Immune polarization of type 1 and type 2 human eosinophils

Eosinophils have specific immune phenotypes in type 2 and type 1 environments both in vitro and in translational models of disease in vivo. RNA sequencing was completed on human blood eosinophils cultured with cytokines that represent a Th2 (IL-33, IL-4, GM-CSF) or Th1 (TNFa/IFN?)-associated environment cytokines. Overall design: Eosinophils were isolated from the blood of otherwise healthy human donors (no allergy, atopy, pregnancy, 0r autoimmunity) and cultured for 18 hrs in different conditions prior to RNA isolation for RNA sequencing. Eosinophils were cultured at 2 million cells per ml in standard RPMI-1640+10% FBS + 1X streptomycin-penacllin-glutamine and split into three conditions: E0: [IL-5 (1ng/mL)], E1: [IL-5 (1ng/mL)+ TNF-alpha (15ng/mL) + IFN-gamma (15ng/,L); E2: [ Il-5 (1ng/mL) +IL-4 (10ng/mL )+ GM-CSF (10ng/mL) + IL-33 (50ng/mL) for 18 hrs and then isolated for RNA and RNA sequencing.