Inhibition of interleukin-8-activated human neutrophil chemotaxis by thapsigargin in a calcium- and cyclic AMP-dependent way

Chemotactic migration of human neutrophils, induced by interleukin-8 (IL-8) or other activators, was inhibited by thapsigargin in the high nanomolar range. The degree of inhibition depended on the type of activator. Other inhibitors of Ca2+-ATPases associated with intracellular calcium stores, such as cyclopiazonic acid and 2,5-di-(tert-butyl)-1,4-benzohydroquinone, equally inhibited IL-8-activated migration. Inhibition of migration by thapsigargin and the other ATPase inhibitors occurred only in the presence of extracellular Ca2+; migration was not inhibited in the presence of EGTA. La3+ reversed thapsigargin-induced inhibition to a large degree; other calcium channel blockers gave a partial reversal (econazole, verapamil, and SK&F 96365) or had no effect (gadolinium chloride and Ni2+). Using electroporated cells and Ca buffers, it was shown that inhibition started at about 0.2 mu M and was complete at a cytosolic Ca concentration of about 2 mu M. it appears that under certain conditions the thapsigargin-induced influx of extracellular calcium, causing relatively high local calcium concentrations, initiates or permits a process which may be detrimental to chemotactic migration. Cyclic AMP (cAMP; adenosine 3',5'-cyclic monophosphate) is probably involved in this process, because thapsigargin increased the cAMP level and cAMP inhibited IL-8-activated migration in a calcium-dependent way. The hypothesis that cAMP is involved in the effect of thapsigargin on migration is supported by the finding that very low concentrations of thapsigargin stimulate neutrophil migration in the absence of other chemoattractants. The results suggest that thapsigargin causes a (compartmentalized) increase in cAMP, which results in a calcium-dependent modulation of migration. BIOCHEM PHARMACOL 59;4:369-375, 2000. (C) 2000 Elsevier Science Inc.