Eosinophil production of prostaglandin D2 in patients with aspirin-exacerbated respiratory disease

Background: Aspirin-exacerbated respiratory disease (AERD) differs from aspirin-tolerant disease in part because of eosinophilic tissue infiltration and overexpression of arachidonic acid metabolic pathway components that lead to enhanced secretion of cysteinyl leukotrienes and prostaglandin (PG) D-2 observed constitutively and paradoxically in response to aspirin and other COX inhibitors. We have previously demonstrated the capacity of IFN-gamma to drive cysteinyl leukotriene expression and response. Objective: We investigated eosinophils as a source of PGD(2) production in patients with AERD. Methods: Eosinophils were enriched from tissue and peripheral blood obtained from control subjects, patients with aspirin-tolerant disease, and patients with AERD. mRNA was extracted and evaluated for expression of hematopoietic prostaglandin D synthase (hPGDS). Expression of hPGDS protein was confirmed with Western hybridization and immunofluorescence staining. Cells were stimulated with aspirin, and secretion of PGD(2) was quantified. CD34(+) progenitor cells were isolated and matured into eosinophils in the presence or absence of IFN-gamma and hPGDS mRNA, and PGD(2) release was measured. Results: Gene expression analysis revealed that eosinophils from tissue and blood of patients with AERD display increased levels of hPGDS compared with asthmatic and control samples. Western hybridization confirmed the increase in hPGDS mRNA translated to increased protein expression. Immunofluorescence confirmed mast cells and eosinophils from tissue of patients with AERD and asthma demonstrated hPGDS expression, with higher levels in eosinophils from patients with AERD. Incubation of eosinophils from blood and tissue with aspirin stimulated PGD(2) release. IFN-gamma-matured eosinophil progenitors showed enhanced hPGDS expression and increased levels of PGD(2) release at baseline and after aspirin stimulation. Conclusions: In addition to mast cells, eosinophils represent an important source of PGD(2) in patients with AERD and identify a new target for therapeutic intervention.