Original article| Volume 108, ISSUE 1, P37-43, July 1986

Verapamil impairs human neutrophil chemotaxis by a non-calcium-mediated mechanism

      This paper is only available as a PDF. To read, Please Download here.


      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.


      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

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Translational Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Braunwald E
        Mechanism of action of calcium-channel-blocking agents.
        N Engl J Med. 1982; 307: 1618-1627
        • Henry PD
        Comparative pharmacology of calcium antagonists: nifedipine, verapamil and diltiazem.
        Am J Cardiol. 1980; 46: 1047-1058
        • Fanta CH
        • Drazen JM
        Calcium blockers and bronchoconstriction.
        Am Rev Respir Dis. 1983; 127: 673-674
        • MacVicar BA
        Voltage-dependent calcium channels in glial cells.
        Science. 1984; 226: 1345-1347
        • Fukushima Y
        • Hagiwara S
        Voltage-gated Ca2+ channel in mouse myeloma cells.
        in: Proc Natl Acad Sci USA. 80. 1983: 2240-2242
        • Wolheim CB
        • Kikuchi M
        • Renold AE
        • Sharp GWG
        The roles of intracellular and extracellular calcium in glucose-stimulated biphasic insulin release by rat islets.
        J Clin Invest. 1978; 62: 451-458
        • Russi EW
        • Marchette B
        • Yerger L
        • Abraham WM
        • Ahmed T
        Modification of allergic bronchoconstriction by a calcium antagonist: mode of action.
        Am Rev Respir Dis. 1983; 127: 675-679
        • Becker EL
        • Showell HJ
        The effect of Ca2+ and Mg2+ on the chemotactic responsiveness and spontaneous motility of rabbit polymorphonuclear leukocytes.
        Z Immunitaetsforsch. 1972; 143: 466-476
        • Gallin JI
        • Rosenthal AS
        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
        • Boucek MM
        • Snyderman R
        Calcium influx requirement for human neutrophil chemotaxis: inhibition by lanthanum chloride.
        Science. 1976; 193: 905-907
        • Stossel TP
        • Pollard TD
        Myosin in polymorphonuclear leukocytes.
        J Biol Chem. 1973; 248: 8288-8294
        • Yin HL
        • Stossel TP
        Control of cytoplasmic actin gel-sol transformation by gelsolin, a calcium-dependent regulatory protein.
        Nature. 1979; 281: 583-586
        • O'Flaherty JT
        • Swendsen CL
        • Lees CJ
        • McCall CE
        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
        • Elferink JGR
        Interference of the calcium antagonists verapamil and nifedipine with lysosomal enzyme release from rabbit polymorphonuclear leukocytes.
        Drug Res. 1982; 32: 1417-1420
        • Simchowitz L
        • Spilberg I
        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
        • Kazanjian PH
        • Pennington JE
        Influence of drugs that block calcium channels on the microbicidal function of human neutrophils.
        J Infect Dis. 1985; 151: 15-22
        • Steiner RD
        • Pratt A
        • Busse WW
        Cytochalasin B facilitates the inhibition of human polymorphonuclear leukocyte generation of superoxide by verapamil.
        J lab clin med. 1984; 103: 949-958
        • Oseas RS
        • Boxer LA
        • Butterick C
        • Baehner RL
        Differences in polymorphonuclear leukocyte aggregating responses among several species in response to chemotactic stimulation.
        J lab clin med. 1980; 96: 213-221
        • Elferink JGR
        • Deierkauf M
        The effect of verapamil and other calcium antagonists on chemotaxis of polymorphonuclear leukocytes.
        Biochem Pharmacol. 1984; 33: 35-39
        • Frishman W
        • Kirsten E
        • Klein M
        • et al.
        Clinical relevance of verapamil plasma levels in stable angina pectoris.
        Am J Cardiol. 1982; 50: 1180-1184
        • Boyum A
        Isolation of mononuclear cells and granulocytes from human blood.
        Scand J Clin Lab Invest. 1968; 21 ([Suppl]): 77-89
        • Pennington JE
        • Harris EA
        Influence of immunosuppression on alveolar macrophage chemotactic activities in guinea pigs.
        Am Rev Respir Dis. 1981; 123: 299-304
        • Marsh JD
        • Loh E
        • Lachance D
        • Barry WH
        • Smith TW
        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
        • Koo C
        • Lefkowitz RJ
        • Snyderman R
        The oligopeptide chemotactic factor receptor on human polymorphonuclear leukocyte membranes exists in two affinity states.
        Biochem Biophys Res Commun. 1982; 106: 442-449
        • Lowry OH
        • Rosebrough NJ
        • Farr AL
        • Randall R
        Protein measurement with the Folin phenol reagent.
        J Biol Chem. 1951; 193: 265-275
        • Williams LT
        • Snyderman R
        • Pike MC
        • Lefkowitz RJ
        Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes.
        in: Proc Natl Acad Sci USA. 74. 1977: 1204-1208
        • Naccache PH
        • Showell HJ
        • Becker EL
        • Sha'afi RI
        Transport of sodium, potassium, and calcium across rabbit PMN membranes, effect of chemotactic factors.
        J Cell Biol. 1977; 73: 428-444
        • Tsien R
        • Pozzan T
        • Rink T
        Calcium homeostasis in intact lymphocytes. Cytoplasmic free calcium monitored with a new intracellularly trapped fluorescent indicator.
        J Cell Biol. 1982; 94: 325-334
        • Janis RA
        • Scriabine A
        Sites of action of Ca channel inhibitors.
        Biochem Pharmacol. 1983; 32: 3499-3507
        • Korchak HM
        • Rutherford LE
        • Weissmann G
        Stimulus response coupling in the human neutrophil. I. Kinetic analysis of changes in calcium permeability.
        J Biol Chem. 1984; 259: 4070-4075
        • Gallin JI
        Abnormal phagocyte chemotaxis: pathophysiology, clinical manifestations, and management of patients.
        Rev Infect Dis. 1981; 3: 1196-1220
        • Exton JH
        Role of calcium and phosphoinositides in the actions of certain hormones and neurotransmitters.
        J Clin Invest. 1985; 75: 1753-1757
        • Weissmann G
        • Smolen JE
        • Korchak HM
        Release of inflammatory mediators from stimulated neutrophils.
        N Engl J Med. 1980; 303: 27-34
        • Reuter H
        Calcium channel modulation by neurotransmitters. enzymes and drugs.
        Nature. 1983; 301: 569-574
        • Glossmann H
        • Ferry DR
        • Goll A
        • Rombusch M
        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
        • Chandy KG
        • DeCoursey TE
        • Cahalan MD
        • McLaughlin C
        • Gupta S
        Voltage-gated potassium channels are required for human T lymphocyte activation.
        J Exp Med. 1984; 160: 369-385
        • Gray HH
        • Poston L
        • Johnson VE
        • Hilton PJ
        Effect of the calcium antagonist verapamil on human leucocyte sodium transport in vitro.
        Clin Sci. 1985; 68: 239-241
        • Nishizuka Y
        The role of protein kinase C in cell surface signal transduction and tumour promotion.
        Nature. 1984; 308: 694-698