TY - JOUR
T1 - Human RPE cell apoptosis induced by activated monocytes is mediated by caspase-3 activation
AU - Elner, Susan G.
AU - Yoshida, Ayako
AU - Bian, Zong Mei
AU - Kindezelskii, Andrei L.
AU - Petty, Howard R.
AU - Elner, Victor M.
AU - Wilson, Steven E.
AU - Klein, Ronald
AU - Mets, Marilyn B.
PY - 2003
Y1 - 2003
N2 - Purpose: To determine the effects of activated monocytes on the induction of human retinal pigment epithelial (HRPE) cell reactive oxygen metabolite (ROM) production and apoptosis. Methods: HRPE cells were co-cultured with interferon-γ (IFN-γ-stimulated human monocytes. HRPE apoptosis was detected by propidium iodide, proliferating cell nuclear antigen (PCNA) and TdT-mediated dUTP nick end labeling (TUNEL) staining, caspase-3 activation, and Western blot analysis. HRPE cell ROMs were imaged using the fluorescent marker dihydrotetramethylrosamine (H2TMRos). Results: IFN-γ-activated monocytes in direct contact with HRPE cells elicited significant increases in TUNEL-positive (P<.0001) and decreases in PCNA-positive (P<.0001) HRPE cells. The activated monocytes also induced HRPE cell caspase-3 activation, which was inhibited by inhibitor Z-DEVD-fink. Co-incubations, in which monocytes were either prevented from direct contact with HRPE cells or separated from HRPE cells after 30 minutes of direct contact, did not induce significant HRPE cell apoptosis. Anti-CD18 and anti-ICAM-1 antibodies significantly reduced activated monocyte-induced TUNEL-positive HRPE cells, by 48% (P = .0051) and 38% (P = .046), respectively, and caspase-3 activity by 56% (P<.0001) and 45% (P<.0001), respectively. Overlay of monocytes induced HRPE cell ROM that was inhibited by anti-CD18 and anti-ICAM-1 antibodies, but not by superoxide dismutase (SOD) or nitric oxide (NO) inhibitors. Accordingly, neither SOD nor NO inhibitors had significant effects on HRPE cell apoptosis or caspase-3 activation. Conclusions: We demonstrated that IFN-γ-activated monocytes may induce ROM in HRPE cells through cell-to-cell contact, in part via CD18 and ICAM-1, and promote HRPE cell apoptosis via caspase-3 activation. These mechanisms may compromise HRPE cell function and survival in retinal diseases in which mononuclear phagocyte infiltration at the HRPE interface is observed.
AB - Purpose: To determine the effects of activated monocytes on the induction of human retinal pigment epithelial (HRPE) cell reactive oxygen metabolite (ROM) production and apoptosis. Methods: HRPE cells were co-cultured with interferon-γ (IFN-γ-stimulated human monocytes. HRPE apoptosis was detected by propidium iodide, proliferating cell nuclear antigen (PCNA) and TdT-mediated dUTP nick end labeling (TUNEL) staining, caspase-3 activation, and Western blot analysis. HRPE cell ROMs were imaged using the fluorescent marker dihydrotetramethylrosamine (H2TMRos). Results: IFN-γ-activated monocytes in direct contact with HRPE cells elicited significant increases in TUNEL-positive (P<.0001) and decreases in PCNA-positive (P<.0001) HRPE cells. The activated monocytes also induced HRPE cell caspase-3 activation, which was inhibited by inhibitor Z-DEVD-fink. Co-incubations, in which monocytes were either prevented from direct contact with HRPE cells or separated from HRPE cells after 30 minutes of direct contact, did not induce significant HRPE cell apoptosis. Anti-CD18 and anti-ICAM-1 antibodies significantly reduced activated monocyte-induced TUNEL-positive HRPE cells, by 48% (P = .0051) and 38% (P = .046), respectively, and caspase-3 activity by 56% (P<.0001) and 45% (P<.0001), respectively. Overlay of monocytes induced HRPE cell ROM that was inhibited by anti-CD18 and anti-ICAM-1 antibodies, but not by superoxide dismutase (SOD) or nitric oxide (NO) inhibitors. Accordingly, neither SOD nor NO inhibitors had significant effects on HRPE cell apoptosis or caspase-3 activation. Conclusions: We demonstrated that IFN-γ-activated monocytes may induce ROM in HRPE cells through cell-to-cell contact, in part via CD18 and ICAM-1, and promote HRPE cell apoptosis via caspase-3 activation. These mechanisms may compromise HRPE cell function and survival in retinal diseases in which mononuclear phagocyte infiltration at the HRPE interface is observed.
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M3 - Article
C2 - 14971566
AN - SCOPUS:1142286550
SN - 0065-9533
VL - 101
SP - 77
EP - 91
JO - Transactions of the American Ophthalmological Society
JF - Transactions of the American Ophthalmological Society
ER -
