Immune polarization of type 1 and type 2 mouse 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 mouse 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 IL-5 Tg mice (NJ.1638) and cultured for 18 hrs in different conditions prior to RNA isolation for RNA sequencing. Eosinophils were cultured at 5 million cells per ml in standard RPMI-1640+10% FBS + 1X streptomycin-penacllin-glutamine. One experiment included duplicates of control eosinophils (CN) has no cytokines added conducted at the same time as eosinophils (E1) were cultured with 15ng/mL IFN-gamma and TNF-alpha. A second experiment included duplicates of IL-5 eosinophils (E0) were cultured with IL-5 10 ng/mL and eosinophils (E2) were cultured with 10ng/ml of IL-5, GM-CSF, IL-4 and 30ng/mL IL-33.