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S. Camelo, A.S. Voon, S. Bunt, P.G. McMenamin; Localisation and Characterisation of Antigen-bearing Cells in the Spleen and Lymph Nodes after Intracameral Injection . Invest. Ophthalmol. Vis. Sci. 2003;44(13):729.
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Purpose: Injection of antigen (Ag) into the anterior chamber (AC) of the eye induces a unique form of immune tolerance termed anterior chamber associated immune deviation (ACAID). It has been proposed that in the murine model of ACAID induction depends on the migration of F4/80+ ocular antigen presenting cells from the eye to the spleen and other lymphoid organs where they deliver a tolerogenic Ag-specific signal. The purpose of the present study is to determine the location and the nature of the Ag-bearing cells in the secondary lymphoid organs following an intracameral injection of fluorescent-labelled Ag. Methods: Twenty-four hours after in vivo single intracameral injection of cascade blue-labelled Dextran (CB-D) (70kD) into the right eye of 8-11 week-old Lewis female rats, animals were perfusion fixed and submandibular lymph nodes (SMLN), cervical lymph nodes (CLN) and spleen were collected. The phenotype of Ag+ cells in lymphoid tissues was determined by immunofluorescence using a range of monoclonal antibodies specific for macrophages (ED1, ED2, ED3), Dendritic cells (ED5, OX6, OX62), T cells (TCR, CD4, CD8), NK cells (CD161) and B cells (CD1, CD45RA, RLN-9D3) and tissues were analysed by confocal microscopy. To determine whether individual cells in the spleen and lymph nodes can trap blood-derived Ag and/or ocular-derived Ag, CB-D was injected into the right eye and FITC-Dextran in the tail vein. Results: Twenty-four hours after injection in the AC, Ag is located in marginal zone macrophages and subcapsular sinus macrophages (ED1+, ED3+, ED5-, OX62- and OX6-) in the spleen and predominantly in the ipsilateral SMLN and right and left CLN. In the spleen, these CB-D-bearing cells are in close association with marginal zone B cells and NK cells. After ocular injection of FITC-Dextran and tail vein injection of CB-D, double positive cells for both Ags are found in the right SMLN and CLN and the spleen. Conclusions: Our data supports the hypothesis that Ag injected in the AC travels from the eye to the secondary lymphoid organs through the blood in a soluble form and is internalized by marginal zone macrophages in the spleen and subcapsular sinus macrophages in the SMLN and CLN. These cells, important in presentation of bacterial and viral T-independent Ag to B cells, may be involved in the specific ocular immune responses and in the induction of ACAID.
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