| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 From the Department of Biochemistry and 2 Department of Ophthalmology, Emory University School of Medicine, Atlanta, Georgia.
PURPOSE. To investigate whether Fas ligand (FasL) and the Fas receptor system mediates apoptosis in cultured human retinal pigment epithelial (hRPE) cells and contributes to oxidant-induced death of hRPE cells.
METHODS. Expression of FasL and Fas in cultured hRPE cells was examined by Western blot analysis and flow cytometry. The susceptibility of hRPE cells to Fas-mediated apoptosis was determined by incubating cells with recombinant soluble Fas ligand (sFasL). Characteristics of apoptosis assessed included chromatin condensation, DNA cleavage, and phosphatidylserine exposure. To investigate the possible involvement of Fas-mediated apoptosis in oxidative killing of hRPE cells, the effects of the oxidant tert-butylhydroperoxide (tBH) on the expression of FasL and Fas were studied. The specificity of effects of oxidant was examined using the antioxidants glutathione and N-acetyl-L-cysteine (NAC). The requirement for the Fas pathway in tBH-induced apoptosis was investigated using an antagonistic anti-Fas antibody ZB4 that blocks the interaction between FasL and Fas.
RESULTS. Cultured hRPE cells constitutively expressed FasL and Fas. Ligation of Fas receptor with recombinant sFasL triggered apoptosis in hRPE cells. tBH treatment of hRPE cells resulted in increased expression of FasL and Fas. Glutathione and NAC completely abrogated tBH-induced increase in FasL and Fas expression and apoptosis. Blocking FasL and Fas interaction by ZB4 inhibited tBH-induced apoptosis, but only partially.
CONCLUSIONS. A functional Fas-mediated apoptotic pathway is present in cultured hRPE cells and can be activated with sFasL or by upregulation of FasL and Fas expression with an oxidant. The incomplete inhibition by blocking antibody indicates that the Fas pathway is involved in oxidant-induced apoptosis, but other triggering mechanisms are also important.
This article has been cited by other articles:
|
|
 |
|
  S. Jiang, S. E. Moriarty-Craige, C. Li, M. J. Lynn, J. Cai, D. P. Jones, and P. Sternberg Associations of Plasma-Soluble Fas Ligand with Aging and Age-Related Macular Degeneration Invest. Ophthalmol. Vis. Sci., April 1, 2008; 49(4): 1345 - 1349. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Chen, P. Sternberg, and J. Cai Characterization of a Bcl-XL-Interacting Protein FKBP8 and Its Splice Variant in Human RPE Cells Invest. Ophthalmol. Vis. Sci., April 1, 2008; 49(4): 1721 - 1727. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Neekhra, S. Luthra, M. Chwa, G. Seigel, A. L. Gramajo, B. D. Kuppermann, and M. C. Kenney Caspase-8, -12, and -3 Activation by 7-Ketocholesterol in Retinal Neurosensory Cells Invest. Ophthalmol. Vis. Sci., March 1, 2007; 48(3): 1362 - 1367. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. S. Gregerson, K. L. Lew, S. W. McPherson, N. D. Heuss, and D. A. Ferrington RPE Cells Resist Bystander Killing by CTLs, but Are Highly Susceptible to Antigen-Dependent CTL Killing Invest. Ophthalmol. Vis. Sci., December 1, 2006; 47(12): 5385 - 5394. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Luthra, B. Fardin, J. Dong, D. Hertzog, S. Kamjoo, S. Gebremariam, V. Butani, R. Narayanan, J. K. Mungcal, B. D. Kuppermann, et al. Activation of Caspase-8 and Caspase-12 Pathways by 7-Ketocholesterol in Human Retinal Pigment Epithelial Cells Invest. Ophthalmol. Vis. Sci., December 1, 2006; 47(12): 5569 - 5575. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-N. Ha, Y. Chen, J. Cai, and P. Sternberg Jr Increased Glutathione Synthesis through an ARE-Nrf2-Dependent Pathway by Zinc in the RPE: Implication for Protection against Oxidative Stress. Invest. Ophthalmol. Vis. Sci., June 1, 2006; 47(6): 2709 - 2715. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Lincoln, M. Boling, A. N. Parikh, Y. Yeh, D. G. Gilchrist, and L. S. Morse Fas Signaling Induces Raft Coalescence That Is Blocked by Cholesterol Depletion in Human RPE Cells Undergoing Apoptosis Invest. Ophthalmol. Vis. Sci., May 1, 2006; 47(5): 2172 - 2178. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Jiang, S. E. Moriarty-Craige, M. Orr, J. Cai, P. Sternberg Jr, and D. P. Jones Oxidant-Induced Apoptosis in Human Retinal Pigment Epithelial Cells: Dependence on Extracellular Redox State Invest. Ophthalmol. Vis. Sci., March 1, 2005; 46(3): 1054 - 1061. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. L. Dunaief, T. Dentchev, G.-S. Ying, and A. H. Milam The Role of Apoptosis in Age-Related Macular Degeneration Arch Ophthalmol, November 1, 2002; 120(11): 1435 - 1442. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Jiang, S. E. Moriarty, H. Grossniklaus, K. C. Nelson, D. P. Jones, and P. Sternberg Jr Increased Oxidant-Induced Apoptosis in Cultured Nondividing Human Retinal Pigment Epithelial Cells Invest. Ophthalmol. Vis. Sci., August 1, 2002; 43(8): 2546 - 2553. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Strunnikova, J. Baffi, A. Gonzalez, W. Silk, S. W. Cousins, and K. G. Csaky Regulated Heat Shock Protein 27 Expression in Human Retinal Pigment Epithelium Invest. Ophthalmol. Vis. Sci., August 1, 2001; 42(9): 2130 - 2138. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W.-Y. Shen, M. C. Lai, J. Beilby, N. L. Barnett, J. Liu, I. J. Constable, and P. E. Rakoczy Combined Effect of Cyclosporine and Sirolimus on Improving the Longevity of Recombinant Adenovirus-Mediated Transgene Expression in the Retina Arch Ophthalmol, July 1, 2001; 119(7): 1033 - 1043. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A. Chrestensen, D. W. Starke, and J. J. Mieyal Acute Cadmium Exposure Inactivates Thioltransferase (Glutaredoxin), Inhibits Intracellular Reduction of Protein-glutathionyl-mixed Disulfides, and Initiates Apoptosis J. Biol. Chem., August 18, 2000; 275(34): 26556 - 26565. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
