Skip to main content
SpringerLink
Log in

Transforming growth factor-β regulates human retinal pigment epithelial cell phagocytosis by influencing a protein kinase C-dependent pathway

  • Laboratory Investigation
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

• Background: Transforming growth factor-β (TGF-β) plays an important role in the pathogenesis of many ocular diseases, including proliferative vitreoretinopathy. We examined the effect of TGF-β on the phagocytosis of rod outer segments by retinal pigment epithelium (RPE), which is a major function of RPE, and investigated the dependence of this effect on the protein kinase C (PKC) pathway. • Methods: Phagocytotic uptake of fluoresceinated bovine rod outer segments was determined by flow cytometry. RPE cells were treated with TGF-β1 or TGF-β2 and their effects on phagocytosis were examined. The effects of various PKC inhibitors (calphostin C, staurosporine, and extended exposure to phorbol 12-myristate 13-acetate, PMA) and a stimulator (brief exposure to PMA) on RPE phagocytosis was evaluated. • Results: Both TGF-β1 and TGF-β2 up-regulated RPE phagocytosis and PMA abolished the upregulating effect of TGF-β. In contrast, PKC inhibition by staurosporine and calphostin C resulted in increased phagocytosis. A combination of TGF-β and PKC inhibitor treatment did not produced any additive effect on phagocytosis. • Conclusion: We concluded that TGF-β up-regulates human RPE phagocytosis, but that this effect is counteracted by PKC activation. It is possible that this TGF-β-induced effect is due, in part, to a negative modulation of the PKC-dependent pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Anderson DH, Stern WH, Fisher SK, Erickson PA, Borgula GA (1983) Retinal detachment in the cat: the pigment epithelial-photoreceptor interface. Invest Ophthalmol Vis Sci 24:906–926

    Google Scholar 

  2. Barrett TB, Shu GL, Draves KE, Pezzutto A, Clark EA (1990) Signaling through CD19, Fc receptors or transforming growth factor-β: each inhibits the activation of resting human B cells differently. Eur J Immunol 20:1053–1059

    Google Scholar 

  3. Belosevic M, Daniels CW (1992) Phagocytosis of Giardia lamblia trophozoites by cytokine-activated macrophages. Clin Exp Immunol 87:304–309

    Google Scholar 

  4. Boyle DL, McLaughlin BJ (1990) The effect of swainsonine on the phagocytosis of rod outer segments by rat RPE. Curr Eye Res 9:407–414

    Google Scholar 

  5. Boyle D, Tien L, Cooper NGF, Shepherd V, McLaughlin BJ (1991) A mannose receptor is involved in retinal phagocytosis. Invest Ophthalmol Vis Sci 32:1464–1470

    Google Scholar 

  6. Cantrell DA, Davies AA, Crumpton MJ (1985) Activators of protein kinase C down-regulate and phosphorylate the T3/T-cell antigen receptor complex of human T lymphocytes. Proc Natl Acad Sci USA 82:8158–8162

    Google Scholar 

  7. Cleary PE, Ryan SJ (1979) Experimental posterior penetrating eye injury in the rabbit. II. Histology of wound, vitreous, and retina. Br J Ophthalmol 63:312–321

    Google Scholar 

  8. Connor TB Jr, Roberts AB, Sporn MB, Danielpour D, Dart LL, Michels RG, de Bustros S, Enger C, Kato H, Lansing M, Hayashi H, Glaser BM (1989) Correlation of fibrosis and transforming growth factor-β type 2 levels in the eye. J Clin Invest 83:1661–1666

    Google Scholar 

  9. Cousins SW, McCabe MM, Danielpour D, Streilein JW (1991) Identification of transforming growth factor-beta as an immunosuppressive factor in aqueous humor. Invest Ophthalmol Vis Sci 32:2201–2211

    Google Scholar 

  10. Del Monte MA, Maumenee IH (1980) New technique for in vitro culture of human retinal pigment epithelium. Birth Defects 16(2):327–338

    Google Scholar 

  11. Eisenstein R, Grant-Bertacchini D (1991) Growth inhibitory activities in avascular tissues are recognized by anti-transforming growth factor β antibodies. Curr Eye Res 10:157–162

    Google Scholar 

  12. Gaudric A, Glacet-Bernard A, Clément G, Falquerho L, Barritault D, Coscas G (1990) Transforming growth factor bêta dans le vitré de patients présentant une proliferation épirétinienne. Ophtalmologie 4:51–52

    Google Scholar 

  13. Gopalakrishna R, Chen ZH, Gundimeda U (1992) Irreversible oxidative inactivation of protein kinase C by photosensitive inhibitor calphostin C. FEBS Lett 314:149–154

    Google Scholar 

  14. Gresham HD, Zheleznyak A, Mormol JS, Brown EJ (1990) Studies on the molecular mechanisms of human neutrophil Fc receptor-mediated phagocytosis. Evidence that a distinct pathway for activation of the respiratory burst results in reactive oxygen metabolite-dependent amplification of ingestion. J Biol Chem 265:7819–7826

    Google Scholar 

  15. Gresham HD, Ray CJ, O'Sullivan FX (1991) Defective neutrophil function in the autoimmune mouse strain MRL/1pr: potential role of transforming growth factor-β. J Immunol 146:3911–3921

    Google Scholar 

  16. Hall MO, Abrams T (1987) Kinetic studies of rod outer segment binding and ingestion by cultured rat RPE cells. Exp Eye Res 45:907–922

    Google Scholar 

  17. Hall MO, Burgess BL, Arakawa H, Fliesler SJ (1990) The effect of inhibitors of glycoprotein synthesis and processing on the phagocytosis of rod outer segments by cultured retinal pigment epithelial cells. Glycobiology 1:51–61

    Google Scholar 

  18. Hall MO, Abrams TA, Mittag TW (1991) ROS ingestion by RPE cells is turned off by increased protein kinase C activity and by increased calcium. Exp Eye Res 52:591–598

    Google Scholar 

  19. Hishikawa T, Cheung JY, Yelamarty RV, Knutson DW (1991) Calcium transients during Fc receptor-mediated and nonspecific phagocytosis by murine peritoneal macrophages. J Cell Biol 115:59–66

    Google Scholar 

  20. Howe PH, Bascom CC, Cunningham MR, Leof EB (1989) Regulation of transforming growth factor β1 action by multiple transducing pathways: evidence for both G protein-dependent and -independent signaling. Cancer Res 49:6024–6031

    Google Scholar 

  21. Ito M, Yasui W, Kyo E, Yokozaki H, Nakayama H, Ito H, Tahara E (1992) Growth inhibition of transforming growth factor β on human gastric carcinoma cells: receptor and postreceptor signaling. Cancer Res 52:295–300

    Google Scholar 

  22. Kamijo R, Takeda K, Nagumo M, Konno K (1990) Effects of combinations of transforming growth factor-β1 and tumor necrosis factor on induction of differentiation of human myelogenous leukemic cell lines. J Immunol 144:1311–1316

    Google Scholar 

  23. Kikkawa U, Kishimoto A, Nishizuka Y (1989) The protein kinase C family: heterogeneity and its implications. Annu Rev Biochem 58:31–44

    Google Scholar 

  24. Knowles GC, McKeown M, Sodek J, McCulloch CAG (1991) Mechanism of collagen phagocytosis by human gingival fibroblasts: importance of collagen structure in cell recognition and internalization. J Cell Sci 98:551–558

    Google Scholar 

  25. Kobayashi E, Nakano H, Morimoto M, Tamaoki T (1989) Calphostin C (UCN-1028C), a novel microbial compound, is a highly potent and specific inhibitor of protein kinase C. Biochem Biophys Res Commun 159:548–553

    Google Scholar 

  26. Laird DW, Molday RS (1988) Evidence against the role of rhodopsin in rod outer segment binding to RPE cells. Invest Ophthalmol Vis Sci 29:419–428

    Google Scholar 

  27. Lutty GA, Merges C, Threlkeld AB, Crone S, McLeod DS (1993) Heterogeneity in localization of isoforms of TGF-β in human retina, vitreous, and choroid. Invest Ophthalmol Vis Sci 34:477–487

    Google Scholar 

  28. Machemer R, Laqua H (1975) Pigment epithelium proliferation in retinal detachment (massive periretinal proliferation). Am J Ophthalmol 80:1–23

    CAS  PubMed  Google Scholar 

  29. Mano T, Puro DG (1990) Phagocytosis by human retinal glial cells in culture. Invest Ophthalmol Vis Sci 31:1047–1055

    Google Scholar 

  30. Markovac J, Goldstein GW (1988) Transforming growth factor beta activates protein kinase C in microvessels isolated from immature rat brain. Biochem Biophys Res Commun 150:575–582

    Google Scholar 

  31. Michishita M, Hirayoshi K, Tsuru A, Nakamura N, Yoshida Y, Okuma M, Nagata K (1991) Effects of type-β1 transforming growth factor on the proliferation and differentiation of mouse myelomonocytic leukemia cells (M1). Exp Cell Res 196:107–113

    Google Scholar 

  32. Muldoon LL, Rodland KD, Magun BE (1988) Transforming growth factor β and epidermal growth factor alter calcium influx and phosphatidylinositol turnover in Rat-1 fibroblasts. J Biol Chem 263:18834–18841

    Google Scholar 

  33. Newman SL, Mikus LK, Tucci MA (1991) Differential requirements for cellular cytoskeleton in human macrophage complement receptorand Fc receptor-mediated phagocytosis. J Immunol 146:967–974

    Google Scholar 

  34. Okabe-Kado J, Honma Y, Hayashi M, Hozumi M (1989) Inhibitory action of transforming growth factor-β on induction of differentiation of myeloid leukemia cells. Jpn J Cancer Res 80:228–232

    Google Scholar 

  35. Papermaster DS (1982) Preparation of retinal rod outer segments. Methods Enzymol 81:48–52

    Google Scholar 

  36. Park CH, Latina MA (1993) Effects of gamma-interferon on human trabecular meshwork cell phagocytosis. Invest Ophthalmol Vis Sci 34:2228–2236

    Google Scholar 

  37. Pasquale LR, Dorman-Pease ME, Lutty GA, Quigley HA, Jampel HD (1993) Immunolocalization of TGF-β1, TGF-β2, and TGF-β3 in the anterior segment of the human eye. Invest Ophthalmol Vis Sci 34:23–30

    Google Scholar 

  38. Raymond MC, Thompson JT (1990) RPE-mediated collagen gel contraction: inhibition by colchicine and stimulation by TGF-beta. Invest Ophthalmol Vis Sci 31:1079–1086

    Google Scholar 

  39. Robertson PL, Markovac J, Datta SC, Goldstein GW (1988) Transforming growth factor beta stimulates phosphoinositol metabolism and translocation of protein kinase C in cultured astrocytes. Neurosci Lett 93:107–113

    Google Scholar 

  40. Roubey RAS, Ross GD, Merrill JT, Walton F, Reed W, Winchester RJ, Buyon JP (1991) Staurosporine inhibits neurotrophil phagocytosis but not iC3b binding mediated by CR3 (CD11b/CD18). J Immunol 146:3557–3562

    Google Scholar 

  41. Schlessinger J (1986) Allosteric regulation of the epidermal growth factor receptor kinase. J Cell Biol 103:2067–2072

    Google Scholar 

  42. Shepherd VL, Tarnowski BI, McLaughlin BJ (1991) Isolation and characterization of a mannose receptor from human pigment epithelium. Invest Ophthalmol Vis Sci 32:1779–1784

    Google 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 135:397–402

    Google Scholar 

  44. Tanihara H, Yoshida M, Matsumoto M, Yoshimura N (1993) Identification of transforming growth factor-β expressed in cultured human retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 34:413–419

    Google Scholar 

  45. Tsunawaki S, Sporn M, Ding A, Nathan C (1988) Deactivation of macrophages by transforming growth factor-β. Nature 334:260–262

    Google Scholar 

  46. Weller M, Heimann K, Wiedemann P (1990) Mononukläre Phagozyten und ihre Wachstumsfaktoren: Schrittmacher der proliferativen Vitreoretinopathie. Klin Monatsbl Augenheilkd 196:121–127

    Google Scholar 

  47. Williams LL, Lew HM, Shannon BT, Singley CT, Davidorf FH, Jin R, Wolinsky JS (1993) Phagocytosis of latex beads is defective in cultured human retinal pigment epithelial cells with persistent rubella virus infection. Am J Pathol 142:451–461

    Google Scholar 

  48. Zheleznyak A, Brown EJ (1992) Immunoglobulin-mediated phagocytosis by human monocytes requires protein kinase C activation: evidence for protein kinase C translocation to phagosomes. J Biol Chem 267:12042–12048

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Ophthalmology, University of Southern California, School of Medicine, 1333 San Pablo Street, 90033, Los Angeles, CA, USA

    Shwu-Jiuan Sheu, Taiji Sakamoto, Roman Osusky, Hsin-Min Wang, Thomas E. Ogden & Stephen J. Ryan

  2. Doheny Eye Institute, 1450 San Pablo Street, 90033-4681, Los Angeles, CA, USA

    Shwu-Jiuan Sheu, Taiji Sakamoto, Roman Osusky, Hsin-Min Wang, Thomas E. Ogden & Stephen J. Ryan

  3. Department of Pathology, University of Southern California, School of Medicine, 1333 San Pablo Street, 90033, Los Angeles, CA, USA

    David R. Hinton

  4. Department of Pharmacology, University of Southern California, School of Medicine, Zonal Avenue, 90033, Los Angeles, CA, USA

    Rayudu Gopalakrishna

Authors
  1. Shwu-Jiuan Sheu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Taiji Sakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Roman Osusky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hsin-Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Thomas E. Ogden
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Stephen J. Ryan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. David R. Hinton
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Rayudu Gopalakrishna
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sheu, SJ., Sakamoto, T., Osusky, R. et al. Transforming growth factor-β regulates human retinal pigment epithelial cell phagocytosis by influencing a protein kinase C-dependent pathway. Graefe's Arch Clin Exp Ophthalmol 232, 695–701 (1994). https://doi.org/10.1007/BF00171387

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00171387

Keywords

Search

Navigation