Abstract
We have recently described a flow cytometry technique, whose sensitivity allows direct measurements of latent times before the onset of aggregation, and of rates, maximal extents, and reversibility of aggregation (J Leuk Biol 50:434, 1991). We report here that activators which stimulate sustained cellular signaling associated with increases in intracellular calcium (ionomycin) or protein kinase C activation (phorbol myristate acetate, PMA) cause complete (> or = 98%) and irreversible neutrophil aggregation, with latent times for the onset of aggregation inversely proportional to the activator concentration. In contrast, the receptor-specific activators leukotriene B4 (LTB4), formyl peptide FMLP, and platelet-activating factor (PAF) gave only partial and reversible aggregatory responses, limited by the following similar properties: latent times of 4.5 seconds +/- 1.5 seconds, independent of activator concentration; similar concentrations for onset of aggregation (approximately 1 nmol/L) that increased over a similar broad range of activator concentration, with one-half maximal rates of aggregation at 10 nmol/L to 30 nmol/L, corresponding to reported dissociation constant values; comparable limited recruitment and spontaneous reversibility of aggregation; absence of interactivator synergism; and similar exponential decays in activated cell stickiness (refractoriness), with t1/2 = 15 to 30 seconds. Variable cross- desensitization was seen between LTB4 and FMLP depending on donor and activator concentrations. In vivo, these properties are expected to provide localization of the aggregatory response, minimizing the otherwise detrimental effects of circulating activated neutrophils.
Randomized trial of a selective inhibitor of thromboxane A2 synthetase, (E)-7-phenyl-7-(3-pyridyl)-6-heptenoic acid (CV-4151), for prevention of restenosis after coronary angioplasty.