Animal Model Of Eales Disease Induced By Intravitreal Injection Of Purified BCG-PPD Protein

Objective:To establish animal model of Eales disease induced by the intravitreal injection of purified BCG protein derivative.Methods:(1) Grouping and Sensitization method:Adult male SD rats (n=75) were randomly divided into five groups. Experimental group Ⅰ, Ⅱ, Ⅲ, Ⅳ, and control (n=15in each group). Experimental group I:Equal volume of lmg/ml HS-Agp35and FICA (Eph type incomplete adjuvant) was put into a bowl to be mixed and emulsifie then taking0.1ml antigen to inject into one foot of rats intracutaneously, and injecting BCG-PPD5U and FICA into the other side of the foot intracutaneously to sensitize the rats. BCG-PPD5U was injected intravitreally after14days.0.2ml antigen was taken for experimental group Ⅱ. In experimental group Ⅲ, Ⅳ, and Ⅴ, sensitization method was the same as experimental group Ⅰ.HS-Agp35lOUg was injected intravitreally after14days for experimental group Ⅲ. Experimental group IVwas blank control (nothing was given after14days). Experimental V group was given intravitreally0.9%NS0.1ml after14days.(2)Observational index:clinical observation, fluorescein fundus angiography (FFA), spread of retina, retinal pathology, measuring retinal VEGF, PDGF and helper T lymphocytes factor in the blood of Rat model and to make correlation analysis.Results:1. Clinical observation:(1) there was obvious uveitis reaction in Experimental group Ⅲ.(2) surface neovascularization can be founded in individual rat of Experiment group Ⅰ and Ⅱ at three months after iris.(3) experimental group Ⅳ and Ⅴ (control group) had no positive signs.2. FFA:(1). There was intraocular inflammation mainly on the base of retinal vascular disease in Experiment group Ⅰ and Ⅱ.(2). There was intraocular inflammation on the base of choroidopathy disease in Experimental group Ⅲ.(3) There were no positive signs to be found in Experimental group IV, V group (control group).3.Spread of retina:(1). There was intraocular inflammation on the base of retinal vascular in Experiment group Ⅰ and group Ⅱ, but vasculitis degrees of two groups can not be distinguished morphologically.(2). There was intraocular inflammation on the base of choroidopathy disease in Experimental group Ⅲ, but inflammation can affect retinal blood vessels.(3) There were no positive signs to be found in Experimental group IV, V group (control group).4.(1) Immunohistochemistry (VEGF):each group had40samples, and the obtained positive area of experimental group was group Ⅰ> group Ⅱ> group Ⅲ.(2) Immunohistochemistry (PDGF):each group had40samples, there was positive results in Experimental group Ⅰ, and there were not positive results in Experimental group Ⅱ and Ⅲ.5. Retinal pathology:the results showed the thickening, folds, structural disorder, retinal vascular dilation, congestion of retina in experimental group Ⅰ, Ⅱ, and Ⅲ, and that there were irregular internal limiting membrane, a small amount of vascular endothelial cells in a breakthrough internal limiting membrane, and cell proliferation under the internal limiting membrane, disorganized in the group Ⅰ and Ⅱ. In group Ⅰ, retinal neovascular was occasionally seen in the front or surface of the retina, and in experimental group III, it can be observed that cells in the internal limiting membrane arranged in slightly irregular.6. Determination of helper T lymphocytes factor in the blood of Rat model:(1) the IFN-γ Concentration content of serum respectively in the rat model group and in the normal control group were (385.16±19.72)(200.38±15.84),TNF-α were (25.75±2.04)(12.14±1.16),IL-4were (13.18±0.34)(11.56±0.36) Concentration of serum IFN-γ, TNF-α, IL-4in the rat model group is higher compared with normal control group (P<0.01);(2)The ratio of IFN-γ/IL-4respectively in the rat model group and in the normal control group were (29.20±1.29)(17.35±1.35), the ratio in the rat model group is higher compared with normal control group(P<0.01).Conclusion:1. After foot of rats was intracutaneously sensitized with single HS-Agp35, the combination of HS-Agp35and BCG-PPD antign, intravitreal injection of BCG-PPD can cause inflammation of retinal blood vessels, and the retinal vasculitiscloser of United sensitized group, which is more similar to human Eales disease;2. The BCG-the PPD plays a leading role in Eales disease model caused by the United sensitization;3. In Eales disease model induced by sensitized rats through intravitreal injection of BCG-the PPD, the imbalance of T lymphocyte subsets is involved in the immune mechanisms of Eales disease;...