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1 From the Department of Ophthalmology–Immunology and the 2 Department of Morphology, Netherlands Ophthalmic Research Institute, Amsterdam; the 3 Department Ophthalmology, University of Aberdeen, United Kingdom; the 4 Department Anatomy and Human Biology, University of Western Australia, Perth; the 5 Department Ophthalmology, University of Amsterdam, The Netherlands.
PURPOSE. Previous studies have shown that experimental autoimmune encephalomyelitis (EAE) and anterior uveitis (AU) develop in Lewis rats immunized with myelin basic protein (MBP). The purpose of this study was to characterize the dynamics, distribution, and phenotype of infiltrating cells in the iris during EAE-associated AU.
METHODS. Lewis rats were immunized with MBP emulsified in complete Freund’s adjuvant (CFA) or with CFA alone. Cellular infiltration of the iris was analyzed at various time points by immunohistochemistry of wholemounts, flow cytometry, and immunoelectron microscopy, by using monoclonal antibodies specific for monocytes/macrophages (ED1), T lymphocytes (R73, W3.25, OX8), T-cell activation markers (OX39, OX40), granulocytes (HIS48), major histocompatibility complex (MHC) class II (OX6), and neurofilament (2H3).
RESULTS. MBP-immunized rats showed development of characteristic monophasic EAE, followed, after resolution of paralysis, by mild self-limited AU. Initially, focal infiltrates of round MHC class II+ and ED1+ cells were found in the iris. During the course of AU, the midiris became massively infiltrated with ED1+ monocytes-macrophages, R73+ T cells, granulocytes (HIS48+), and MHC class II+ cells. The influx of T cells consisted of CD4+ and CD8+ cells, of which only a small fraction (<14 and 11%, respectively) expressed activation markers. The infiltrating cells accumulated in proximity to myelinated and nonmyelinated nerve bundles and in the vicinity of blood vessels in the iris. No evidence was found for demyelination or nerve degradation. Neither EAE nor AU developed in CFA-treated control rats.
CONCLUSIONS. These data show that EAE-associated AU is characterized by a transient mixed cellular infiltrate consisting of monocytes-macrophages, granulocytes, and CD4 and CD8 T cells. The preferential accumulation of inflammatory cells in the vicinity of nerve fibers suggests that AU in this model may result from autoreactivity to nerve antigens.
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