Inhibition of cPLA(2) translocation and leukotriene C-4 secretion by fluticasone propionate in exogenously activated human eosinophils

We examined the effect of the highly lipophilic corticosteroid, fluticasone propionate (FP), in causing (1) inhibition of nuclear translocation of cyt osolic phospholipase A(2) (cPLA(2)), and (2) blockade of leukotriene C-4 (L TC4) synthesis in isolated human eosinophils in vitro. Eosinophils were iso lated from peripheral blood, treated with either buffer or 10(-10) M to 10( -6) M FP in the presence of 10 pg/ml human recombinant interleukin-5 (rhIL- 5) and activated with formyl-met-leu-phe (FMLP) + cytochalasin B (CB). At 2 4 h, stimulated LTC4 secretion from eosinophils was unchanged; however, whe n corrected for cell viability, LTC4 secretion decreased from 1,429 +/- 327 pg/10(6) cells to 762 +/- 113 pg/10(6) cells for eosinophils treated for 4 8 h with greater than or equal to 10(-8) M FP (p < 0.003). FMLP/CB-stimulat ed translocation of cPLA(2) to the nuclear envelope assessed by specific im munohistochemical staining also was blocked by FP. By contrast, membrane ex pression of annexin-1, which was not minimal at 30 min, was substantial at 48 h for eosinophils treated with > 10(-10) M FP, and inhibition of LTC4 sy nthesis was reversed by exogenous arachidonic acid (AA). We find that FP ca uses a decrease in stimulated eosinophil secretion of LTC4 that is regulate d by phospholipase A(2) (PLA(2)). Inhibition of LTC4 synthesis precedes the global cytotoxic effects of FP as indicated by the simultaneous upregulati on of annexin-1 expression. Inhibited stimulated secretion corresponds to i nhibited translocation of cPLA(2) to the nuclear envelope during cellular a ctivation.