Histamine H1-receptors in HL-60 monocytes are coupled to Gi-proteins and pertussis toxin-insensitive G-proteins and mediate activation of Ca2+ influx without concomitant Ca2+ mobilization from intracellular stores

  • Roland Seifert
  • Lore Grünbaum
  • Günter Schultz


The results of binding studies suggest the presence of histamine H1-receptors in human monocytes, but it is not known whether these receptors are functionally active. This prompted us to study the effects of histamine (HA) on cytosolic Ca2+ concentration ([Ca2+]i) and superoxide anion (O2 ) formation in HL-60 cells differentiated towards monocytes with 1α,25-dihydroxychole-calciferol. In HL-60 monocytes, HA increased [Ca2+]i with a half-maximal effect at 8 μM and a maximum at 30–100 μM. Pertussis toxin (PTX) partially inhibited the stimulatory effects of HA on [Ca2+]i. Betahistine, a weak partial H1-receptor agonist, also increased [Ca2+]i, whereas H2- and H3-receptor agonists were ineffective. H1- but not H2- and H3-receptor antagonists inhibited HA induced rises in [Ca2+]i. HA-induced rises in [Ca2+]i were desensitized in a homologous manner and were also inhibited by the activator of protein kinase C, 4\-phorbol 12-myristate 13-acetate. Various protein kinase C inhibitors did not interfere with homologous desensitization. The stimulatory effects of HA on [Ca2+]i were completely dependent on the presence of extracellular Ca2+ and were inhibited by the blocker of non-selective cation (NSC) channels, 1-{\-[3-(4-methoxyphenyl)propoxyl]-4-methoxyphenethyl}-1 H-imidazole hydrochloride (SK & F 96365). HA was much less effective than the chemotactic peptide, N-formyl-l-methionyl-l-leucyl-l-phenylalanine (fMLP), to induce rises in [Ca2+]i. Unlike fMLP, HA did not activate O2 formation. Our data indicate that HL-60 monocytes possess H1-receptors coupled to heterotrimeric regulatory guanine nucleotide-binding proteins (G-proteins) of the Gi-family and PTX-insensitive G-proteins which mediate activation of NSC channels without concomitant activation of Ca2+ mobilization from intracellular stores, that homologous desensitization of HA-induced Ca2+ influx is independent of protein kinase C and that the stimulatory effect of HA on Ca2+ influx is too small to result in activation of O2 formation.

Key words

HL-60 monocytes Histamine H1-receptors G-proteins Pertussis toxin Non-selective cation channels Superoxide anion formation 


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  1. Aloj SM, Liguoro D, Klang JG, Smallridge RC (1993) Purinergic (P2) receptor-operated calcium entry into rat thyroid cells. Biochem Biophys Res Commun 195:1–7PubMedCrossRefGoogle Scholar
  2. Alonso Torre SR, Alvarez J, Montero M, Sanchez A, Garcia-Sancho J (1993) Control of Ca2+ entry into HL60 and U937 human leukemia cells by the filling state of the intracellular Ca2+ stores. Biochem J 289:761–766PubMedGoogle Scholar
  3. Amatruda TT, Steele DA, Slepak VZ, Simon MI (1991) Gα16, a G protein α subunit specifically expressed in hemopoietic cells. Proc Natl Acad Sci USA 88:5587–5591PubMedCrossRefGoogle Scholar
  4. Burde R, Seifert R, Buschauer A, Schultz G (1989) Histamine inhibits activation of human neutrophils and HL-60 leukemic cells via H2-receptors. Naunyn Schmiedebergs Arch Pharmacol 340: 671–678PubMedCrossRefGoogle Scholar
  5. Burde R, Buschauer A, Seifert R (1990) Characterization of histamine H2-receptors in human neutrophils with a series of guanidine analogues of impromidine. Are cell type-specific H2-receptors involved in the regulation of NADPH oxidase? Naunyn Schmiedebergs Arch Pharmacol 341:455–461PubMedCrossRefGoogle Scholar
  6. Buschauer A (1989) Synthesis and in vitro pharmacology of arpromidine and related phenyl(pyridylakyl)guanidines, a potential new class of positive inotropic drugs. J Med Chem 32:1963–1970PubMedCrossRefGoogle Scholar
  7. Buschauer A, Schunack W, Arrang J-M, Garbarg M, Schwartz J-C, Young JM (1989) Histamine receptors. In: Williams M, Glennon RA, Timmermans PBMWM (eds) Clinical pharmacology, Vol 13. Receptor pharmacology and function. Dekker, New York, pp 293–348Google Scholar
  8. Cameron W, Doyle K, Rocklin RE (1986) Histamine type I (H1) receptor radioligand binding studies on normal T cell subsets, B cells and monocytes. J Immunol 136:2116–2120PubMedGoogle Scholar
  9. Clapham DE (1993) A mysterious new influx factor? Nature 364:763–764PubMedCrossRefGoogle Scholar
  10. Dawra R, Saluja AK, Rünzi M, Steer ML (1993) Inositol trisphosphate-independent agonist-stimulated calcium influx in rat pancreatic acinar cells. J Biol Chem 268:20237–20242PubMedGoogle Scholar
  11. Demaurex N, Lew DP, Krause K-H (1992) Cyclopiazonic acid depletes intracellular Ca2+ stores and activates an influx pathway for divalent cations in HL60 cells. J Biol Chem 267:2318–2324PubMedGoogle Scholar
  12. Didsbury JR, Uhing RJ, Tomhave E, Gerard C, Gerard N, Snyderman R (1991) Receptor class desensitization of leukocyte chemoattractant receptors. Proc Natl Acad Sci USA 88:11564–11568PubMedCrossRefGoogle Scholar
  13. Driver AG, Kukoly CA, Bennett TE (1989) Expression of histamine H1 receptors on cultured histiocytic lymphoma cells. Biochem Pharmacol 38:3083–3091PubMedCrossRefGoogle Scholar
  14. Fujimoto K, Horio Y, Sugama K, Ito S, Liu YQ, Fukui H (1993) Genomic cloning of the rat histamine H1 receptor. Biochem Biophys Res Commun 190:294–301PubMedCrossRefGoogle Scholar
  15. Gespach C, Abita J-P (1982) Human polymorphonuclear neutrophils. Pharmacological characterization of histamine receptors mediating the elevation of cyclic AMP. Mol Pharmacol 21:78–85PubMedGoogle Scholar
  16. Gespach C, Saal F, Cost H, Abita J-P (1982) Identification and characterization of surface receptors for histamine in the human promyelocytic leukemia cell line HL60. Comparison with human peripheral neutrophils. Mol Pharmacol 22:547–553PubMedGoogle Scholar
  17. Gespach C, Cost H, Abita J-P (1985) Histamine H2 receptor activity during the differentiation of the human monocytec-like cell line U-937. Comparison with prostaglandins and isoproterenol. FEBS Lett 184:207–213PubMedCrossRefGoogle Scholar
  18. Hill SJ (1990) Distribution, properties, and functional characteristics of three classes of histamine receptors. Pharmacol Rev 42:45–83PubMedGoogle Scholar
  19. Horio Y, Mori Y, Higuchi I, Fujimoto K, Ito S, Fukui H (1993) Molecular coloning of the guinea-pig histamine H1 receptor gene. J Biochem 114:408–414PubMedGoogle Scholar
  20. Hruska KA, Bar-Shavit Z, Malone JD, Teitelbaum S (1988) Ca2+ priming during vitamin D-induced monocytec differentiation of a human leukemia cell line. J Biol Chem 263:16039–16044PubMedGoogle Scholar
  21. Jungblut PR, Seifert R (1990) Analysis by high-resolution two-dimensional electrophoresis of differentiation-dependent alterations in cytosolic protein pattern of HL60 leukemic cells. J Biochem Biophys Meth 21:47–58PubMedCrossRefGoogle Scholar
  22. Krautwurst D, Seifert R, Hescheler J, Schultz G (1992) Formyl peptides and ATP stimualte Ca2+ and Na+ inward currents through non-selective cation channels via G-proteins in dibutyryl cyclic AMP-differentiated HL-60 cells. Involvement of Ca2+ and Na+ in the activation of \-glucuronidase release and superoxide production. Biochem J 288:1025–1035PubMedGoogle Scholar
  23. Lefkowitz RJ (1993) G protein-coupled receptor kinases. Cell 74:409–412PubMedCrossRefGoogle Scholar
  24. Martiny-Baron G, Kazanietz MG, Mischak H, Blumberg PM, Kochs G, Hug H, Marmé D, Schächtele C (1993) Selective inhibition of protein kinase C isozymes by the indolcarbazole Gö 6976. J Biol Chem 268:9194–9197PubMedGoogle Scholar
  25. Merritt JE, Armstrong WP, Benham CD, Hallam TJ, Jacob R, Jaxa-Chamiec A, Leigh BK, McCarthy SA, Moores KE, Rink TJ (1990) SK&F 96365, a novel inhibitor of receptor-mediated calcium entry. Biochem J 271:515–522PubMedGoogle Scholar
  26. Monk PN, Partridge LJ (1993) Characterization of a complement-fragment-C5a-stimulated calcium-influx mechanism in U937 monocytec cells. Biochem J 295:679–684PubMedGoogle Scholar
  27. Murphy PM, Eide B, Goldsmith P, Brann M, Gierschik P, Spiegel A, Malech HL (1987) Detection of G in HL60 cells. FEBS Lett 221:81–86PubMedCrossRefGoogle Scholar
  28. Nonaka T, Mio M, Doi M, Tasaka K (1992) Histamine-induced differentiation of HL-60 cells. The role of cAMP and protein kinase A. Biochem Pharmacol 44:1115–1121PubMedCrossRefGoogle Scholar
  29. Norgauer J, Dobos G, Kownatzki E, Dahinden C, Burger R, Kupper R, Gierschik P (1993) Complement C3a stimulates Ca2+ influx in neutrophils via a pertussis-toxin-sensitive G protein. Eur J Biochem 217:289–294PubMedCrossRefGoogle Scholar
  30. Ostrem VK, Tanaka Y, Prahl J, DeLuca HF, Ikekawa N (1987) 24- and 26-homo-1,25-dihydroxyvitamin D3: Preferential activity in inducing differentiation of human leukemia cells HL-60 in vitro. Proc Natl Acad Sci USA 84:2610–2614PubMedCrossRefGoogle Scholar
  31. Pollock K, Creba J, Mitchell F, Milligan G (1990) Stimulus-response coupling in FMLP-stimulated U937 monocytes: Effect of differentiation on Gi2 expression. Biochem Biophys Acta 1051:71–77PubMedCrossRefGoogle Scholar
  32. Randriamampita C, Tsien TY (1993) Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature 364:809–814PubMedCrossRefGoogle Scholar
  33. Rosenthal W, Koesling D, Rudolph U, Kleuss C, Pallast M, Yajima M, Schultz G (1986) Identification of the 35-kDa \-subunit of guanine-nucleotide-binding proteins by an antiserum raised against transducin. Eur J Biochem 158:255–263PubMedCrossRefGoogle Scholar
  34. Rudolph U, Koesling D, Hinsch K-D, Seifert R, Bigalke M, Schultz G, Rosenthal W (1989) G-protein α-subunits in cytosolic and membranous fractions of human neutrophils. Mol Cell Endocrinol 63:143–153PubMedCrossRefGoogle Scholar
  35. Schunack W (1987) What are the differences between H2-receptor antagonists? Aliment Pharmacol Therap 1:4935–5035Google Scholar
  36. Schwaner I, Seifert R, Schultz G (1992) Receptor-mediated increases in cytosolic Ca2+ in the human erythroleukemia cell line involve pertussis toxin-sensitive and -insensitive pathways. Biochem J 281: 301–307PubMedGoogle Scholar
  37. Seligmann BE, Flechter MP, Gallin JI (1983) Histamine modulation of human neutrophil oxidative metabolism, locomotion, degranulation, and membrane potential changes. J Immunol 130:1902–1909PubMedGoogle Scholar
  38. Seifert R, Schultz G (1987) Reversible activation of NADPH oxidase in membranes of HL-60 leukemic cells. Biochem Biophys Res Commun 146:1296–1302PubMedCrossRefGoogle Scholar
  39. Seifert R, Burde R, Schultz G (1989) Activation of NADPH oxidase by purine and pyrimidine nucleotides involves G proteins and is potentiated by chemotactic peptides. Biochem J 259:813–819PubMedGoogle Scholar
  40. Seifert R, Höer A, Offermanns S, Buschauer A, Schunack W (1992) Histamine increases cytosolic Ca2+ in dibutyryl-cAMP-differentiated HL-60 cells via H1 receptors and is an incomplete secretagogue. Mol Pharmacol 42:227–234PubMedGoogle Scholar
  41. Sozzani S, Molino M, Locati M, Luini W, Cerletti C, Vecchi A, Mantovani A (1993) Receptor-activated calcium influx in human monocytes exposed to monocyte chemotactic protein-1 and related cytokines. J Immunol 150:1544–1553PubMedGoogle Scholar
  42. Strathmann MP, Simon MI (1991) Gα12 and Gα13 subunits define a fourth class of G-protean a subunits. Proc Natl Acad Sci USA 88:5582–5586PubMedCrossRefGoogle Scholar
  43. Tamaoki T, Nomoto H, Takahashi I, Kato Y, Morimoto M, Tomita F (1986) Staurosporine, a potent inhibitor of phospholipid/Ca++ dependent protein kinase. Biochem Biophys Res Commun 135: 397–402PubMedCrossRefGoogle Scholar
  44. Tohkin M, Morishima N, Iiri T, Takahashi K, Ui M, Katada T (1991) Interaction of guanine-nucleotide-binding regulatory proteins with chemotactic peptide receptors in differentiated human leukemic HL-60 cells. Eur J Biochem 195:527–533PubMedCrossRefGoogle Scholar
  45. Wenzel-Seifert K, Seifert R (1990) Nucleotide-, chemotactic peptide-and phorbol ester-induced exocytosis in HL-60 leukemic cells. Immunobiology 181:298–316PubMedCrossRefGoogle Scholar
  46. Werlen G, Belin D, Conne B, Roche E, Lew DP, Premki M (1993) Intracellular Ca2+ and the regulation of early response gene expression in HL-60 myeloid leukemia cells. J Biol Chem 268:16596–16601PubMedGoogle Scholar
  47. Yamashita M, Fukui H, Sugama K, Horio Y, Ito S, Mizuguchi H, Wada H (1991) Expression cloning of a cDNA encoding the bovine histamine H1 receptor. Proc Natl Acad Sci USA 88:11515–11519PubMedCrossRefGoogle Scholar
  48. Zingel V, Schunack W (1993) Agonisten des Histamin H1-Rezeptors: Struktur und Pharmakologie. Pharmazie 48:483–493PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Roland Seifert
    • 1
  • Lore Grünbaum
    • 1
  • Günter Schultz
    • 1
  1. 1.Institut für Pharmakologie, Universitätsklinikum Rudolf VirchowFreie Universität BerlinBerlinGermany

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