In vitro differentiated Th2 cells
Th2 cells were differentiated from peripheral blood
CD4+ T cells, as we previously described
[35].
CRTh2+CD4+ T cells (Th2 cells) were
maintained (2 x 106 cells/mL) on cycles of IL-2 (5-10
ng/mL) and plate bound antibody against CD3 and CD28 (3 days, 1µg/ml)
followed by IL-2 alone (4 days). For experiments with glucocorticoid and
estrogen, primary Th2 cells (1.3 x 106 cells/mL) were
treated (24 hours) with dexamethasone (DEX), the ERα agonist propyl
pyrazole triol (PPT) or β-estradiol (E2). To assess Th2 cell response to
CRTh2 activation, Th2 cells were pre-treated with DEX and/or PPT (24
hours), washed and re-plated with PGD2 (24 hours). Thesein vitro experiments were approved by the Ethics Review Boards at
the University of Alberta (00000942) and Western University (106770).