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Veterinary Research Communications

, Volume 37, Issue 4, pp 333–338 | Cite as

Apoptotic effects of tamoxifen on leukocytes from horse peripheral blood and bronchoalveolar lavage fluid

  • J. Sarmiento
  • B. Perez
  • N. Morales
  • C. Henriquez
  • L. Vidal
  • H. Folch
  • J. S. Galecio
  • G. MoránEmail author
Short Communication

Abstract

A reduction in inflammatory cell apoptosis is an important concept in the maintenance of inflammation and a potential target for the resolution of inflammation in many inflammatory diseases. Dysregulation of apoptosis has been implicated in a range of diseases, including tumors, neurodegenerative disorders and autoimmunity, and may also be implicated in allergic asthma. In horses, recurrent airway obstruction (RAO) is an asthma-like condition that is characterized increased survival neutrophil bronchial. Tamoxifen is a synthetic, non-steroidal, anti-estrogen agent that is widely used for treating all stages of breast cancer and has been approved for the prevention of breast cancer in high-risk women. The observed efficacy of tamoxifen has been attributed to both growth arrest and the induction of apoptosis. Therefore, the aim of our study was to evaluate the ability of tamoxifen to induce apoptosis in vitro in granulocytic cells from peripheral blood and in mononuclear cells from bronchoalveolar lavage fluid (BALF) in horses. Flow cytometry using commercial AnnexinV-FITC and propidium iodide was used to quantify early and late apoptotic leukocytes, respectively. The results showed a significant increase in early apoptosis in peripheral blood and bronchial granulocytic cells treated with tamoxifen. The rate of early apoptosis of mononuclear cells from blood and BALF when incubated with tamoxifen was significantly lower compared with granulocytic cells. We did not observe a direct effect of tamoxifen on late apoptosis in any of the in vitro assays in the cell types used here. These results indicate that the apoptotic mechanisms under these experimental conditions would affect only blood and BALF granulocytic cells, particularly in early apoptosis. Finally, further in vitro and in vivo studies are needed to better understand apoptotic mechanisms because tamoxifen could be used to treat chronic, inflammatory pathologies associated with granulocytes and allergic diseases, such as asthma or equine RAO.

Keywords

Apoptosis Tamoxifen Horse 

Abbreviations

BALF

bronchoalveolar lavage fluid

FCS

fetal calves serum

PI

propidium iodide

RAO

recurrent airway obstruction

Notes

Acknowledgments

This work was supported by FONDECYT Nº 1130355 (Conicyt- Chilean Government).

Conflicts of interest

None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper.

References

  1. Ahn SJ, Yoon MS, Hyuk S, Han W, Yoon YD, Han JS, Noh DY (2003) Phospholipase C-protein kinase C mediated phospholipase D activation pathway is involved in tamoxifen induced apoptosis. J Cell Biochem 89:520–528PubMedCrossRefGoogle Scholar
  2. Ainsworth DM, Grünig G, Matychak MB, Young J, Wagner B, Erb HN, Antczak DF (2003) Recurrent airway obstruction (RAO) in horses is characterized by IFN-g and IL-8 production in bronchoalveolar lavage cells. Vet Immunol Immunopathol 96:83–91PubMedCrossRefGoogle Scholar
  3. Ainsworth DM, Wagner B, Franchini M, Grunig G, Erb H, Tan JY (2006) Time-dependent alterations in gene expression of interleukin-8 in the bronchial epithelium of horses with recurrent airway obstruction. Am J Vet Res 67:669–677PubMedCrossRefGoogle Scholar
  4. Ainsworth DM, Wagner B, Erb HN, Young JC, Retallick DE (2007) Effects of in vitro exposure to hay dust on expression of interleukin-17, −23, −8, and −1β and chemokine (C-X-C motif) ligand 2 by pulmonary mononuclear cells isolated from horses chronically affected with recurrent airway disease. Am J Vet Res 68:1361–1369PubMedCrossRefGoogle Scholar
  5. Akbar AN, Salmon M (1997) Cellular environments and apoptosis: tissue microenvironments control activated T cell death. Immunol Today 18:72–76PubMedCrossRefGoogle Scholar
  6. Babina M, Kirn F, Hoser D, Ernst D, Rohde W, Zuberbier T, Worm M (2010) Tamoxifen counteracts the allergic immune response and improves allergen-induced dermatitis in mice. Clin Exp Allergy 40:1256–1265PubMedCrossRefGoogle Scholar
  7. Breuer J, Mueller U, Locher L, Spallek A, Recknagel S, Uhlig A, Schusser GF (2011) Differentiation of viable, apoptotic and necrotic cells in bronchoalveolar lavage fluid of normal horses and horses with recurrent airway obstruction. Berliner und Muenchener Tieraerztliche Wochenschrift 124:154–160Google Scholar
  8. Budtz PE (1999) Role of proliferation and apoptosis in net growth rates of human breast cancer cells (MCF-7) treated with oestradiol and/or tamoxifen. Cell Prolif 32:289–302PubMedCrossRefGoogle Scholar
  9. Cabot MC, Zhang Z, Cao H, Lavie Y, Giuliano AE, Han TY, Jones RC (1997) Tamoxifen activates cellular phospholipase C and D and elicits protein kinase C translocation. Int J Cancer 70:567–574PubMedCrossRefGoogle Scholar
  10. Cameron DA, Keen JC, Dixon JM, Bellamy C, Hanby A, Anderson TJ, Miller WR (2000) Effective tamoxifen therapy of breast cancer involves both antiproliferative and pro-apoptotic changes. Eur J Cancer 36:845–851PubMedCrossRefGoogle Scholar
  11. Cohen JJ (1999) Apoptosis: mechanisms of life and death in the immune system. J Allergy Clin Immunol 103:548–554PubMedCrossRefGoogle Scholar
  12. Cordeau ME, Joubert P, Dewachi O, Hamid Q, Lavoie JP (2004) IL-4, IL-5 and IFN-g mRNA expression in pulmonary lymphocytes in equine heaves. Vet Immunolo Immunopathol 97:87–96CrossRefGoogle Scholar
  13. Cormica L, O’Sullivan S, Burke CM, Poulter LW (2001) IFNgamma but not IL-4 T cells of the asthmatic bronchial wall show increased incidence of apoptosis. Clin Exp Allergy 31:731–739CrossRefGoogle Scholar
  14. Curik I, Fraser D, Eder C, Achmann R, Swinburne J, Binns M, Crameri R, Brem G, Sölkner J, Marti E (2003) Association between MHC gene region and variation of serum IgE levels against specific mould allergens in the horse. Genet Select Evol 35:117–190Google Scholar
  15. Datta R, Kojima H, Yoshida K, Kufe D (1997) Caspase-3- mediated cleavage of protein kinase C theta in induction of apoptosis. J Biol Chem 272:20317–20320PubMedCrossRefGoogle Scholar
  16. De Rose V, Cappello P, Sorbello V, Ceccarini B, Gani F, Bosticardo M, Fassio S, Novelli F (2004) IFN-gamma inhibits the proliferation of allergen-activated T lymphocytes from atopic, asthmatic patients by inducing Fas/FasL-mediated apoptosis. J Leukoc Biol 76:423–432PubMedCrossRefGoogle Scholar
  17. Debrue M, Hamilton E, Joubert P, Lajoie-Kadoch S, Lavoie JP (2005) Chronic exacerbation of equine heaves is associated with an increased expression of interleukin-17 mRNA in bronchoalveolar lavage cells. Vet Immunol Immunopathol 105:25–31PubMedCrossRefGoogle Scholar
  18. Dibbert B, Weber M, Nikolaizik WH, Vogt P, Schöni MH, Blaser K, Simon HU (1999) Cytokine-mediated Bax deficiency and consequent delayed neutrophil apoptosis: a general mechanism to accumulate effector cells in inflammation. Proc Natl Acad Sci 96:13330–13335PubMedCrossRefGoogle Scholar
  19. Duffy SM, Lawley WJ, Kaur D, Yang W, Bradding P (2003) Inhibition of human mast cell proliferation and survival by tamoxifen in association with ion channel modulation. J Allergy Clin Immunol 112:965–972PubMedCrossRefGoogle Scholar
  20. Eder C, Crameri R, Mayer C, Eicher R, Straub R, Gerber H, Lazary S, Marti E (2000) Allergen-specific IgE levels against crude mould and storage mite extracts and recombinant mould allergens in sera from horses affected with chronic bronchitis. Vet Immunol Immunopathol 73:241–253PubMedCrossRefGoogle Scholar
  21. Eder C, Curik I, Brem G, Crameri R, Bodo I, Habe F, Lazary S, Sölkner J, Marti E (2001) Influence of environmental and genetic factors on allergen-specific immunoglobulin E levels in sera from Lipizzan horses. Equine Vet J 33:714–720PubMedCrossRefGoogle Scholar
  22. Finotto S, Eigenbrod T, Karwot R, Boross I, Doganci A, Ito H, Nishimoto N, Yoshizaki K, Kishimoto T, Rose-John S, Galle PR, Neurath MF (2007) Local blockade of IL-6R signaling induces lung CD4+ T cell apoptosis in a murine model of asthma via regulatory T cells. Int Immunol 19:685–693PubMedCrossRefGoogle Scholar
  23. Franchini M, Gill U, Von Fellenberg R, Bracher VV (2000) Interleukin-8 concentration and neutrophil chemotactic activity in bronchoalveolar lavage fluid of horses with chronic obstructive pulmonary disease following exposure to hay. Am J Vet Res 11:1369–1374CrossRefGoogle Scholar
  24. Gerber V, Lindberg A, Berney C, Robinson NE (2004) Airway mucus in recurrent airway obstruction—short-term response to environmental challenge. J Vet Inter Med 18:92–97Google Scholar
  25. Giguere S, Viel L, Lee E, MacKay RJ, Hernandez J, Franchini M (2002) Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate. Vet Immunol Immunopathol 85:147–158PubMedCrossRefGoogle Scholar
  26. Halliwell REW, McGorum BC, Irving P, Dixon PM (1993) Local and systemic antibody production in horses affected with chronic obstructive pulmonary disease. Vet Immunol Immunopathol 38:201–205PubMedCrossRefGoogle Scholar
  27. Haslett C (1999) Granulocyte apoptosis and its role in the resolution and control of lung inflammation. Am J Respir Crit Care Med 160:S5–S11PubMedCrossRefGoogle Scholar
  28. Jayaraman S, Castro M, O’Sullivan M, Bragdon MJ, Holtzman MJ (1999) Resistance to Fas-mediated T cell apoptosis in asthma. J Immunol 162:1717–1722PubMedGoogle Scholar
  29. Kisanga ER, Gjerde J, Guerrieri-Gonzaga A, Pigatto F, Pesci-Feltri A, Robertson C, Serrano D, Pelosi G, Decensi A, Lien EA (2004) Tamoxifen and metabolite concentrations in serum and breast cancer tissue during three dose regimens in a randomized preoperative trial. Clin Cancer Res 10:2336–2343PubMedCrossRefGoogle Scholar
  30. Künzle F, Gerber V, van der Haegen A, Wampfler B, Straub R, Marti E (2007) IgE-bearing cells in bronchoalveolar lavage fluid and allergen-specific IgE levels in sera from RAO-affected horses. J Vet Med A Physiol Pathol Clin Med 54:40–47PubMedCrossRefGoogle Scholar
  31. Lavoie JP, Maghni K, Desnoyers M, Taha R, Martin JG, Hamid Q (2001) Neutrophilic airway inflammation in horses with heaves is characterized by a Th2-type cytokine profile. Am J Respir Crit Care Med 164:1410–1413PubMedCrossRefGoogle Scholar
  32. Leguillette R (2003) Recurrent airway obstruction-heaves. Vet Clin North Am Equine Pract 19:63–68PubMedCrossRefGoogle Scholar
  33. Marchette LC, Marchette BE, Abraham WM, Wanner A (1985) The effect of systemic hydration on normal and impaired mucociliary function. Pediatr Pulmonol 1:107–111PubMedCrossRefGoogle Scholar
  34. Morán G, Burgos R, Araya O, Folch H (2010a) In vitro bioassay to detect reaginic antibodies from the serum of horses affected with recurrent airway obstruction. Vet Res Commun 34:91–99PubMedCrossRefGoogle Scholar
  35. Morán G, Folch H, Burgos R, Araya O, Barria M (2010b) Detection of reaginic antibodies against Faenia rectivirgula from the serum of horses affected with recurrent airway obstruction by an in vitro bioassay. Vet Res Commun 34:719–726PubMedCrossRefGoogle Scholar
  36. Moran G, Buechner-Maxwell VA, Folch H, Henriquez C, Galecio JS, Perez B, Carrasco C, Barria A (2011) Increased apoptosis of CD4 and CD8 T lymphocytes in the airways of horses with recurrent airway obstruction. Vet Res Commun 35:447–456PubMedCrossRefGoogle Scholar
  37. Moran G, Perez B, Morales N, Vidal L, Galecio JS, Vasquez N, Folch H, Henriquez C (2012) Apoptotic effect of Tamoxifen on neutrophils bronchial cells in horses with recurrent airway obstruction: a new therapeutic approach. Immunology (Supp) 137:438–439Google Scholar
  38. Müller M, Grunewald J, Olgart-Höglund C, Dahlén B, Eklund A, Stridh H (2006) Altered apoptosis in bronchoalveolar lavage lymphocytes after allergen exposure of atopic asthmatic subjects. Eur Respir J 28:513–522PubMedCrossRefGoogle Scholar
  39. O’Brian CA, Ward NE, Anderson BW (1988) Role of specific interactions between protein kinase C and triphenylethylenes in inhibition of the enzyme. J Natl Cancer Inst 80:1628–1633PubMedCrossRefGoogle Scholar
  40. Riihimäki M, Raine A, Art T, Lekeux P, Couëtil L, Pringle J (2008) Partial divergence of cytokine mRNA expression bronchial tissues compared to broncheoalveolar lavage cells in horses with recurrent airway obstruction. Vet Immunol Immunopathol 122:256–264PubMedCrossRefGoogle Scholar
  41. Robinson NE (2001) International workshop on equine chronic airway disease Michigan State University. Equine Vet J 33:5–19PubMedCrossRefGoogle Scholar
  42. Schmallenbach KH, Rahman I, Sasse HH, Dixon PM, Halliwell RE, McGorum BC, Miller HR (1998) Studies on pulmonary and systemic Aspergillus Fumigatus-specific IgE and IgG antibodies in horses affected with chronic obstructive pulmonary disease (COPD). Vet Immunol Immunopathol 66:245–256PubMedCrossRefGoogle Scholar
  43. Spinozzi F, Fizzotti M, Agea E, Piattoni S, Droetto S, Russano A, Forenza N, Bassotti G, Grignani F, Bertotto A (1998) Defective expression of Fas messenger RNA and FAS receptor on pulmonary T cells from patients with asthma. Ann Intern Med 128:363–369PubMedCrossRefGoogle Scholar
  44. Stearns V, Johnson MD, Rae JM, Morocho A, Novielli A, Bhargava P, Hayes DF, Desta Z, Flockhart DA (2003) Active tamoxifen metabolite plasma concentrations after coadministration of tamoxifen and the selective serotonin reuptake inhibitor paroxetine. J Natl Cancer Inst 95:1758–1764PubMedCrossRefGoogle Scholar
  45. Tahon L, Baselgia S, Gerber V, Doherr MG, Straub R, Robinson NE, Marti E (2009) In vitro allergy test compared to intradermal testing in horses with recurrent airway obstruction. Vet Immunol Immunopathol 127:85–93PubMedCrossRefGoogle Scholar
  46. Thompson CB (1995) Apoptosis in the pathogenesis and treatment of disease. Science 267:1456–1462PubMedCrossRefGoogle Scholar
  47. Tong J, Bandulwala HS, Clay BS, Anders RA, Shilling RA, Balachandran DD, Chen B, Weinstock JV, Solway J, Hamann KJ, Sperling AI (2006) Fas-positive T cells regulate the resolution of airway inflammation in a murine model of asthma. J Exp Med 203:1173–1184PubMedCrossRefGoogle Scholar
  48. Turlej RK, Fievez L, Sandersen CF, Dogné S, Kirschvink N, Lekeux P, Bureau F (2001) Enhanced survival of lung granulocytes in an animal model of asthma: evidence for a role of GM-CSF activated STAT5 signaling pathway. Thorax 56:696–702PubMedCrossRefGoogle Scholar
  49. Vignola AM, Chanez P, Chiappara G, Siena L, Merendino A, Reina C, Gagliardo R, Profita M, Bousquet J, Bonsignore G (1999) Evaluation of apoptosis of eosinophils, macrophages, and T lymphocytes in mucosal biopsy specimens of patients with asthma and chronic bronchitis. J Allergy Clin Immunol 103:563–573PubMedCrossRefGoogle Scholar
  50. Zizzo G, Cohen PL (2013) IL-17 stimulates differentiation of human anti-inflammatory macrophages and phagocytosis of apoptotic neutrophils in response to IL-10 and glucocorticoids. J Immunol 190:5237–5246PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • J. Sarmiento
    • 1
  • B. Perez
    • 2
  • N. Morales
    • 2
  • C. Henriquez
    • 3
  • L. Vidal
    • 2
  • H. Folch
    • 3
  • J. S. Galecio
    • 4
  • G. Morán
    • 2
    Email author
  1. 1.Department of Phisiology, Faculty of MedicineUniversidad Austral de ChileValdiviaChile
  2. 2.Department of Pharmacology, Faculty of Veterinary ScienceUniversidad Austral de ChileValdiviaChile
  3. 3.Department of Immunology, Faculty of MedicineUniversidad Austral de ChileValdiviaChile
  4. 4.Department of Clinical Veterinary Science, Faculty of Veterinary ScienceUniversidad Austral de ChileValdiviaChile

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