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Functional analysis of protective IL1RL1 variants associated with asthma risk  Vladimir Ramirez-Carrozzi, PhD, Amy Dressen, MS, Patrick Lupardus, PhD,

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Presentation on theme: "Functional analysis of protective IL1RL1 variants associated with asthma risk  Vladimir Ramirez-Carrozzi, PhD, Amy Dressen, MS, Patrick Lupardus, PhD,"— Presentation transcript:

1 Functional analysis of protective IL1RL1 variants associated with asthma risk 
Vladimir Ramirez-Carrozzi, PhD, Amy Dressen, MS, Patrick Lupardus, PhD, Brian Yaspan, PhD, Rajita Pappu, PhD  Journal of Allergy and Clinical Immunology  Volume 135, Issue 4, Pages e3 (April 2015) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 IL1RL1 protective variants confer reduced IL-33 activity. A, Schematic of IL-33 receptor complex and location of variants. IRAK, IL-1 receptor–associated kinase; MAPKK, mitogen-activated protein kinase kinase. B, Overlay of variants A433T and Q501R on crystal structure of the TLR10 TIR domain. C and D, HEK-Blue cells expressing common or protective ST2 variants were stimulated with IL-33 (Fig 1, C) or IL-1β (Fig 1, D), as indicated. EC50, Median effective concentration. E, ELISA of IL-8 secretion from purified blood eosinophils treated with IL-33. F, Quantitative RT-PCR of sST2 mRNA expression from purified blood eosinophils and basophils. G, ELISA analysis of plasma sST2 levels. Plots display means ± SEMs of 3 single clones or 4 individual donors per group. *P < .05, paired t test. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig E1 Analysis of the individual and combinatorial IL1RL1 variants on IL-33 responses. Batch clones of HEK-Blue cells expressing the common variant or permutations of the protective ST2 variants were stimulated with increasing concentrations of human recombinant IL-33 (A) or IL-1β (B) for 20 hours. Cytokine activity was measured based on induction of NF-κB/AP-1 SEAP reporter activity. Plots show means ± SDs of triplicates. Data are representative of 3 independent experiments. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E2 ST2 receptor expression analysis in HEK-Blue cells expressing IL1RL1 variants. A, Left panel, Fluorescence-activated cell sorting analysis of ST2 surface expression for each single clone cell line. Right panel, Detection of IL-1RAcP surface expression. B, Mean fluorescence intensity calculation of ST2 surface expression from the graphs shown in Fig E2, A. C, Quantitative RT-PCR measurements of IL1RL1 and IL1RAP. mRNA results are presented relative to expression of the housekeeping gene RPL19 (encoding the ribosomal protein L19). Undet., Undetectable. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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