Inhibition of phospholipase A2 activity in guinea pig eosinophils by human recombinant IL-1 beta.
The effect of human rIL-1 beta on the release of arachidonic acid (AA) and on the phospholipase A2 (PLA2) activity in guinea pig eosinophils was investigated. Stimulation of [3H]AA-labeled eosinophils with the ionophore A23187 resulted in a time and concentration-dependent release of AA in parallel to hydrolysis of endogenous phosphatidylcholine (PC). Both events were abrogated by the chelation of intracellular free calcium, but not by its depletion from the medium, suggesting that the ionophore-induced AA release involves a PLA2 activity dependent on the mobilization of intracellular calcium. Addition of human rIL-1 beta (0.01 to 100 ng/ml) to eosinophils for 15 min had no effect on the release of AA induced by the ionophore. However, prolonged incubation with human rIL-1 beta (30 to 180 min) inhibited in a concentration- and time-dependent manner the release of AA and the hydrolysis of phosphatidylcholine in ionophore-stimulated eosinophils. Our results also showed that eosinophil homogenates contain a calcium-dependent PLA2 whose activity was markedly reduced when eosinophils were pretreated with human rIL-1 beta. The inhibition was time and concentration dependent and was observed in the presence of calcium and phospholipid excess. Finally, studies with Fura-2-loaded eosinophils showed that the ionophore A23187 stimulated an increase in intracellular calcium concentration that was not altered by pretreating the eosinophils with human rIL-1 beta. These results suggest that human rIL-1 beta inhibits the release of AA by eosinophils via the inhibition of a PLA2 activity and through a calcium-independent mechanism. Inhibition by human rIL-1 beta required a prolonged incubation (30 to 180 min) and was observed after its removal from the medium, suggesting that human rIL-1 beta did not interact directly with the PLA2 itself, but with a metabolic process involved with the regulation of its activity in eosinophils.
Debbaghi
A
A
Unité de Pharmacologie Cellulaire, Unité Associée Institut Pasteur, INSERM U.285, Paris, France.
Hidi
R
R
Vargaftig
B B
BB
Touqui
L
L
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D004533:Egtazic Acid / Q000494:pharmacology; D004804:Eosinophils / Q000201:enzymology*; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000494:pharmacology*; D010713:Phosphatidylcholines / Q000378:metabolism; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D054467:Phospholipases A2; D011994:Recombinant Proteins / Q000494:pharmacology
D007375:Interleukin-1; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D016718:Arachidonic Acid; D000001:Calcimycin; D004533:Egtazic Acid; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium