Tobacco use is associated with an increase in the white blood cell (WBC) count. This association has been attributed to bronchopulmonary inflammation and/or infection. It is not known if nicotine itself may play a role. The objective of this study was to determine whether nicotine itself could affect the WBC count, and to determine whether this was due to a direct effect on hematopoietic stem cells (HSC). C57Bl6J mice received nicotine orally, and measurements of the WBC count, bone marrow and spleen cellularity, and HSC count were made. To determine the functionality of HSCs, irradiated animals received bone marrow transplants from vehicle or nicotine-treated mice. Nicotine increased leukocytes in the peripheral blood, bone marrow and spleen. The peripheral red cell and platelet count were unaffected. Nicotine increased the frequency of HSC in the bone marrow. Isolated long-term HSCs from nicotine-treated mice transplanted into irradiated mice regenerated all hematopoietic cell lineages, demonstrating the functional competence of those HSCs. HSCs expressed nicotinic acetylcholine receptors (nAChRs), as documented by FITC-conjugated alpha-bungarotoxin binding. Nicotine increased soluble Kit ligand, consistent with stem cell activation. In conclusion, the data suggest a new mechanism for the increased WBC associated with tobacco use. The effect of nicotine to activate hematopoiesis may contribute to tobacco-related diseases.
Jensen EJ, Pedersen B., Frederiksen R., Dahl R.Prospective study on the effect of smoking and nicotine substitution on leucocyte blood counts and relation between blood leucocytes and lung function . Thorax1998; 53: 784-789.
2.
Corre F., Lellouch J., Schwartz D.Smoking and leucocyte-counts. Results of an epidemiological survey . Lancet1971; ii: 632-634.
3.
Sparrow D., Glynn RJ, Cohen M.The relationship of the peripheral leukocyte count and cigarette smoking to pulmonary function among adult men. Chest1984; 86: 383-386.
4.
Ensrud K., Grimm RHJ.The white blood cell count and risk for coronary heart disease. Am Heart J1992; 124: 207-213.
5.
Madjid M., Awan I., Willerson JT, Casscells SWLeukocyte count and coronary heart disease: implications for risk assessment . J Am Coll Cardiol2004; 44: 1945-1956.
6.
Haim M., Boyko V., Goldbourt U., Battler A., Behar S.Predictive value of elevated white blood cell count in patients with preexisting coronary heart disease: the Bezafibrate Infarction Prevention Study. Arch Intern Med2004; 164: 433-439.
7.
Friedman GD, Klatsky AL, Siegelaub ABThe leukocyte count as a predictor of myocardial infarction. N Engl J Med1974; 290: 1275-1278.
8.
Friedman GD, Siegelaub AB, Seltzer CC, Feldman R., Collen MFSmoking habits and the leukocyte count. Arch Environ Health1973; 26: 137-143.
9.
Pitzer JE, Del Zoppo GJ, Schmid-Schonbein GWNeutrophil activation in smokers. Biorheology1996; 33: 45-58.
10.
Koethe SM, Kuhnmuench JR, Becker CGNeutrophil priming by cigarette smoke condensate and a tobacco anti-idiotypic antibody. Am J Pathol2000; 157: 1735-1743.
11.
Calafat AM, Polzin GM, Saylor J., Richter P., Ashley DL, Watson CHDetermination of tar, nicotine, and carbon monoxide yields in the mainstream smoke of selected international cigarettes. Tob Control2004; 13: 45-51.
12.
Benowitz NL , Hukkanen J., Jacob P. 3rd.Nicotine chemistry, metabolism, kinetics and biomarkers. Handb Exp Pharmacol2009; (192): 29-60.
13.
Benowitz NLCigarette smoking and cardiovascular disease: pathophysiology and implications for treatment. Prog Cardiovasc Dis2003; 46: 91-111.
14.
Zhu BQ, Heeschen C., Sievers RE, et al. Second hand smoke stimulates tumor angiogenesis and growth. Cancer Cell2003; 4: 191-196.
15.
Sikora L., Rao SP, Sriramarao P.Selectin-dependent rolling and adhesion of leukocytes in nicotine-exposed microvessels of lung allografts. Am J Physiol Lung Cell Mol Physiol2003; 285: L654-63.
16.
Puranik R., Celermajer DSSmoking and endothelial function. Prog Cardiovasc Dis2003; 45: 443-458.
17.
Heeschen C. , Jang JJ, Weis M., et al. Nicotine stimulates angiogenesis and promotes tumor growth and atherosclerosis. Nat Med2001; 7: 833-839.
18.
Heeschen C. , Weis M., Aicher A., Dimmeler S., Cooke J.A novel angiogenic pathway mediated by non-neuronal nicotinic acetylcholine receptors. J Clin Invest2002; 110: 527-536.
19.
Joseph AM, Norman SM, Ferry LH, et al. The safety of transdermal nicotine as an aid to smoking cessation in patients with cardiac disease. N Engl J Med1996; 335: 1792-1798.
Gotti C., Clementi F.Neuronal nicotinic receptors: from structure to pathology. Prog Neurobiol2004; 74: 363-396.
22.
Benhammou K. , Lee M., Strook M., et al. [(3)H]Nicotine binding in peripheral blood cells of smokers is correlated with the number of cigarettes smoked per day. Neuropharmacology2000; 39: 2818-2829.
23.
Lebargy F., Benhammou K., Morin D., et al. Tobacco smoking induces expression of very-high-affinity nicotine binding sites on blood polymorphonuclear cells. Am J Respir Crit Care Med1996; 153: 1056-1063.
24.
Moore GL, Ledford ME, Merydith A.A micromodification of the Drabkin hemoglobin assay for measuring plasma hemoglobin in the range of 5 to 2000 mg/dl. Biochem Med1981; 26: 167-173.
Forsberg EC , Prohaska SS, Katzman S., et al. Differential expression of novel potential regulators in hematopoietic stem cells. PLoS Genet2005; 1: e28.
27.
Christensen JL, Weissman ILFlk-2 is a marker in hematopoietic stem cell differentiation: a simple method to isolate long-term stem cells . Proc Natl Acad Sci U S A2001; 98: 14,541-14,546.
28.
Baum CM, Weissman IL, Tsukamoto AS, Buckle AM, Peault B.Isolation of a candidate human hematopoietic stem-cell population . Proc Natl Acad Sci U S A1992; 89: 2804-2808.
29.
Sabban EL, Gueorguiev VDEffects of short- and long-term nicotine treatment on intracellular calcium and tyrosine hydroxylase gene expression. Ann N Y Acad Sci2002; 971: 39-44.
30.
Spangrude GJ , Heimfeld S., Weissman ILPurification and characterization of mouse hematopoietic stem cells. Science1988; 241: 58-62.
31.
Spangrude GJ , Smith L., Uchida N., et al. Mouse hematopoietic stem cells. Blood1991; 78: 1395-1402.
32.
Wang Y., Pereira EFR, Maus ADJ, et al. Human bronchial epithelial and endothelial cells express alpha-7 nicotinic acetylcholine receptors. Mol Pharmacol2001; 60: 1201-1209.
33.
Benowitz NL , Zevin S., Jacob PRSources of variability in nicotine and cotinine levels with use of nicotine nasal spray, transdermal nicotine, and cigarette smoking. Br J Clin Pharmacol1997; 43: 259-267.
34.
Seggewiss R. , Buss EC, Herrmann D., Goldschmidt H., Ho AD, Fruehauf S.Kinetics of peripheral blood stem cell mobilization following G-CSF-supported chemotherapy . Stem Cells2003; 21: 568-574.
35.
Müller-Sieburg CE, Riblet R.Genetic control of the frequency of hematopoietic stem cells in mice: mapping of a candidate locus to chromosome 1. J Exp Med1996; 183: 1141-50.
36.
Ziche M., Morbidelli L., Choudhuri R., et al. Nitric oxide synthase lies downstream from vascular endothelial growth factor-induced but not basic fibroblast growth factor-induced angiogenesis . J Clin Invest1997; 99: 2625-2634.
37.
Ziche M.Nitric oxide promotes proliferation and plasminogen activator production by coronary venular endothelium through endogenous. Circ Res1993; 80: 845-852.
38.
Witzenbichler B., Asahara T., Murohara T., et al. Vascular endothelial growth factor-C (VEGF-C/VEGF-2) promotes angiogenesis in the setting of tissue ischemia. Am J Pathol1998; 153: 381-394.
39.
Noiri E., Lee E., Testa J., et al. Podokinesis in endothelial cell migration: role of nitric oxide . Am J Physiol1998; 274: C236-C244.
40.
Morbidelli L., Chang CH, Douglas JG, Granger HJ, Ledda F., Ziche M.Nitric oxide mediates mitogenic effect of VEGF on coronary venular endothelium. Am J Physiol1996; 270: H411-415.
41.
Lyden D., Hattori K., Dias S., et al. Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med2001; 7: 1194-1201.
42.
Inoue N., Venema RC, Sayegh HS, Ohara Y., Murphy TJ, Harrison DGMolecular regulation of the bovine endothelial cell nitric oxide synthase by transforming growth factor-beta 1. Arterioscler Thromb Vasc Biol1995; 15: 1255-1261.
43.
Heissig B., Hattori K., Dias S., et al. Recruitment of stem and progenitor cells from the bone marrow niche requires MMP-9 mediated release of kit-ligand. Cell2002; 109: 625-637.
44.
Koval LM, Zverkova AS, Grailhe R., et al. Nicotinic acetylcholine receptors alpha4beta2 and alpha7 regulate myeloand erythropoiesis within the bone marrow. Int J Biochem Cell Biol2008; 40: 980-990.
45.
Pandit TS, Sikora L., Muralidhar G., Rao SP, Sriramarao P.Sustained exposure to nicotine leads to extramedullary hematopoiesis in the spleen. Stem Cells2006; 24: 2373-2381.
46.
Serobyan N. , Schraufstatter IU, Strongin A., Khaldoyanidi SKNicotinic acetylcholine receptor-mediated stimulation of endothelial cells results in the arrest of haematopoietic progenitor cells on endothelium. Br J Haematol2005; 129: 257-265.
47.
Heeschen C. , Chang E., Aicher A., Cooke JPEndothelial progenitor cells participate in nicotine-mediated angiogenesis. J Am Coll Cardiol2006; 48: 255-360.
48.
Ernst E.Haemorheological consequences of chronic cigarette smoking. J Cardiovasc Risk1995; 2: 435-439.
49.
Cormier A., Paas Y., Zini R., et al. Long-term exposure to nicotine modulates the level and activity of acetylcholine receptors in white blood cells of smokers and model mice . Mol Pharmacol. 2004; 66: 1712-1718.
50.
Kimura R., Ushiyama N., Fujii T., Kawashima K.Nicotine-induced Ca2+ signaling and down-regulation of nicotinic acetylcholine receptor subunit alpha-expression in the CEM human leukemic T-cell line. Life Sci2003; 72: 2155-2158.
51.
Khaldoyanidi S., Sikora L., Orlovskaya I., Matrosova V., Kozlov V., Sriramarao P.Correlation between nicotine-induced inhibition of hematopoiesis and decreased CD44 expression on bone marrow stromal cells . Blood2001 ; 98: 303-312.
52.
Takahashi M. , Shigeno N., Narita M., et al. Cotinine (a metabolite of nicotine) suppresses the growth of hematopoietic progenitor cells at the concentration range equivalent to its serum levels in smokers . Stem Cells1999; 17: 125-126.
53.
Villablanca ACNicotine stimulates DNA synthesis and proliferation in vascular endothelial cells in vitro. J Appl Physiol1998; 84: 2089-2098.
Serobyan N. , Jagannathan S., Orlovskaya I., et al. The cholinergic system is involved in regulation of the development of the hematopoietic system. Life Sci2007 ; 80: 2352-2360.
56.
Rafii S., Lyden D.Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med2003; 9: 702-712.
57.
Aicher A., Heeschen C., Mildner-Rihm C., et al. Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med2003 ; 9: 1370-1376.
58.
Natori T., Sata M., Washida M., Hirata Y., Nagai R., Makuuchi M.Nicotine enhances neovascularization and promotes tumor growth. Mol Cells2003; 16: 143-146.
59.
Carty CS, Soloway PD, Kayastha S., et al. Nicotine and cotinine stimulate secretion of basic fibroblast growth factor and affect expression of matrix metalloproteinases in cultured human smooth muscle cells. J Vasc Surg1996 ; 24: 927-934.
60.
Conklin BS, Zhao W., Zhong DS, Chen C.Nicotine and cotinine up-regulate vascular endothelial growth factor expression in endothelial cells. Am J Pathol2002; 160: 413-418.
61.
Pelosi E., Valtieri M., Coppola S., et al. Identification of the hemangioblast in postnatal life. Blood2002; 100: 3203-3208.
62.
Kondo T., Hayashi M., Takeshita K., Numaguchi Y., Kobayashi K.Smoking cessation rapidly increases circulating progenitor cells in peripheral blood in chronic smokers. Arterioscler Thromb Vasc Biol2004; 24: 1442-1447.
63.
Czechowicz A., Kraft D., Weissman IL, Bhattacharya D.Efficient transplantation via antibody-based clearance of hematopoietic stem cell niches . Science2007; 318: 1296-1299.
64.
Wilson A., Trumpp A.Bone-marrow haematopoietic-stem-cell niches. Nat Rev Immunol2006; 6: 93-106.
