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Abstract
The in vitro effect of pharmacologic concentrattons (10−8 to 10−6 mol/L) of veraparnil on human neutrophil migration and response to chemotactic signals
was examined. Human neutrophile were preinculated (15 minutes) in verapamil and then
assayed for chemotactic response to formyl-methionyl-leucyl-phenylalanine (f-Met-Leu-Phe)
(10−8 mol/L). Cell viability was not affected by verapamil treatment. Verapamil-treated
cells displayed 40% to 50% reductions in directed migration at all concentrations
(P < 0.02). Activated random migration. (chemokinesis) was also impaired by verapamil
treatment, but random locomotion was not affected except at a high concentration (10−6 mol/L). This pharmacologic action of verapamil was not rapidly revenlbie by washing
cells free of drug, but It was necessary that cells be exposed to drug before the
chemotactic signal. In addition to f-Met-Leu-Phe, chemotactic response to activated
human serum was also reduced for neutrophils. Several experiments were antagonist.
Calcium antagonist binding-site assays using radiolbeled dihydropyridlnes provided
no evidence for the presence of calcium channels in neutrophil membranes. Also, 45Ca2+ uptake assays demonstrated increased uptake of 45Ca2+ by f-Met-Leu-Phe-stimulated neutrophils, but no effect on uptake by verapamil exposures
(10−6 mol/L). Finally, the cytosalic calcium chelating dye, quin 2 acetomethoxy ester (quin
2), was used as a fluorscent indicator to measure cytosolic Ca2+ concentrations, [Ca2+], in neutrophils. Verapamil exposures over a wide concentration range (10−6 to 10−4 mol/L) did not affect resting [Ca2+] or [Ca2+] transients after f-Met-Leu-Phe (10−8 mol/L) stimulation. We conclude that human neutrophils exhibited significant decreases
in chemotactic response after exposure to concentrations of verapamil relevant to
serum concentrations achieved in the clinical setting. Furthermore, we suspect that
this pharmacologic effect is mediated by mechanisms other than calcium antagonism.
Abbreviations:
BSA (bovine serum albumin), 45Ca2+ (calcium 45), DMSO (dimethyl sulfoxide), EGTA (ethyleneglycol bis-(β-aminoethyl ether-N,N,N′,N′-tetraacetic acid), f-Met-Leu-Phe (formyl-methionyl-leucyl-phenylalanine), GBSS (Gey's balanced salt solulion), HEPES (N-2-hydroxyethylpi-perazine-N′-2-ethanesulfonic acid), quin 2-AM (quin 2 acetomethoxy ester)To read this article in full you will need to make a payment
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References
- Mechanism of action of calcium-channel-blocking agents.N Engl J Med. 1982; 307: 1618-1627
- Comparative pharmacology of calcium antagonists: nifedipine, verapamil and diltiazem.Am J Cardiol. 1980; 46: 1047-1058
- Calcium blockers and bronchoconstriction.Am Rev Respir Dis. 1983; 127: 673-674
- Voltage-dependent calcium channels in glial cells.Science. 1984; 226: 1345-1347
- Voltage-gated Ca2+ channel in mouse myeloma cells.in: Proc Natl Acad Sci USA. 80. 1983: 2240-2242
- The roles of intracellular and extracellular calcium in glucose-stimulated biphasic insulin release by rat islets.J Clin Invest. 1978; 62: 451-458
- Modification of allergic bronchoconstriction by a calcium antagonist: mode of action.Am Rev Respir Dis. 1983; 127: 675-679
- The effect of Ca2+ and Mg2+ on the chemotactic responsiveness and spontaneous motility of rabbit polymorphonuclear leukocytes.Z Immunitaetsforsch. 1972; 143: 466-476
- The regulatory role of divalent cations in human granulocyte chemotaxis: evidence for an association between calcium exchanges and microtubule assembly.J Cell Biol. 1974; 62: 594-609
- Calcium influx requirement for human neutrophil chemotaxis: inhibition by lanthanum chloride.Science. 1976; 193: 905-907
- Myosin in polymorphonuclear leukocytes.J Biol Chem. 1973; 248: 8288-8294
- Control of cytoplasmic actin gel-sol transformation by gelsolin, a calcium-dependent regulatory protein.Nature. 1979; 281: 583-586
- Role of extracellular calcium and neutrophil degranulation responses to 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine.Am J Pathol. 1981; 105: 107-113
- Interference of the calcium antagonists verapamil and nifedipine with lysosomal enzyme release from rabbit polymorphonuclear leukocytes.Drug Res. 1982; 32: 1417-1420
- Generation of superoxide radicals by human peripheral neutrophils activated by chemotactic factor. Evidence for the role of calcium.J lab clin med. 1979; 93: 583-593
- Influence of drugs that block calcium channels on the microbicidal function of human neutrophils.J Infect Dis. 1985; 151: 15-22
- Cytochalasin B facilitates the inhibition of human polymorphonuclear leukocyte generation of superoxide by verapamil.J lab clin med. 1984; 103: 949-958
- Differences in polymorphonuclear leukocyte aggregating responses among several species in response to chemotactic stimulation.J lab clin med. 1980; 96: 213-221
- The effect of verapamil and other calcium antagonists on chemotaxis of polymorphonuclear leukocytes.Biochem Pharmacol. 1984; 33: 35-39
- Clinical relevance of verapamil plasma levels in stable angina pectoris.Am J Cardiol. 1982; 50: 1180-1184
- Isolation of mononuclear cells and granulocytes from human blood.Scand J Clin Lab Invest. 1968; 21 ([Suppl]): 77-89
- Influence of immunosuppression on alveolar macrophage chemotactic activities in guinea pigs.Am Rev Respir Dis. 1981; 123: 299-304
- Relationship of binding of a calcium channel blocker to inhibition of contraction in intact cultured embryonic chick ventricular cells.Circ Res. 1983; 53: 539-543
- The oligopeptide chemotactic factor receptor on human polymorphonuclear leukocyte membranes exists in two affinity states.Biochem Biophys Res Commun. 1982; 106: 442-449
- Protein measurement with the Folin phenol reagent.J Biol Chem. 1951; 193: 265-275
- Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes.in: Proc Natl Acad Sci USA. 74. 1977: 1204-1208
- Transport of sodium, potassium, and calcium across rabbit PMN membranes, effect of chemotactic factors.J Cell Biol. 1977; 73: 428-444
- Calcium homeostasis in intact lymphocytes. Cytoplasmic free calcium monitored with a new intracellularly trapped fluorescent indicator.J Cell Biol. 1982; 94: 325-334
- Sites of action of Ca channel inhibitors.Biochem Pharmacol. 1983; 32: 3499-3507
- Stimulus response coupling in the human neutrophil. I. Kinetic analysis of changes in calcium permeability.J Biol Chem. 1984; 259: 4070-4075
- Abnormal phagocyte chemotaxis: pathophysiology, clinical manifestations, and management of patients.Rev Infect Dis. 1981; 3: 1196-1220
- Role of calcium and phosphoinositides in the actions of certain hormones and neurotransmitters.J Clin Invest. 1985; 75: 1753-1757
- Release of inflammatory mediators from stimulated neutrophils.N Engl J Med. 1980; 303: 27-34
- Calcium channel modulation by neurotransmitters. enzymes and drugs.Nature. 1983; 301: 569-574
- Molecular pharmacology of the calcium channel: evidence for subtypes, multiple drug receptor sites, channel subunits, and the development of a radioiodinated 1,4-dihydropyridine calcium channel label, 125I Iodipine.J Cardiovasc Pharmacol. 1984; 6: S608-S619
- Voltage-gated potassium channels are required for human T lymphocyte activation.J Exp Med. 1984; 160: 369-385
- Effect of the calcium antagonist verapamil on human leucocyte sodium transport in vitro.Clin Sci. 1985; 68: 239-241
- The role of protein kinase C in cell surface signal transduction and tumour promotion.Nature. 1984; 308: 694-698
Article info
Publication history
Accepted:
April 1,
1986
Received:
October 22,
1985
Footnotes
☆Supported in part by Public Health Service Grants HL 21997 and HL 00691.
Identification
Copyright
© 1986 Published by Elsevier Inc.
