April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Amniotic Membrane Induces Peroxisome Proliferator-activated Receptor Positive Alternatively Activated Macrophages
Author Affiliations & Notes
  • Dirk Bauer
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Maren Hennig
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Susanne Wasmuth
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Martin Busch
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Hanna Baehler
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Klaus-Peter Steuhl
    Department of Ophthalmology, University of Duisburg-Essen, Essen, Germany
  • Solon Thanos
    Institute of Experimental Ophthalmology, School of Medicine, University Muenster, Muenster, Germany
  • Arnd Heiligenhaus
    Department of Ophthalmology at, St. Franziskus Hospital, Ophtha-Lab, Munster, Germany
  • Footnotes
    Commercial Relationships  Dirk Bauer, None; Maren Hennig, None; Susanne Wasmuth, None; Martin Busch, None; Hanna Baehler, None; Klaus-Peter Steuhl, None; Solon Thanos, None; Arnd Heiligenhaus, None
  • Footnotes
    Support  DFG Grants Ba 2248/1-1, Ba 2248/1-2, He 1877/12-2; and Ernst and Berta Grimmke Foundation
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1128. doi:
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      Dirk Bauer, Maren Hennig, Susanne Wasmuth, Martin Busch, Hanna Baehler, Klaus-Peter Steuhl, Solon Thanos, Arnd Heiligenhaus; Amniotic Membrane Induces Peroxisome Proliferator-activated Receptor Positive Alternatively Activated Macrophages. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1128.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Amniotic membrane transplantation (AMT) reportedly improves herpetic stromal keratitis (HSK). Here we studied the role of the AM on macrophages in this process.

Methods: : BALB/c mice with necrotizing HSK received an AMT or tarsorrhaphy (TAR) as control. Apoptosis of corneal macrophages was determined by flow cytometric analysis using annexin V/7-AAD system. Macrophage invasion was determined by using a cornea invasion assay. Cytokine secretion was determined by ELISA. Arginase activity was determined by bioassay. Expression of MAPK, Akt, NFkB, or PPAR-gamma related proteins was determined by western blot, and the expression of costimulatory surface molecules or ppar-gamma by flow cytometry. Lipid accumulation was observed by Oil red O and Sudan B staining.

Results: : After AMT, macrophages in the HSK cornea had apototic features. and AM treated corneas had increased macrophage invasion. IL-6, IL-10, IL-12, TNF-alpha, and NF-kB content in HSK corneas had decreased with AMT. AMT increased expression of P-PTEN, P-GSK-3b, P-C-Raf, P-p44/42-MAPK, PPAR-gamma, arginase1+2, and arginase activity in AM treated HSK corneas, while P-Akt and P-MSK expression was decreased. In vitro, NF-kB, cytokine and chemokine production, costimulatory molecules (CD80, CD86, CD40), phagocytic capacity, proliferation, viability, and accessory function to HSV-1 specific T lymphocytes where reduced in AM conditioned bone marrow-derived macrophages (BM), while CD206, CD204, CD163, CD68, lipid accumulation in the cytoplasm, PPAR-gamma expression, and arginase activity was increased. An increase in viability and proliferation was observed in the presence of AM combined with apoptotic cells, as compared to AM alone, and this was associated with decreased PPAR-gamma and CD36 expression.

Conclusions: : We conclude that the improvement in HSK lesions after AMT is associated with increased macrophage cell death, and the modulation of classically activated macrophages into alternatively activated macrophages. Apoptotic cells induced in the environment with AM support the presence and survival of such macrophages.

Keywords: herpes simplex virus • inflammation • wound healing 
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