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D. Bauer, M. Hennig, S. Wasmuth, M. Busch, H. Baehler, K.-P. Steuhl, A. Heiligenhaus; Phagocytosis of Apoptotic or Necrotic Cells Facilitate Macrophage Survival and Proliferation in Necrotizing Hsv-1 Keratitis in the Presence of Amniotic Membrane. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1947.
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© ARVO (1962-2015); The Authors (2016-present)
With amniotic membrane transplantation (AMT) a rapid induction of lymphocyte and PMN apoptosis and their removal by macrophages is noted in corneas with ulcerative herpetic stromal keratitis (HSK) in mice. Herein we determined the content of macrophages in HSK corneas after AMT. We furthermore determined the influence of AM in vitro on proliferation and viability of bone marrow derived macrophages (BM).
BALB/c mice were corneally infected with 1x105 PFU HSV-1. On day 14, mice with ulcerative HSK were treated by AMT or tarsorrhapy (T). Corneal cells were stained with antibodies targeting CD45 (FITC), Annexin-V (PE), 7-AAD, F4/80 (Alexa-Fluor 647) after 12 h and analysed by flow cytometry. BM cells were co-cultured with AM together with apoptotic or necrotic cell bodies. The proliferative response of BM to IFN-gamma or LPS was investigated by uptake of 3H-thymidine. The cell survival was determined by MTT assay.
After AMT, the ratio of F4/80+ to CD45+ cells in corneas increased after AMT as compared with T group. In vitro BM cells showed decreased proliferative response and cell survival when these cells were co-cultured with AM alone, or in combination with LPS or IFN-gamma. Addition of apoptotic or necrotic cells significantly increased proliferation and survival of BM cells when co-cultured with AM.
After AMT, the ratio of macrophages to bone marrow derived cells (F4/80+/CD45+) in the HSK corneas increases. Under the influence of AM, macrophages proliferate and survive in a highly inflammatory environment after phagocytosis of apoptotic or necrotic cell material.
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