Maternal and cord blood miR-223 expression associates with prenatal tobacco smoke exposure and low regulatory T-cell numbers Gunda Herberth, PhD, Mario.

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Maternal and cord blood miR-223 expression associates with prenatal tobacco smoke exposure and low regulatory T-cell numbers Gunda Herberth, PhD, Mario Bauer, MD, Michaela Gasch, MSc, Denise Hinz, PhD, Stefan Röder, PhD, Sven Olek, PhD, Tibor Kohajda, PhD, Ulrike Rolle-Kampczyk, PhD, Martin von Bergen, PhD, Ulrich Sack, MD, Michael Borte, MD, Irina Lehmann, PhD Journal of Allergy and Clinical Immunology Volume 133, Issue 2, Pages 543-550.e4 (February 2014) DOI: 10.1016/j.jaci.2013.06.036 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Relationship between smoking during pregnancy and measured VOC concentrations at home (n = 617), as well as corresponding metabolites in maternal urine (n = 608). Linear regression models (shown are MRs and 95% CIs) were adjusted for renovation activities and traffic exposure during pregnancy. Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Comparison between miRNA expression in maternal blood during pregnancy (n = 322) and cord blood (n = 450, relative expression). Data are shown in box plots (first and third quartiles and medians), and the whiskers indicate ranges without outliers. P values were calculated by using the Wilcoxon test. Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Relationship between VOC concentrations in dwellings and corresponding metabolites in maternal urine and miRNA expression in blood samples during pregnancy (n = 316) and cord blood (n = 444). Linear regression models (shown are MRs and 95% CIs) were adjusted for parental history of atopy (maternal history of atopy for mothers), parental education (maternal education for mothers), and sex (only for cord blood). Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Relationship between miRNA expression in maternal and cord blood and Treg cell numbers, as detected by means of demethylation of TSDR in the FOXP3 gene in cord blood (n = 448) and pregnancy blood samples (n = 318), respectively. Linear regression models (shown are MRs and 95% CIs) were adjusted for parental history of atopy (maternal history of atopy for mothers), parental education (maternal education for mothers), maternal smoking and/or ETS exposure during pregnancy, and sex (only for cord blood). Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Prenatal environmental factors associated with alterations in Treg cell numbers at birth and risk for atopic dermatitis. Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Flow chart of the LINA study population with the analyzed subcohort. Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Immune cell–specific miRNA expression in granulocytes, stimulated and unstimulated PBMCs, Teff cells, and Treg cells was quantified by using real-time PCR. Expression levels were normalized to SNORD44 expression (fold change × 100) and referred to the maximum miRNA expression within the immune cell subsets, respectively. The highest expression of miR-223 and miR-155 was determined in granulocytes and stimulated PBMCs, respectively. Data are presented as means ± SEMs (n = 3). Journal of Allergy and Clinical Immunology 2014 133, 543-550.e4DOI: (10.1016/j.jaci.2013.06.036) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions