Top

Graefe's Archive for Clinical and Experimental Ophthalmology

Published in:

01-11-2014 | Basic Science

The retinal pigment epithelium (RPE) induces FasL and reduces iNOS and Cox2 in primary monocytes

Authors: Christin Hettich, Sebastian Wilker, Rolf Mentlein, Ralph Lucius, Johann Roider, Alexa Klettner

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 11/2014

Abstract

Purpose

Retinal pigment epithelium (RPE) cells may alter the phenotype of monocytes by soluble factors that may be influenced by stimulation of the RPE. Since RPE cells carry the toll-like receptor-3 (TLR3) that detects and reacts to viral infection through binding of dsRNA we investigated the effects of RPE cells with or without TLR3 stimulation on blood-derived monocytes with respect to regulation of pro-/anti-inflammatory cytokines, anti-angiogenic factors and migratory properties.

Methods

Primary RPE cells were prepared from porcine eyes; monocytes were prepared from porcine blood. TLR3 activation was induced by polyinosinic:polycytidylic acid (Poly I:C). RPE cells were stimulated with Poly I:C in different concentrations for 24 hours and a cell culture supernatant was applied to the monocytes. Expression of CD14 and Fas ligand (FasL) was determined via flow cytometry. The expression of IL-6, IL-1ß, TNFα, Cox2, iNOS and IL-10 was determined via quantitative RT-PCR. Migration was determined using Boyden chamber experiments.

Results

The supernatant of RPE cells, irrespective of TLR3 activation, induced FasL expression in the monocytes. Expression of iNOS and Cox2 was reduced by RPE cells and the reduction of Cox2 but not if iNOS was lost under TLR3 activation. No induction of IL-6, IL-1ß, IL-10 or TNFα by the RPE was seen. TLR3-activated RPE cells induced monocyte migration.

Conclusion

RPE cells induce an upregulation of FasL and a downregulation of iNOS and Cox2 without upregulating inflammatory cytokines, possibly inducing an anti-angiogenic phenotype in the monocytes. This phenotype is still upheld after challenging RPE cells with dsRNA, mimicking a viral infection.

Espinosa-Heidmann DG, Suner IJ, Hernandez EP, Monroy D, Csaky KG, Cousins SW (2003) Macrophage depletion diminishes lesion size and severity in experimental choroidal neovascularization. Invest Ophthalmol Vis Sci 44:3586–3592PubMedCrossRef

Apte RS, Richter J, Herndon J, Ferguson TA (2006) Macrophages inhibit neovascularization in a murine model of age-related macular degeneration. PLoS Med 3:e310

Ferguson TA, Griffith TS (2006) A vision of cell death: Fas ligand and immune privilege 10 years later. Immunol Rev 213:228–238PubMedCrossRef

Kelly J, Ali Khan A, Yin J, Ferguson TA, Apte RS (2007) Senescence regulates macrophage activation and angiogenic fate at sites of tissue injury in mice. J Clin Invest 117:3421–3426PubMedCentralPubMedCrossRef

Strauss O (2005) The retinal pigment epithelium in visual function. Physiol Rev 85:845–881PubMedCrossRef

Detrick B, Hooks JJ (2010) Immune regulation in the retina. Immunol Res 47:153–161PubMedCrossRef

Zamiri P, Masli S, Kitaichi N, Taylor AW, Streilein JW (2005) Thrombospondin plays a vital role in the immune privilege of the eye. Invest Ophthalmol Vis Sci 46:908–919PubMedCrossRef

Sugita S, Futagami Y, Smith SB, Naggar H, Mochizuki M (2006) Retinal and ciliary body pigment epithelium suppress activation of T lymphocytes via transforming growth factor beta. Exp Eye Res 83:1459–1471PubMedCrossRef

Vega JL, Saban D, Carrier Y, Masli S, Weiner HL (2010) Retinal pigment epithelial cells induce foxp3(+) regulatory T cells via membrane-bound TGF-ß. Ocul Immunol Inflamm 18:459–469PubMedCrossRef

10.

Gregerson DS, Heuss ND, Lew KL, McPherson SW, Ferrington DA (2007) Interaction of retinal pigmented epithelial cells and CD4 T cells leads to T-cellanergy. Invest Ophthalmol Vis Sci 48:4654–4663

11.

Ferguson TA, Apte RS (2008) Angiogenesis in eye disease: immunity gained or immunity lost? Semin Immunopathol 30:111–119PubMedCrossRef

12.

Jørgensen A, Wiencke AK, la Cour M, Kaestel CG, Madsen HO, Hamann S, Lui GM, Scherfig E, Prause JU, Svejgaard A, Odum N, Nissen MH, Röpke C (1998) Human retinal pigment epithelial cell-induced apoptosis in activated T cells. Invest Ophthalmol Vis Sci 39:1590–1599PubMed

13.

Huemer HP, Larcher C, Kirchebner W, Klingenschmid J, Göttinger W, Irschick EU (1996) Susceptibility of human retinal epithelial cells to different viruses. Graefes Arch Clin Exp Ophthalmol 234:177–185PubMedCrossRef

14.

Kaarniranta K, Salminen A (2009) Age-relatedmacular degeneration: activation of innate immunity system via pattern recognition receptors. J Mol Med 87:117–123

15.

Bian ZM, Elner SG, Yoshida A, Elner VM (2003) Human RPE-monocyte co-culture induces chemokine gene expression through activation of MAPK and NIK cascade. Exp Eye Res 76:573–583PubMedCrossRef

16.

Elner VM, Strieter RM, Elner SG, Baggiolini M, Lindley I, Kunkel SL (1990) Neutrophil chemotactic factor (IL-8) gene expression by cytokine-treated retinal pigment epithelial cells. Am J Pathol 136:745–750PubMedCentralPubMed

17.

Elner SG, Yoshida A, Bian ZM, Kindezelskii AL, Petty HR, Elner VM (2003) Human RPE cell apoptosis induced by activated monocytes is mediated by caspase-3 activation. Trans Am Ophthalmol Soc 101:77–92PubMedCentralPubMed

18.

Cherepanoff S, McMenamin P, Gillies MC, Kettle E, Sarks SH (2010) Bruch’s membrane and choroidal macrophages in early and advanced age-related macular degeneration. Br J Ophthalmol 94:918–925PubMedCrossRef

19.

Jerdan JA, Pepose JS, Michels RG, Hayashi H, de Bustros S, Sebag M, Glaser BM (1989) Proliferative vitreoretinopathy membranes. An immunohistochemical study. Ophthalmology 96:801–810PubMedCrossRef

20.

Yang P, de Vos AF, Kijlstra A (1997) Macrophages and MHC class II positive cells in the choroid during endotoxin induced uveitis. Br J Ophthalmol 81:396–401PubMedCentralPubMedCrossRef

21.

Lau CH, Taylor AW (2009) The immune privileged retina mediates an alternative activation of J774A.1 cells. Ocul Immunol Inflamm 17:380–389PubMedCrossRef

22.

Matsumoto M, Kikkawa S, Kohase M, Miyake K, Seya T (2002) Establishment of a monoclonal antibody against human Toll-like receptor 3 that blocks double-stranded RNA-mediated signalling. Biochem Biophys Res Commun 293:1364–1369

23.

Doyle SE, O’Connell R, Vaidya SA, Chow EK, Yee K, Cheng G (2003) Toll-likereceptor 3 mediates a more potent antiviral response that Toll-likereceptor 4. J Immunol 170:3565–3571

24.

Kleinman ME, Kaneko H, Cho WG, Dridi S, Fowler BJ, Blandford AD, Albuquerque RJ, Hirano Y, Terasaki H, Kondo M, Fujita T, Ambati BK, Tarallo V, Gelfand BD, Bogdanovich S, Baffi JZ, Ambati J (2012) Short-interfering RNAs induce retinal degeneration via TLR3 and IRF3. Mol Ther 20:101–108

25.

McLeod DS, Grebe R, Bhutto I, Merges C, Baba T, Lutty GA (2009) Relationship between RPE and choriocapillaris in age-related macular degeneration. Invest Ophthalmol Vis Sci 50:4982–4991PubMedCrossRef

26.

Yang Z, Stratton C, Francis PJ, Kleinman ME, Tan PL, Gibbs D, Tong Z, Chen H, Constantine R, Yang X, Zeng J, Davey L, Ma X, Hau VS, Wang C, Harmon J, Buehler J, Pearson E, Patel S, Kaminoh Y, Watkins S, Luo L, Zabriskie NA, Bernstein PS, Cho W, Schwager A, Hinton DR, Klein ML, Hamon SC, Simmons E, Yu B, Campochiaro B, Sunness JS, Campochiaro P, Jorde L, Parmigiani G, Zack DJ, Katsanis N, Ambati J, Zhang K (2008) Toll-likereceptor 3 and geographic atrophy in age-related macular degeneration. N Engl J Med 359:1456–1463

27.

Cho Y, Wang JJ, Chew E, Ferris FL, Mitchell P, Chan CC, Tuo J (2009) Toll-like receptor polymorphisms and age-related macular degeneration: replication in three case–control samples. Invest Ophthalmol Vis Sci 50:5614–5618PubMedCentralPubMedCrossRef

28.

Zhou P, Fan L, Yu KD, Zhao MW, Li XX (2011) Toll-likereceptor 3 C1234T may protect against geographic atrophy through decreased dsRNA binding capacity. FASEB J 25:3489–3495

29.

Kumar MV, Nagineni CN, Chin MS, Hooks JJ, Detrick B (2004) Innate immunity in the retina: Toll-like receptor (TLR) signaling in human retina pigment epithelial cells. J Neuroimmunol 153:7–15PubMedCrossRef

30.

Ebihara N, Chen L, Tokura T, Ushio H, Iwatsu M, Murakami A (2007) Distinct functions between toll-like receptors 3 and 9 in retinal pigment epithelial cells. Ophthalmic Res 39:155–163PubMedCrossRef

31.

Klettner A, Koinzer S, Meyer T, Roider J (2013) Toll-like receptor 3 activation in retinal pigment epithelium cells – Mitogen-activated protein kinase pathways of cell death and vascular endothelial growth factor secretion. Acta Ophthalmol 91:e211–e218

32.

Klettner A, Roider J (2008) Comparison of bevacizumab, ranibizumab, and pegaptanib: efficiency and possible additional pathways. Invest Ophthalmol Vis Sci 49:4523–4527PubMedCrossRef

33.

Wiencke AK, Kiilgaard JF, Nicolini J, Bundgaard M, Röpke C, la Cour M (2003) Growth of cultured porcine retinal pigment epithelial cells. Acta Ophthalmol 81:170–176CrossRef

34.

Berg C, Wilker S, Roider J, Klettner A (2013) Isolation of porcine monocyte population: a simple and efficient method. Vet Res Commun 37:239–241PubMedCrossRef

35.

Ziegler-Heitbrock HW, Appl B, Käfferlein E, Löffler T, Jahn-Henninger H, Gutensohn W, Nores JR, Mccullough K, Passlick B, Labeta MO et al (1994) The Antibody MY4 Recognizes CD14 on Porcine Monocytes and Macrophages. Scand J Immunol 40:509–514PubMedCrossRef

36.

Klettner A, Baumgrass R, Zhang Y, Fischer G, Bürger E, Herdegen T, Mielke K (2001) The neuroprotective actions of FK506 binding protein ligands: neuronal survival is triggered by de novo RNA synthesis, but is independent of inhibition of JNK and Calcineurin. Brain Res Mol Brain Res 97:21–31PubMedCrossRef

37.

Tang S, Lucius R, Wenck H, Gallinat S, Weise JM (2013) UV-mediated downregulation of the endocytic collagen receptor, Endo 180, contributes to accumulation of extracellular collagen fragments in photoaged skin. J Dermatol Sci 70:42–48PubMedCrossRef

38.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR an the 2(−Delta Delta C (T)) method. Methods 25:402–408

39.

Liles WC, Kiener PA, Ledbetter JA, Aruffo A, Klebanoff SJ (1996) Differential expression of Fas (CD95) and Fas ligand on normal human phagocytes: implications for the regulation of apoptosis in neutrophils. J Exp Med 184:429–440PubMedCrossRef

40.

Brown SB, Savill J (1999) Phagocytosis triggers macrophage release of Fas ligand and induces apoptosis of bystander leukocytes. J Immunol 162:480–485PubMed

41.

Kaplan HJ, Leibole MA, Tezel T, Ferguson TA (1999) Fas ligand (CD95 ligand) controls angiogenesis beneath the retina. Nat Med 5:292–297PubMedCrossRef

42.

Davis MH, Eubanks JP, Powers MR (2003) Increased retinal neovascularization in Fas ligand-deficient mice. Invest Ophthalmol Vis Sci 44:3202–3210CrossRef

43.

Griffith TS, Brunner T, Fletcher SM, Green DR, Ferguson TA (1995) Fas ligand-induced apoptosis as a mechanism of immune privilege. Science 270:1189–1192PubMedCrossRef

44.

Yoshida A, Elner SG, Bian ZM, Kindezelskii AL, Petty HR, Elner VM (2003) Activated monocytes induce human retinal pigment epithelial cell apoptosis through caspase-3 activation. Lab Investig 83:1117–1129PubMedCrossRef

45.

Esser P, Heimann K, Abts H, Fontana A, Weller M (1995) CD95 (Fas/APO-1) antibody-mediated apoptosis of human retinal pigment epithelial cells. Biochem Biophys Res Commun 213:1026–1034PubMedCrossRef

46.

Rosenbaum JT, O’Rourke L, Davies G, Wenger C, David L, Robertson JE (1987) Retinal pigment epithelial cells secrete substances that are chemotactic for monocytes. Curr Eye Res 6:793–800PubMedCrossRef

47.

Yoshida A, Elner SG, Bian ZM, Kunkel SL, Lukacs NW, Elner VM (2001) Thrombin regulates chemokine induction during human retinal pigment epithelial cell/monocyte interaction. Am J Pathol 159:1171–1180PubMedCentralPubMedCrossRef

48.

Yang D, Elner SG, Chen X, Field MG, Petty HR, Elner VM (2011) MCP-1 activated monocytes induce apoptosis in human retinal pigment epithelium. Invest Ophthalmol Vis Sci 52:6026–6034

49.

Ando A, Yang A, Nambu H, Campochiaro PA (2002) Blockade of nitric-oxide synthase reduces choroidal neovascularization. Mol Pharmacol 62:539–544PubMedCrossRef

50.

Ando A, Yang A, Mori K, Yamada H, Yamada E, Takahashi K, Saikia J, Kim M, Melia M, Fishman M, Huang P, Campochiaro PA (2002) Nitric oxide is proangiogenic in the retina and choroid. J Cell Physiol 191:116–124PubMedCrossRef

51.

Houssier M, Raoul W, Lavalette S, Keller N, Guillonneau X, Baragatti B, Jonet L, Jeanny JC, Behar-Cohen F, Coceani F, Scherman D, Lachapelle P, Ong H, Chemtob S, Sennlaub F (2008) CD36 deficiency leads to choroidal involution via COX2 down-regulation in rodents. PLoS Med 5:e39

52.

Klettner A, Hamann T, Schlüter K, Lucius R, Roider J (2014) Retinal pigment epithelium cells alter the pro-inflammatory response of retinal microglia to TLR-3 stimulation. Acta Ophthalmol. doi:10.1111/aos.12472

53.

Lassota N (2008) Clinical and histological aspects of CNV formation: studies in an animal model. Acta Ophthalmol 86:1–24PubMedCrossRef

54.

Sanchez I, Martin R, Ussa F, Fernandez-Bueno I (2011) The parameters of the porcine eyeball. Graefes Arch Clin Exp Ophthalmol 249:475–482

55.

Middleton S (2010) Porcine Ophthalmology. Vet Clin N Am Food Anim Pract 26:557–572CrossRef

56.

Butler JE, Sun J, Wertz N, Sinkora M (2006) Antibody repertoire development in swine. Dev Comp Immunol 30:199–211PubMedCrossRef

57.

Yang P, Chen L, Zwart R, Kijlstra A (2002) Immune cells in the porcine retina: Distribution, characterization and morphological features. Invest Ophthalmol Vis Sci 43:1488–1492PubMed

Title: The retinal pigment epithelium (RPE) induces FasL and reduces iNOS and Cox2 in primary monocytes
Authors: Christin Hettich
Sebastian Wilker
Rolf Mentlein
Ralph Lucius
Johann Roider
Alexa Klettner
Publication date: 01-11-2014
Publisher: Springer Berlin Heidelberg
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 11/2014
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-014-2742-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

The retinal pigment epithelium (RPE) induces FasL and reduces iNOS and Cox2 in primary monocytes

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 11/2014

Retinal vessel diameter in normal-tension glaucoma patients with asymmetric progression

An analysis of the outcomes for patients with failed primary vitrectomy for rhegmatogenous retinal detachment

Comparison of deep anterior lamellar keratoplasty and penetrating keratoplasty with respect to postoperative corneal sensitivity and tear film function

High-frequency electric welding: a novel method for improved immediate chorioretinal adhesion in vitreoretinal surgery

Posterior segment findings of ocular cytomegalovirus infection in immunocompetent patients

Early axonal damage detection by ganglion cell complex analysis with optical coherence tomography in nonarteritic anterior ischaemic optic neuropathy