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Comparative study of topical anti-allergic eye drops on human conjunctiva-derived cells: responses to histamine and IFNγ and toxicological profiles

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Abstract

Background

The purpose of the study was to compare toxic effects and responses to histamine and IFNγ associated with the use of some widely used anti-allergic eye drops commercially available today.

Methods

For dynamic studies, the Wong-Kilbourne cell line was stimulated for 24 h with histamine or IFNγ in the presence or absence of anti-allergic eye drops. Supernatants of histamine-stimulated cells were evaluated for the production of IL-6 and IL-8 by ELISA, while the expression of ICAM-1 was evaluated by flow cytometry on IFNγ-stimulated cells. Toxicological assays were performed using cold light cytofluorometry: viability and apoptosis as well as reactive oxygen species (ROS) and O2.- production were assessed using neutral red, Hoechst/propidium iodide, H2-DCFDA and hydroethidine tests, respectively.

Results

Antihistamines reduced IL-6 release and presented dose-dependent inhibitory effects on IL-8 production. None of the eye drops decreased the basal or IFNγ-stimulated expression of ICAM-1. Conversely, eye drops preserved with benzalkonium chloride (BAC) induced even higher ICAM-1 expression levels on IFNγ-stimulated cells than did IFNγ alone, whereas unpreserved drugs had no effect. Toxicological assays confirmed the pivotal role of BAC in proportionally reducing cell viability while increasing apoptosis and oxidative stress.

Conclusions

The ability of topical ocular anti-H1 drugs to significantly reduce the production of IL-6 and IL-8 argues that they may help treat the inflammatory processes occurring in allergic ocular surface disorders. Nevertheless, preserved ophthalmic formulations may enhance epithelial conjunctival expression of ICAM-1 in the presence of a low inflammatory stimulus, such as IFNγ, and displayed toxic as well as pro-oxidative effects on these cells. Therefore, BAC used as preservative might in part interfere with the potential anti-inflammatory properties of the active compound by modulating the immuno-inflammatory response of epithelial conjunctival cells.

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References

  1. Abelson MB, Chambers WA, Smith LM (1990) Conjunctival allergen challenge. A clinical approach to studying allergic conjunctivitis. Arch Ophthalmol 108:84–88

    PubMed  CAS  Google Scholar 

  2. Abelson MB, Schaefer K (1993) Conjunctivitis of allergic origin: immunologic mechanisms and current approaches to therapy. Surv Ophthalmol 38(Suppl):115–132

    Article  PubMed  Google Scholar 

  3. Abelson MB, Loeffler O (2003) Conjunctival allergen challenge: models in the investigation of ocular allergy. Curr Allergy Asthma Rep 3:363–368

    PubMed  Google Scholar 

  4. Abelson MB (2004) A review of olopatadine for the treatment of ocular allergy. Exp Opin Pharmacother 5:1979–1994

    Article  CAS  Google Scholar 

  5. Abelson MB, Gomes P, Crampton HJ, Schiffman RM, Bradford RR, Whitcup SM (2004) Efficacy and tolerability of ophthalmic epinastine assessed using the conjunctival antigen challenge model in patients with a history of allergic conjunctivitis. Clin Ther 26:35–47

    Article  PubMed  CAS  Google Scholar 

  6. Bachert C (1998) Histamine-a major role in allergy? Clin Exp Allergy 28(Suppl 6):15–19

    Article  PubMed  CAS  Google Scholar 

  7. Bagnasco M, Pesce G, Fiorino N, Riccio AM, Ciprandi G, Buscaglia S, Canonica GW (1997) In situ hybridization analysis of ICAM-1 (CD54) mRNA on conjunctival epithelium during allergic inflammation. Clin Exp Allergy 27:737–743

    Article  PubMed  CAS  Google Scholar 

  8. Baudouin C, Hamard P, Liang H, Creuzot-Garcher C, Bensoussan L, Brignole F (2004) Conjunctival epithelial cell expression of interleukins and inflammatory markers in glaucoma patients treated over the long term. Ophthalmology 111:2186–2192

    Article  PubMed  Google Scholar 

  9. Baudouin C, Liang H, Bremond-Gignac D, Hamard P, Hreiche R, Creuzot-Garcher C, Warnet JM, Brignole-Baudouin F (2005) CCR 4 and CCR 5 expression in conjunctival specimens as differential markers of T(H)1/ T(H)2 in ocular surface disorders. J Allergy Clin Immunol 116:614–619

    Article  PubMed  CAS  Google Scholar 

  10. Belloc F, Dumain P, Boisseau MR, Jalloustre C, Reiffers J, Bernard P, Lacombe F (1994) A flow cytometric method using Hoechst 33342 and propidium iodide for simultaneous cell cycle analysis and apoptosis determination in unfixed cells. Cytometry 17:59–65

    Article  PubMed  CAS  Google Scholar 

  11. Brignole-Baudouin F, Ott AC, Warnet JM, Baudouin C (2004) Flow cytometry in conjunctival impression cytology: a new tool for exploring ocular surface pathologies. Exp Eye Res 78:473–481

    Article  PubMed  CAS  Google Scholar 

  12. Buscaglia S, Paolieri F, Catrullo A, Fiorino N, Riccio AM, Pesce G, Montagna P, Bagnasco M, Ciprandi G, Canonica GW (1996) Topical ocular levocabastine reduces ICAM-1 expression on epithelial cells both in vivo and in vitro. Clin Exp Allergy 26:1188–1196

    Article  PubMed  CAS  Google Scholar 

  13. Calder VL, Jolly G, Hingorani M, Adamson P, Leonardi A, Secchi AG, Buckley RJ, Lightman S (1999) Cytokine production and mRNA expression by conjunctival T-cell lines in chronic allergic eye disease. Clin Exp Allergy 29:1214–1222

    Article  PubMed  CAS  Google Scholar 

  14. Calderon E, Lockey RF (1992) A possible role for adhesion molecules in asthma. J Allergy Clin Immunol 90:852–865

    Article  PubMed  CAS  Google Scholar 

  15. Canonica GW, Ciprandi G, Buscaglia S, Pesce G, Bagnasco M (1994) Adhesion molecules of allergic inflammation: recent insights into their functional roles. Allergy 49:135–141

    PubMed  CAS  Google Scholar 

  16. Canonica GW, Ciprandi G, Passalacqua G, Pesce G, Scordamaglia A, Bagnasco M (1997) Molecular events in allergic inflammation: experimental models and possible modulation. Allergy 52:25–30

    Article  PubMed  CAS  Google Scholar 

  17. Canonica GW (1997) Adhesion molecules in allergy. J Invest Allergol Clin Immunol 7:274–276

    CAS  Google Scholar 

  18. Canonica GW (1997) Adhesion molecules and H1-antihistamines. Clin Exp Allergy 27(Suppl 2):19–27; discussion 54–56

    Article  PubMed  CAS  Google Scholar 

  19. Ciprandi G, Buscaglia S, Pesce G, Villaggio B, Bagnasco M, Canonica GW (1993) Allergic subjects express intercellular adhesion molecule-1 (ICAM-1 or CD54) on epithelial cells of conjunctiva after allergen challenge. J Allergy Clin Immunol 91:783–792

    Article  PubMed  CAS  Google Scholar 

  20. Ciprandi G, Pronzato C, Ricca V, Passalacqua G, Bagnasco M, Canonica GW (1994) Allergen-specific challenge induces intercellular adhesion molecule 1 (ICAM-1 or CD54) on nasal epithelial cells in allergic subjects. Relationships with early and late inflammatory phenomena. Am J Respir Crit Care Med 150:1653–1659

    PubMed  CAS  Google Scholar 

  21. Ciprandi G, Pronzato C, Passalacqua G, Ricca V, Grogen J, Mela GS, Varese P, Bertolini C, Bagnasco M, Canonica GW (1996) Topical azelastine reduces eosinophil activation and intercellular adhesion molecule-1 expression on nasal epithelial cells: an antiallergic activity. J Allergy Clin Immunol 98:1088–1096

    Article  PubMed  CAS  Google Scholar 

  22. Ciprandi G, Cosentino C, Milanese M, Tosca MA (2003) Rapid anti-inflammatory action of azelastine eyedrops for ongoing allergic reactions. Ann Allergy Asthma Immunol 90:434–438

    PubMed  CAS  Google Scholar 

  23. Ciprandi G, Tosca MA, Cosentino C, Riccio AM, Passalacqua G, Canonica GW (2003) Effects of fexofenadine and other antihistamines on components of the allergic response: adhesion molecules. J Allergy Clin Immunol 112:S78–S82

    Article  PubMed  CAS  Google Scholar 

  24. Cook EB, Stahl JL, Barney NP, Graziano FM (2001) Olopatadine inhibits anti-immunoglobulin E-stimulated conjunctival mast cell upregulation of ICAM-1 expression on conjunctival epithelial cells. Ann Allergy Asthma Immunol 87:424–429

    PubMed  CAS  Google Scholar 

  25. Cvenkel B, Ihan A (2002) Ocular surface changes induced by topical antiglaucoma monotherapy. Ophthalmologica 216:175–179

    Article  PubMed  CAS  Google Scholar 

  26. Debbasch C, Pisella PJ, Rat P, Warnet JM, Baudouin C (2001) Antioxidant effects of mast cell inhibitors in a human conjunctival cell line. J Fr Ophtalmol 24:121–128

    PubMed  CAS  Google Scholar 

  27. Debbasch C, Brignole F, Pisella PJ, Warnet JM, Rat P, Baudouin C (2001) Quaternary ammoniums and other preservatives’ contribution in oxidative stress and apoptosis on Chang conjunctival cells. Invest Ophthalmol Vis Sci 42:642–652

    PubMed  CAS  Google Scholar 

  28. Debbasch C, Pisella PJ, De Saint Jean M, Rat P, Warnet JM, Baudouin C (2001) Mitochondrial activity and glutathione injury in apoptosis induced by unpreserved and preserved beta-blockers on Chang conjunctival cells. Invest Ophthalmol Vis Sci 42:2525–2533

    PubMed  CAS  Google Scholar 

  29. Debbasch C, De La Salle SB, Brignole F, Rat P, Warnet JM, Baudouin C (2002) Cytoprotective effects of hyaluronic acid and Carbomer 934P in ocular surface epithelial cells. Invest Ophthalmol Vis Sci 43:3409–3415

    PubMed  Google Scholar 

  30. De Saint Jean M, Brignole F, Feldmann G, Goguel A, Baudouin C (1999) Interferon-gamma induces apoptosis and expression of inflammation-related proteins in Chang conjunctival cells. Invest Ophthalmol Vis Sci 40:2199–2212

    PubMed  Google Scholar 

  31. De Saint Jean M, Brignole F, Bringuier AF, Bauchet A, Feldmann G, Baudouin C (1999) Effects of benzalkonium chloride on growth and survival of Chang conjunctival cells. Invest Ophthalmol Vis Sci 40:619–630

    PubMed  Google Scholar 

  32. De Saint Jean M, Debbasch C, Brignole F, Rat P, Warnet JM, Baudouin C (2000) Toxicity of preserved and unpreserved antiglaucoma topical drugs in an in vitro model of conjunctival cells. Curr Eye Res 20:85–94

    Article  PubMed  Google Scholar 

  33. De Saint Jean M, Baudouin C, Di Nolfo M, Roman S, Lozato P, Warnet JM, Brignole F (2004) Comparison of morphological and functional characteristics of primary-cultured human conjunctival epithelium and of Wong-Kilbourne derivative of Chang conjunctival cell line. Exp Eye Res 78:257–274

    Article  PubMed  CAS  Google Scholar 

  34. Fujihara T, Takeuchi T, Saito K, Tsubota K (1997) Flow cytometric analysis of surface antigens on human conjunctival epithelial cells. Ophthalm Res 29:103–109

    Article  CAS  Google Scholar 

  35. Gamache DA, Dimitrijevich SD, Weimer LK, Lang LS, Spellman JM, Graff G, Yanni JM (1997) Secretion of proinflammatory cytokines by human conjunctival epithelial cells. Ocul Immunol Inflamm 5:117–128

    PubMed  CAS  Google Scholar 

  36. Garrelds IM, de Graaf-in’t Veld T, van Toorenenbergen AW, van Wijk RG, Boegheim JP, Saxena PR, Zijlstra FJ (1996) Selective effect of levocabastine on histamine receptor and histamine release from human leukocytes and guinea pig isolated tissue. Ann Allergy Asthma Immunol 76:327–334

    PubMed  CAS  Google Scholar 

  37. Guenoun JM, Baudouin C, Rat P, Pauly A, Warnet JM, Brignole-Baudouin F (2005) In vitro study of inflammatory potential and toxicity profile of latanoprost, travoprost, and bimatoprost in conjunctiva-derived epithelial cells. Invest Ophthalmol Vis Sci 46:2444–2450

    Article  PubMed  Google Scholar 

  38. Guenoun JM, Baudouin C, Rat P, Pauly A, Warnet JM, Brignole-Baudouin F (2005) In vitro comparison of cytoprotective and antioxidative effects of latanoprost, travoprost, and bimatoprost on conjunctiva-derived epithelial cells. Invest Ophthalmol Vis Sci 46:4594–4599

    Article  PubMed  Google Scholar 

  39. Guglielminetti E, Barabino S, Monaco M, Mantero S, Rolando M (2002) HLA-DR expression in conjunctival cells after latanoprost. J Ocul Pharmacol Ther 18:1–9

    Article  PubMed  CAS  Google Scholar 

  40. Hingorani M, Calder VL, Buckley RJ, Lightman SL (1998) The role of conjunctival epithelial cells in chronic ocular allergic disease. Exp Eye Res 67:491–500

    Article  PubMed  CAS  Google Scholar 

  41. Irkec M, Bozkurt B (2003) Epithelial cells in ocular allergy. Curr Allergy Asthma Rep 3:352–357

    PubMed  Google Scholar 

  42. Jambou D, Lapalus P (1990) Effect of N-acetyl-aspartyl-glutamate (Naaga) on in-vitro leukotriene synthesis by macrophage cell line P388D1. Int J Tissue React 12:273–280

    PubMed  CAS  Google Scholar 

  43. Leonardi A, DeFranchis G, Zancanaro F, Crivellari G, De Paoli M, Plebani M, Secchi AG (1999) Identification of local Th2 and Th0 lymphocytes in vernal conjunctivitis by cytokine flow cytometry. Invest Ophthalmol Vis Sci 40:3036–3040

    PubMed  CAS  Google Scholar 

  44. Leonardi A, DeFranchis G, De Paoli M, Fregona I, Plebani M, Secchi A (2002) Histamine-induced cytokine production and ICAM-1 expression in human conjunctival fibroblasts. Curr Eye Res 25:189–196

    Article  PubMed  Google Scholar 

  45. Martin AP, Urrets-Zavalia J, Berra A, Mariani AL, Gallino N, Gomez Demel E, Gagliardi J, Baena-Cagnani CE, Urrets-Zavalia E, Serra HM (2003) The effect of ketotifen on inflammatory markers in allergic conjunctivitis: an open, uncontrolled study. BMC Ophthalmol 3:2

    Article  PubMed  Google Scholar 

  46. Matsukura M, Yajima A, Yamazaki F, Yudate T, Yamada H, Tezuka T (2003) Epinastine inhibits eosinophil chemotaxis and adhesion molecules in atopic dermatitis. Skin Pharmacol Appl Skin Physiol 16:405–410

    Article  PubMed  CAS  Google Scholar 

  47. Metz DP, Hingorani M, Calder VL, Buckley RJ, Lightman SL (1997) T-cell cytokines in chronic allergic eye disease. J Allergy Clin Immunol 100:817–824

    Article  PubMed  CAS  Google Scholar 

  48. Miki I, Kusano A, Ohta S, Hanai N, Otoshi M, Masaki S, Sato S, Ohmori K (1996) Histamine enhanced the TNF-alpha-induced expression of E-selectin and ICAM-1 on vascular endothelial cells. Cell Immunol 171:285–288

    PubMed  CAS  Google Scholar 

  49. Montefort S, Holgate ST (1991) Adhesion molecules and their role in inflammation. Respir Med 85:91–99

    PubMed  CAS  Google Scholar 

  50. Paolieri F, Battifora M, Riccio AM, Pesce G, Canonica GW, Bagnasco M (1997) Intercellular adhesion molecule-1 on cultured human epithelial cell lines: influence of proinflammatory cytokines. Allergy 52:521–531

    PubMed  CAS  Google Scholar 

  51. Paolieri F, Battifora M, Riccio AM, Bertolini C, Cutolo M, Bloom M, Ciprandi G, Canonica GW, Bagnasco M (1998) Terfenadine and fexofenadine reduce in vitro ICAM-1 expression on human continuous cell lines. Ann Allergy Asthma Immunol 81:601–607

    PubMed  CAS  Google Scholar 

  52. Patalano F, Ruggieri F (1989) Sodium cromoglycate: a review. Eur Respir J Suppl 6:556s–560s

    PubMed  CAS  Google Scholar 

  53. Pisella PJ, Fillacier K, Elena PP, Debbasch C, Baudouin C (2000) Comparison of the effects of preserved and unpreserved formulations of timolol on the ocular surface of albino rabbits. Ophthalm Res 32:3–8

    Article  CAS  Google Scholar 

  54. Pisella PJ, Debbasch C, Hamard P, Creuzot-Garcher C, Rat P, Brignole F, Baudouin C (2004) Conjunctival proinflammatory and proapoptotic effects of latanoprost and preserved and unpreserved timolol: an ex vivo and in vitro study. Invest Ophthalmol Vis Sci 45:1360–1368

    Article  PubMed  Google Scholar 

  55. Schoch C (2003) Effects of ketotifen 0.025% and lodoxamide 0.1% on eosinophil infiltration into the guinea pig conjunctiva in a model of allergic conjunctivitis. J Ocul Pharmacol Ther 19:153–159

    Article  PubMed  CAS  Google Scholar 

  56. Sharif NA, Xu SX, Magnino PE, Pang IH (1996) Human conjunctival epithelial cells express histamine-1 receptors coupled to phosphoinositide turnover and intracellular calcium mobilization: role in ocular allergic and inflammatory diseases. Exp Eye Res 63:169–178

    Article  PubMed  CAS  Google Scholar 

  57. Smit EE, Sra SK, Grabowski LR, Ward SL, Trocme SD (2003) Modulation of IL-8 and RANTES release in human conjunctival epithelial cells: primary cells and cell line compared and contrasted. Cornea 22:332–337

    Article  PubMed  Google Scholar 

  58. Springer TA (1990) Adhesion receptors of the immune system. Nature 346:425–434

    Article  PubMed  CAS  Google Scholar 

  59. Stahl JL, Cook EB, Graziano FM, Barney NP (2003) Differential and cooperative effects of TNFalpha, IL-1beta, and IFNgamma on human conjunctival epithelial cell receptor expression and chemokine release. Invest Ophthalmol Vis Sci 44:2010–2015

    Article  PubMed  Google Scholar 

  60. Triggiani M, Gentile M, Secondo A, Granata F, Oriente A, Taglialatela M, Annunziato L, Marone G (2001) Histamine induces exocytosis and IL-6 production from human lung macrophages through interaction with H1 receptors. J Immunol 166:4083–4091

    PubMed  CAS  Google Scholar 

  61. Wegner CD, Gundel RH, Reilly P, Haynes N, Letts LG, Rothlein R (1990) Intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of asthma. Science 247:456–459

    Article  PubMed  CAS  Google Scholar 

  62. Weimer LK, Gamache DA, Yanni JM (1998) Histamine-stimulated cytokine secretion from human conjunctival epithelial cells: inhibition by the histamine H1 antagonist emedastine. Int Arch Allergy Immunol 115:288–293

    Article  PubMed  CAS  Google Scholar 

  63. White MV (1990) The role of histamine in allergic diseases. J Allergy Clin Immunol 86:599–605

    Article  PubMed  CAS  Google Scholar 

  64. Yanni JM, Miller ST, Gamache DA, Spellman JM, Xu S, Sharif NA (1997) Comparative effects of topical ocular anti-allergy drugs on human conjunctival mast cells. Ann Allergy Asthma Immunol 79:541–545

    Article  PubMed  CAS  Google Scholar 

  65. Yanni JM, Weimer LK, Sharif NA, Xu SX, Gamache DA, Spellman JM (1999) Inhibition of histamine-induced human conjunctival epithelial cell responses by ocular allergy drugs. Arch Ophthalmol 117:643–647

    PubMed  CAS  Google Scholar 

  66. Zhan H, Towler HM, Calder VL (2003) The immunomodulatory role of human conjunctival epithelial cells. Invest Ophthalmol Vis Sci 44:3906–3910

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. U598, INSERM, Cordeliers Biomedical Institute, 15 rue de l’école de médecine, 75006, Paris, France

    Aude Pauly, Françoise Brignole-Baudouin, Jean-Marc Guenoun, Luisa Riancho, Patrice Rat, Jean-Michel Warnet & Christophe Baudouin

  2. Department of Ophthalmology, Quinze-Vingts National Ophthalmology Hospital, 28 rue de Charenton, 75012, Paris, France

    Jean-Marc Guenoun & Christophe Baudouin

  3. Department of Toxicology, Faculty of Biological and Pharmacological Sciences, University of Paris 5 - René Descartes, Paris, France

    Aude Pauly, Françoise Brignole-Baudouin, Luisa Riancho, Patrice Rat & Jean-Michel Warnet

  4. Ambroise Paré Hospital, APHP, Paris-Ile-de-France Ouest School of Medicine, University of Versailles, Paris, France

    Christophe Baudouin

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  1. Aude Pauly
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  2. Françoise Brignole-Baudouin
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Correspondence to Christophe Baudouin.

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Commercial relationships policy: N. This study was supported by unrestricted grants from University Paris 6 and Claude Bernard Association, Paris, France.

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Pauly, A., Brignole-Baudouin, F., Guenoun, JM. et al. Comparative study of topical anti-allergic eye drops on human conjunctiva-derived cells: responses to histamine and IFNγ and toxicological profiles. Graefe's Arch Clin Exp Ophthalmol 245, 534–546 (2007). https://doi.org/10.1007/s00417-006-0353-z

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