Genome-wide expression profiling of B lymphocytes reveals IL4R increase in allergic asthma Marien Pascual, PhD, Sergio Roa, PhD, Asunción García-Sánchez,

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Genome-wide expression profiling of B lymphocytes reveals IL4R increase in allergic asthma Marien Pascual, PhD, Sergio Roa, PhD, Asunción García-Sánchez, PhD, Catalina Sanz, PhD, Laura Hernandez-Hernandez, PhD, John M. Greally, MD, PhD, Félix Lorente, MD, PhD, Ignacio Dávila, MD, PhD, María Isidoro-García, MD, PhD Journal of Allergy and Clinical Immunology Volume 134, Issue 4, Pages 972-975 (October 2014) DOI: 10.1016/j.jaci.2014.05.015 Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, DESeq analysis of RNA-seq data revealed a number of Ensembl transcripts to be differentially expressed between allergic and nonallergic groups. A preliminary low-stringency filter for P < .05 was used to highlight significant transcripts in red. The logarithm (base 2) of the fold change from control to allergic patients is represented by the y-axis. Normalized abundance of the transcripts (overall base mean values, ie, overall mean of read counts normalized to the library size of each sample) is shown on the x-axis. B, Heatmap showing the Euclidean distances between the samples to illustrate hierarchical clustering by using the distance matrix of the top 50 differentially expressed transcripts (P < .025). C, Significant IPA Top Diseases and BioFunctions associated with the top 50 transcripts. D, Fold increase (mean ± SEM) in the expression of candidate genes in allergic CD19+ cells (left y-axis) as measured by RNA-seq or qRT-PCR. Abundance of transcripts (base mean reads) in allergic CD19+ cells as measured by RNA-seq (right y-axis). E and F, Expression of IL4R transcripts relative to GAPDH and a standard calibrator by qRT-PCR in purified CD19+ cells (Fig 1, E) or in bulk PBMCs (Fig 1, F). Each dot represents a sample from a human subject. G, Description of the control and allergic groups of subjects included in this study. H, Linear regression of serum IgE levels and IL4R expression in the peripheral blood of 26 subjects. *The slope significantly deviates from zero (P = .0342) when constrained across the median of controls. I, Genotype frequencies of 576Q>RIL4R single-nucleotide variant (SNV: CAG>CGG), as explored by PCR amplification followed by Sanger sequencing, and their correlation with IL4R expression by qRT-PCR in PBMCs or CD19+ cells. J, Histograms of methylation data (reanalysis from Pascual et al4) in the approximately 70-kb region comprising the IL4R human locus and visualized using the UCSC Genome Browser. Deflections below the zero line represent increased methylation, whereas deflections above the line represent relative hypomethylation. A single CpG island is highlighted at the promoter region. GAPDH, Glyceraldehyde 3-phosphate dehydrogenase. Journal of Allergy and Clinical Immunology 2014 134, 972-975DOI: (10.1016/j.jaci.2014.05.015) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions