Platelet-activatingfactor-inducedchemotaxis and interleukin-5-inducedchemokinesisare mediated by different signals

Eosinophils play an important role in the pathogenesis of allergic diseases such as allergic asthma. Eosinophil migration in vitro can be divided into directed migration, or chemotaxis, and random migration, or chemokinesis. Here, we studied mtra- cellular signals involved in eosinophil migration in vitro induced by platelet-activating factor (PAF) and mterleukin-5 (IL-5), applying a Boyden chamber assay. Migration induced by PAF (10h1_1O6 M) largely consisted of chemotaxis with some chemoki- nesis, whereas IL-5 (1012_i08 M) induced chemok- inesis only. Eosinophils were depleted from intracellular and extracellular Ca2+ to study the role of Ca2' as a second messenger. Ca2+ depletion did not change PAF-mduced chemotaxis, however, IL- 5-induced chemokinesis was inhibited. Interestingly, PAF, but not IL-5, induced changes in (Ca2�J1. This rise originated mainly from internal stores. Inhibition of protein kinase A by H-89 and protein kinase C by GF 1 09203X had no effect on both forms of eosino- phil migration. Addition of the protein kinase inhibi- t 0 r 5 t au ro sp o rine signifi c antly inhibited IL-5-induced chemokinesis. Inhibition of tyrosine kinases by herbimycin A completely blocked IL-5-in- duced chemokinesis. PAF and IL-5-induced actin polymerization was studied to compare migratory responses with a migration-associated intracellular response. Ca2+ depletion significantly enhanced PAF-induced (108 M) actin polymerization, whereas IL-5-induced actin polymerization was not influ- enced. Addition of staurosporine led to an increase in F-actin. Subsequent addition of PAF or IL-5 resulted in an additive increase in F-actin content. In summary, both forms of eosinophil migration are protein kinase A and protein kinase C independent. In contrast to PAF-induced chemotaxis, IL-5-in- duced chemokinesis was found to be completely Ca2+ and tyrosine kinase dependent. J. Leukoc. Biol. 59: 347-356; 1996.

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