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Season of birth shapes neonatal immune function

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1 Season of birth shapes neonatal immune function
Anna Hammerich Thysen, PhD, Morten Arendt Rasmussen, PhD, Eskil Kreiner-Møller, PhD, Jeppe Madura Larsen, PhD, Nilofar Vahman Følsgaard, PhD, Klaus Bønnelykke, PhD, Jakob Stokholm, PhD, Hans Bisgaard, MD, DMSc, Susanne Brix, PhD  Journal of Allergy and Clinical Immunology  Volume 137, Issue 4, Pages e13 (April 2016) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Individual cord blood immune cells in relation to time of birth. Fourier series models visualize the seasonal pattern of circulating immune cell subsets in relation to birthdate. Individual immune cell counts are expressed as z scores to support visual evaluation of the seasonal variation between cell types. Minima and maxima of the Fourier curves are indicated. F, Fall; S, spring; Su, summer; W, winter. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Cord blood immune cell profiles in association with birth season. A partial least squares discrimination model representing cord blood immune cell counts in relation to season of birth is shown. A, In the score plot each dot represents the composition of immune cell subsets in a single infant. Shaded areas show the mean ± 1 SD of immune subsets of all infants born in the summer (red), fall (yellow), or winter (blue). B, The loading plot depicts the pattern of cord blood immune cells separating infants by birth season. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Cytokine and chemokine levels in upper airway lining fluid in relation to birthdate. Fourier series functions visualize the seasonal pattern of airway inflammatory mediators in relation to birthdate. Individual immune mediators are expressed as z scores to enable comparison of seasonal variation between mediators. Minima and maxima of the Fourier curves are indicated. F, Fall; S, spring; Su, summer; W, winter. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Upper airway inflammatory mediator profiles in association with birth season. A partial least squares discrimination model separating cytokine and chemokine profiles in the airway mucosal lining fluid by birth season is shown. A, In the score plots each dot represents the composition of airway immune mediators in 1 infant, and shaded areas show the mean ± 1 SD of immune subsets of all infants born in the spring (red), summer (green), fall (blue), and winter (purple). B, The loading plot depicts the pattern of airway immune mediators separating infants by birth season. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Correlation between cord blood immune cells and upper airway immune mediators. The correlation plot shows Spearman rho correlation coefficients between cord blood immune cells (left) and upper airway mucosal immune mediators at 1 month of age (top). The correlation coefficients range from −0.3 (dark blue) to 0.3 (dark red). Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Neonatal immunity in relation to birth season. Schematic presentation of the overall discrimination of circulating immune cells and airway inflammatory mediators in relation to birth seasons. Type 1 (red), type 2 (blue), and type 17 (purple) immune polarities, as well as recently activated immune mediators (orange), are encircled. Act., Activated; Eos., eosinophils; Neu., neutrophils. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E1 Gating strategy. Cord blood immune cell phenotypes were labeled in 5 separate tubes. Each tube is initially gated on forward scatter (FSC)–(SSC) properties, and singlets are selected based on area versus height parameters of forward scatter. Flow-Count beads, granulocyte subsets, and B cells are selected in tube 1. γδ, αβ, CD4, and CD8 T cells and Treg cells are quantified in tube 2, and iNKT cells are quantified in tube 3. Subsets of DCs and monocytes are gated in tube 4, whereas NK cells are detected in tube 5. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E2 Flowchart for the study population.
Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E3 Upper airway inflammatory mediator profiles in association with birth season. LV1 versus LV2 of a PLSDA model separating airway immune mediators according to birth season is shown. Each dot represents 1 child and is colored according to birth season (A), and the corresponding pattern of airway immune mediators is depicted in B. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E4 Subanalysis of upper airway inflammatory mediators and birth season in which the data are stratified based on the presence or absence of concurrent bacteria/viruses. LV2 versus LV3 of a PLSDA model separating airway immune mediators according to birth season in children with no concurrent bacterial or viral colonization is shown. The same model for children with concurrent presence of bacteria or viruses is then rotated onto the noncolonized model. Interconnected lines show how each airway inflammatory mediator shifts from the noncolonized (n = 375, squares) to the colonized (n = 195, stars) model. Journal of Allergy and Clinical Immunology  , e13DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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