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Agonist-induced calcium flux, phosphoinositide metabolism, aggregation and enzyme secretion in human neutrophils

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Abstract

Formyl-methionyl-leucyl-phenylalanine (FMLP), platelet activating factor (PAF) and leukotriene B4 (LTB4) are potent activators of human neutrophils. Using human neutrophils prelabelled with the fluorescent indicator dye, Quin 2, or with [32P]-orthophosphate, we examined the effects of these stimuli on intracellular free calcium concentration, [Ca2+]i, and, on various indices of phosphoinositide metabolism, including [32P]-phosphatidic acid (PtdA) formation. The concentration-dependence of the observed changes in [Ca2+]i or [32P]-PtdA were then compared to stimulus-induced aggregation and enzyme release (β-N-acetylglucosaminidase (NAG) and lysozyme).

FMLP, PAF and LTB4 caused a concentration-dependent elevation of [Ca2+]i, aggregation and enzyme release. However, unlike FMLP and PAF, LTB4 (≦2.5 μM) did not cause significant formation of [32P]-PtdA. The concentration response curves for agonist-induced elevation of [Ca2+]i lie to the left of those for aggregation and enzyme release. FMLP and PAF also caused an elevation of [Ca2+]i at concentrations lower than those required to elicit [32P]-PtdA formation.

These observations suggest that [Ca2+]i elevationper se cannot mediate human neutrophil functional responses to FMLP, PAF and LTB4. Consequently there may exist other mediator(s) that act in concert with [Ca2+]i or are triggered by [Ca2+]i elevation to promote human neutrophil activation. Both the elevation of [Ca2+]i and the formation of these putative mediator(s) in response to LTB4 apparently occur independently of inositol phospholipid hydrolysis.

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  1. D. E. MacIntyre

    Present address: Dept. of Biochemistry & Molecular Biology, Merck Sharp & Dohme Research Laboratories, P.O. 2000, 07065, Rahway., New Jersey, USA

Authors and Affiliations

  1. Department of Pharmacology, University of Glasgow, G12 8QQ, Glasgow, Scotland, U.K.

    A. G. Rossi & D. E. MacIntyre

  2. Bioscience Department I, ICI Pharmaceuticals Division, Mereside, Alderley Park, SK10 4TG, Macclesfield, Cheshire, England, U.K.

    R. M. McMillan

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Rossi, A.G., McMillan, R.M. & MacIntyre, D.E. Agonist-induced calcium flux, phosphoinositide metabolism, aggregation and enzyme secretion in human neutrophils. Agents and Actions 24, 272–282 (1988). https://doi.org/10.1007/BF02028283

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