ABSTRACT
AIM: To investigate the cytotoxic effect of specific T cells from mice with experimental autoimmune uveitis (EAU) as well as their secreted interferon (IFN)-γ and interleukin (IL)-17A on murine photoreceptor (661W) cells. METHODS: An EAU model was established in female mice by injection of interphotoreceptor retinoid binding protein (IRBP) emulsion supplemented with complete Freund's adjuvant (CFA) and Mycobacterium tuberculosis (TB). On day 12 after induction of EAU, specific T cells from spleen and lymph node tissues were isolated and cultured for 4d and the levels of IFN-γ and IL-17A in the supernatants were determined by enzyme-linked immunosorbent assays (ELISAs). T cells and their supernatants were added to 661W cells to observe the alteration of cell morphology; IFN-γ and IL-17A were separately added to 661W cells to observe the effect of IFN-γ and IL-17A on cell proliferation. RESULTS: The levels of IFN-γ and IL-17A in the T cell supernatants were 1568.64±38.79 pg/mL and 1456.57±46.98 pg/mL, respectively. The supernatants apparently inhibited 661W cell proliferation (P<0.05). T cells could also attach to the surface of 661W cells, and IFN-γ showed a more serious cytotoxic effect on 661W cells than IL-17A, inhibiting cell proliferation (P<0.01). CONCLUSION: IFN-γ and IL-17A from T cells of EAU mice model can exert cytotoxic effects on murine photoreceptor cell proliferation, and IFN-γ shows more serious cytotoxic effects on murine photoreceptor cells than IL-17A.
ABSTRACT
OBJECTIVES: To observe the potential protective effect of angiopoietin-1 (Ang-1) on rat choroidal neovascularization (CNV) leakage. METHODS: The study was conducted at the Eye Institute of Shandong University of Traditional Chinese Medicine, Jinan, China from June 2012 to June 2013. Thirty CNV model rats were induced by laser. In vivo, fluorescein fundus angiography and pathological techniques were applied to detect the effect of vascular endothelial growth factor (VEGF) and Ang-1 intravitreous injection. In vitro, 3-(4, 5-dimethylthiazole-2-yl)-2, 5-biphenyl tetrazolium bromide (MTT) assay was applied to detect the proliferation of cultured bovine retinal endothelial cells (BRECs) after treatment with VEGF and Ang-1. Transmission electron microscopy (TEM) was used to detect the morphological changes under VEGF and Ang-1. RESULTS: In the CNV rat model, less late leakage was found in the Ang-1 group than the vehicle control or the VEGF group. The MTT assay showed Ang-1 administration inhibited the proliferation of BRECs. The VEGF promoted proliferation at low concentrations and inhibited the proliferation when its concentration reached 50 ng/ml. The administration of VEGF+Ang-1 rescued the inhibition effect of Ang-1 alone. The TEM results showed that there were less intercellular junctions in the VEGF group compared with the vehicle control. In the VEGF + Ang-1 group, the intercellular junctions were nearly normal. CONCLUSION: The Ang-1 can induce intercellular junction formation and decrease the CNV leakage.