动物造模,动物葡萄膜视网膜炎 z-bio 2024-01-29 849

　　1. Animal model production: Six short tailed monkeys weighing 3-4 kg, regardless of gender.

　　(1) Preparation of S antigen: S antigen is isolated and purified through three steps from retinal extracts isolated from human retina. ① Precipitation in semi saturated ammonium sulfate; ② Filtration through cross-linked dextran gel; ③ Hydroxyapatite chromatography adsorption purification.

　　(2) Induction of experimental active immune uveitis (EAU) using 100 μ Six short tailed monkeys were immunized with retinal S antigen dissolved in complete Freund's adjuvant. Specifically, 0.5ml of soluble S antigen was emulsified with 0.5ml of complete Freund's adjuvant, and 1ml of prepared retinal S antigen was subcutaneously injected into 5 areas between the scapulae. Before injection, slit lamp, indirect ophthalmoscopy, and fluorescence angiography (FFA) examinations did not show any signs of inflammation, nor did they detect retinal vascular occlusion or fluorescence leakage. Ketamine (15-25mg/kg) intramuscular injection anesthesia was administered before eye and vascular imaging examinations.

　　2. Observation indicators and results: Eye slit lamp and indirect ophthalmoscopy should be checked once a week to guide angiography, and should be continuously performed within 5 months after active immunomodulatory modeling. Among the 6 animals, 3 underwent retinal laser photocoagulation before modeling. In the nearly 9-month observation of the course of the disease, it was found that regardless of the severity of uveitis, there was a sudden worsening of the condition one month after modeling. Both eyes did not develop the disease simultaneously, but one eye developed the disease 1-5 weeks after the other eye. Specific signs included retinal vessel (vein and small vein) occlusion, optic disc edema, anterior uveitis, subretinal exudation, vitreous inflammation, and occasional anterior uveitis. The most severe symptoms usually occur in three monkeys who have not undergone pre photocoagulation.

　　In summary, all reported uveitis models have been induced by antigens, known as experimental autoimmune uveitis (EAU), which is an organ specific CD4+T cell-mediated intraocular inflammatory disease. It is a commonly used disease model for studying human endogenous uveitis, especially posterior uveitis. The antigens used include superantigens (S2Ag) and interphotoreceptor resin binding protein (IRBP), as the antigens mainly originate from the photoreceptor cell layer, which naturally becomes the main target of EAU's autoimmune response. The inflammation is mainly located in the posterior part of the eyeball, with the posterior uvea being the main focus. In addition to granulomatous proliferation of the retina and choroid, the main lesions are retinal vasculitis and perivascular inflammation. In severe cases, retinal detachment may occur, and the anterior chamber may not be affected or mildly affected.

　　Through different studies by scholars in recent years, animal models of uveitis have also shown a trend of diversification. Experimental animals include rabbits, guinea pigs, rats, and primates, and the success rates of different animal models also vary. Although mouse EAU models are more effective than