, or TSLP. B, CD86 and CD83 expression on isolated mDCs after 24-hour stimulation with medium or TSLP. C, Cytokine secretion by isolated mDCs after 24-hour stimulation by medium, IL-3, or TSLP and measured by a multiplex assay. MCP-1, Monocyte chemoattractant protein; MIP-1α, macrophage inflammatory protein-1α. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 A, Histograms of P-STAT5 expression in purified basophils and mDCs after stimulation by IL-3 or TSLP. B, P-STAT5 expression in basophils after 24-hour priming followed by 15-minute stimulation with IL-3 or TSLP (n = 3-5 donors). Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Intracellular P-STAT5 expression in basophils and mDCs (A) in whole blood samples from an allergic donor, after 6-hour stimulation with IL-3, TSLP, or medium. CD203c and CCR3 expression on basophils (B) and IL-7Rα and TSLPR expression on mDCs (C) or basophils (D) in whole blood samples from an allergic donor, either fresh or after 6-hour stimulation with IL-3, TSLP, anti-FcɛRI antibody, or medium. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 A, TSLPR and IL7R mRNA expression assessed by RT-PCR in fresh and IL-3–activated basophils from allergic patients (n = 3-5 per time point). B, CD203c and CD69 expression on blood basophils isolated from allergic patients using the Stem Cell Technologies kit, either fresh or after culture with medium, IL-3, TSLP, anti-IgE mAb, or anti-FcεRI mAb for 2 hours or 22 hours (n = 3-5 per time point/condition). C, TSLPR and IL-7Rα expression on blood basophils isolated from allergic patients using the Stem Cell Technologies kit, either fresh or after culture with IL-3, TSLP, anti-IgE mAb, or anti-FcεRI mAb for 2 hours or 22 hours. Similar results were obtained in 5 different allergic individuals. A.U., Arbitray unit. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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