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Figure 5


Fig. 5. Antigens, anti-Fc{epsilon}Rs and anti-IgE increase cytosolic Ca2+ levels and CaN activity in human neutrophils. (A) Fura-2-AM-loaded neutrophils from an allergic patient sensitised to G3 were treated at the times indicated with 10 µg/ml anti-IgE ({alpha}-IgE) (1), 10 µg/ml G3 (2), 50 nM fMLP (3), 5 µg/ml anti-galectin-3 ({alpha}-galectin-3) (4), 5 µg/ml anti-Fc{epsilon}RI ({alpha}-Fc{epsilon}RI) (5), or 5 µg/ml anti-Fc{epsilon}RII ({alpha}-Fc{epsilon}RII) (6), and cytosolic Ca2+ was spectrofluorometrically recorded for 2 minutes. Each graph is representative of a set of three experiments that yielded similar results. (B) Neutrophils from an allergic patient sensitised to E1 were incubated with 10 µg/ml anti-IgE ({alpha}-IgE), 10 µg/ml E1 antigen, or unspecific goat IgG (IgG) for the indicated times. Then, the cells were lysed, and CaN activity was assayed. Values are the means ± s.e.m. from four separate experiments performed in triplicate.





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