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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 2, August, 2001 239-244

Interleukin-4 Rapidly Inhibits Calcium Transients in Response to Carbachol in Bovine Airway Smooth Muscle Cells

J. Mark Madison and Michael F. Ethier

Division of Pulmonary, Allergy and Critical Care Medicine, University of Massachusetts Medical School, Worcester, Massachusetts

To assess interleukin (IL)-4 effects on calcium signaling, bovine airway smooth-muscle (ASM) cells were loaded with fura-2 and cytosolic calcium ([Ca2+]i) was measured in single cells by digital microscopy. Human recombinant IL-4 (50 ng/ml) caused small increases in [Ca2+]i. For single cells, carbachol-stimulated calcium transients were compared before (S1) and after (S2) exposure to IL-4 or IL-13. When cells were treated with IL-4 (50 ng/ml) for 20 min, the S2/S1 ratio was 0.17 ± 0.04 (n = 7) even though IL-4 had been washed from the chamber for 10 min before the S2 response. In contrast, controls not treated with IL-4 had S2/S1 of 0.70 ± 0.04 (n = 13, P < 0.01). Lower concentrations of IL-4 variably decreased transients and IL-13 had no effect. In other experiments, 5 min of IL-4 did not immediately decrease transients but did after a 25-min delay. Goat antihuman IL-4 antibody abolished the effect of IL-4. IL-4 (50 ng/ml) also inhibited responses to caffeine (S2/S1: 0.30 ± 0.04 and 0.54 ± 0.06 for IL-4-treated versus control). We conclude that IL-4 rapidly inhibited calcium transients. Because caffeine-stimulated transients were inhibited, IL-4 may act, at least in part, by depleting calcium stores. IL-4 inhibition of cholinergic signaling may be important for modulating ASM responses during inflammation.

Abbreviations: airway smooth muscle, ASM; cytosolic calcium, [Ca2+]i; interleukin, IL; IL-4 receptor, IL-4R; inositol trisphosphate, IP3; physiologic salt solution, PSS; standard error of the mean, SEM; sarcoplasmic reticulum, SR; T helper, TH.




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