Integrative transcriptomic analysis in human and mouse models of anaphylaxis identifies upregulation of cell movement, migration and neuroinflammatory signalling pathway

The underlying mechanisms of IgE-mediated anaphylaxis remain poorly understood. Furthermore, it remains to be determined to what extent findings from animal/mouse models reflects the pathophysiological mechanism in human. Therefore, to better characterize the mechanisms leading to potentially lethal events, analysis of global transcriptional changes (RNA-sequencing) in peripheral blood human samples in patients with anaphylaxis presenting at the emergency department and in patients with anaphylaxis during double-blind placebo-controlled food challenges (DBPCFC) to peanut, was performed. Mouse samples with different severity of IgE-mediated food-induced anaphylaxis were also included in RNA-sequencing analyses. Overall design: Emergency Department Study (ED study). We recruited 15 patients (8 female patients; age, 20-79 years) presenting with an acute episode of anaphylaxis to the ED of University Hospital Golnik, Slovenia (Table E1). Reaction severity was graded according to the Mueller criteria7. We collected blood samples at presentation to the ED and 7-14 days and/or 1 month after the anaphylactic episode (Table E1). Peanut Allergy Study (DBPCFC study). We recruited 11 patients with peanut allergy in whom allergy was confirmed by using DBPCFCs as described in details elsewhere8. Blood samples were collected before the challenge, at the cessation of the challenge because of the onset of objective symptoms, and 2 to 5 hours after the challenge (Table E2). Mouse oral antigen-induced IgE mediated anaphylaxis study (Mouse study). IgE-mediated anaphylaxis was induced in mice as described previously7, and whole blood samples were obtained after a challenge from 23 mice with different severity: allergic/no anaphylaxis (IgE anti-TNP+saline, 7 mice), mild anaphylaxis (IgE anti-TNP+OVA-TNP, 8 mice), severe anaphylaxis (IgE anti-TNP+OVA-TNP+IL-4 complex, 8 mice). This study was conducted in accordance with the amended Declaration of Helsinki. Ethical approval was obtained from the Slovenian National Medical Ethics Committee (ED study) and the London Central Research Ethics Committee (DBPCFC study). All subjects provided written informed consent. All mice were maintained in a barrier facility, and animals were handled under approved protocols of the Institutional Animal Care and Use Committee from Cincinnati Children's Hospital Medical Center.