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H. Li, J.Y. Niederkorn, S. Neelam, J.P. McCulley, H. Alizadeh; Immunosuppressive Effect of Culture Supernatant from Human Amniotic Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):751.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Amniotic membrane has been applied to the ocular surface to restore corneal function. . However, the beneficial effect of amniotic membrane transplantation may be due to the immunosuppressive effects of amniotic epithelial cells. The aim of this study was to determine whether amniotic epithelial cells (AEC) secrete anti-inflammatory and anti-proliferative factors that affect chemotaxis of neutrophils and macrophages, and suppress both T-and B cell proliferation in vitro. Methods: Human amniotic cells were isolated from human amniotic membrane and cultured in vitro by trypsin digestion. The supernatants from AEC cultures were collected at 24, 48, and 72 hr. Neutrophil and macrophage chemotactic activity was tested in the presence of AEC supernatants using 24-well transwell chambers. Cellular proliferation was assessed using MTT and H3 thymidine incorporation. Apoptosis was examined by caspase-3 assays and expression of cytokines was assessed by RT-PCR. Results: AEC supernatants significantly inhibited chemotactic activity of neutrophils and macrophages toward macrophage inflammatory protein-2 (P< 0.05). The supernatants significantly reduced the proliferation of both T and B cells after mitogenic stimulation (P<0.05). Caspase-3 assays revealed that the supernatants induced apoptosis of T and B cells but not of corneal epithelial and stromal cells. The AEC expressed message for TNFa, Fas ligand (Fas-L), TRAIL, TRAIL 1 and 2 receptors. However, AEC induction of apoptosis was inhibited (60%) by anti-Fas-L antibody but not by anti-TRAIL antibody. Conclusions: AEC secrete soluble factors that exert anti-inflammatory effects by inhibiting chemotactic responses of neutrophils and macrophages. AEC-born factors also induce apoptosis of T and B cells by a process that relies on Fas-Fas-L interactions.
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