Immunohistochemical staining of immunoglobulin G in healthy equine, canine, and feline corneas

OBJECTIVE: Establishing an immunohistochemical approach for semi‐quantitative assessment of the presence of immunoglobulin G (IgG) in equine, canine, and feline corneas. PROCEDURES: Healthy corneas of horses, dogs, and cats, euthanized because of a fatal disease or an unrecoverable trauma unrelated to and without a history of ophthalmic disease were formalin‐fixed, paraffin‐embedded, and determined to be pathomorphologically healthy by light microscopy. Automated immunohistochemistry was performed using primary antibodies against IgG, biotin‐conjugated secondary antibodies, and streptavidin‐peroxidase, as well as diaminobenzidine for visualization. After counterstaining with hematoxylin, epithelium, stroma, Descemet´s membrane (DM), and endothelium were semi‐quantitatively scored for the presence of IgG on a 4‐grade scale (0 = no, 1 = faint, 2 = medium, 3 = strong staining) by light microscopy. RESULTS: Corneal specimens of 20 horses (40 eyes) with a median age of 15.5 years (range 2–31 years), 12 dogs (21 eyes) with a median age of 10.0 years (range 4–16), and 13 cats (24 eyes) with a median age of 10.0 years (range 2–18) were included in the study. Different sexes and breeds were represented. In all corneas (100%), significant medium signal intensity in the stroma was observed. Variable immunosignal was obtained in epithelium, DM, and endothelium. CONCLUSION: This method reproducibly allows for the detection of IgG in healthy equine, canine, and feline corneas, particularly stroma. Semi‐quantitative results evidence medium presence of IgG in the corneal stroma. Further research is needed to evaluate IgG presence in diseased corneas.