Oftalʹmologiâ (Oct 2014)
Immune mapping of the peripheral part of the visual analyzer and optic nerve
Abstract
Aim. To perform immune mapping of the peripheral part of visual analyzer and optic nerve in order to identify potential antigenic targets of autoimmune attack. Methods. Eyes enucleated for terminal painful glaucoma (n = 30) were studied. Immunohistochemistry (IHC) was performed on paraffin-embedded sections of isolated retina and optic nerve using a broad panel of antibodies, i.e., monoclonal murine anti-MBP (myelin basic protein) antibodies, polyclonal rabbit anti-alpha fodrin antibodies, monoclonal murine anti-NSE2 (neuron-specific enolase) antibodies, monoclonal murine anti-GFAP (glial fibrillary acidic protein), and polyclonal rabbit anti-S100 antibodies. IHC reaction was visualized using Mouse and Rabbit Specific HRP / AEC Detection IHC Kit. IHC reaction without primary antibodies included was a negative control. IHC reaction was considered as follows: negative — no specific cellular staining or less than 10 % of cells are stained; mild — 10‑30 % of cells are stained (+); moderate — 30‑75 % of cells are stained (++); marked — more than 75 % of cells are stained (+++); overexpression — 100 % of cells intensively express markers. Additionally, staining intensity was considered as mild (+1), moderate (+2), strong (+3) and intense (+4).Results. Immune mapping with a broad panel of monoclonal antibodies identified ocular structures which were stained with IHC markers. Retina was stained with almost all markers of neural differentiation (i.e., antibodies against NSE, GFAP, S100, and α-fodrin) excepting anti-MBP autoantibodies. IHC reaction intensity in retinal layers and structures varied and depended on markers. Moderate (2+) staining with antibodies against MBP, NSE, GFAP, and S100 and marked (3+) staining with antibodies against alpha-fodrin was detected in the cytoplasm of optic nerve glia.Conclusion. Complete labelling of retina structures was performed. As a result, IHC profiles of retinal neurons, optic nerve axons, interneurons, and microglial cells were described. IHC profiles of retinal layers and optic nerve are useful markers which can be applied in serological diagnostics of various ocular disorders.
