· AIM: To determine the effects of rapamycin on experimental autoimmune uveoretinitis(EAU) and investigate of role of rapamycin on T cell subsets in the disease.·METHODS: EAU was induced in rats using peptid...· AIM: To determine the effects of rapamycin on experimental autoimmune uveoretinitis(EAU) and investigate of role of rapamycin on T cell subsets in the disease.·METHODS: EAU was induced in rats using peptides1169 to 1191 of the interphotoreceptor binding protein(IRBP). Rapamycin(0.2 mg/kg/d) was administrated by intraperitoneal injection for a consecutive 7d after immunization. Th1/Th2/Th17 cytokines, TGF-β1, and IL-6produced by lymphocyteswere measured by ELISA, while Th17 cells and CD4 +CD25 + regulatory T cells(Tregs)from rat spleen were detected by flow cytometry.·RESULTS: Intraperitoneal treatment immediately after immunization dramatically ameliorated the clinical course of EAU. Clinical responses were associated with reduced retinal inflammatory cell infiltration and tissue destruction. Rapamycin induced suppression of Th1/Th2/Th17 cytokines, including IFN-γ, IL-2, IL-17, IL-4, and IL-10 release from T lymphocytes of EAU rats, in vitro.Rapamycin also significantly increased TGF-β1production but had no effect on IL-6 productionof T lymphocytes from EAU rats in vitro. Furthermore,rapamycin decreased the ratio of Th17 cells/CD4 +T cells and upregulated Tregs in EAU, as detected by flow cytometry.·CONCLUSION: Rapamycin effectively interferes with T cell mediated autoimmune uveitis by inhibiting antigen-specific T cell functions and enhancing Tregs in EAU.Rapamycin is a promising new alternative as an adjunct corticosteroid-sparing agent for treating uveitis.展开更多
AIM: To determine the differences of amino acid(AA) levels in experimental autoimmune uveoretinitis(EAU). METHODS: AA analysis of the plasma samples in EAU rats induced by interphotoreceptor retinoid-binding protein e...AIM: To determine the differences of amino acid(AA) levels in experimental autoimmune uveoretinitis(EAU). METHODS: AA analysis of the plasma samples in EAU rats induced by interphotoreceptor retinoid-binding protein emulsion were performed with high performance liquid chromatography(HPLC) and phenylisothiocyanate(PITC) pre-column derivation methods were performed. Using partial least squares discriminant analysis(PLS-DA), the potential biomarkers were identified in EAU rat plasma, and the metabolic pathways related to EAU were further analyzed. RESULTS: The method results showed that linear(r≥0.9957), intra-day reproducible [relative standard deviation(RSD)=0.04%-1.33%], inter-day reproducible(RSD=0.06%-2.07%), repeatability(RSD=0.03%-0.89%), stability(RSD=0.05%-2.48%) and recovery(RSD=1.98%-4.39%), with detection limits of 0.853-11.4 ng/mL. The metabolic profile in EAU rats was different from that in the control groups five AAs concentrations were increased and nine AAs were reduced. Moreover, five metabolic pathways were related to the development of EAU. CONCLUSION: The developed method is a simple, rapid and convenient for determination of AAs in EAU rat plasma, and these findings will provide a comprehensiveinsight on the metabolic profiling of the pathological changes in EAU.展开更多
Osteopontin(OPN)is an extracellular matrix protein with a diverse range of functions,including roles in cell adhesion,migration,and immunomodulation,which are associated with the modulation of neuroinflammation in the...Osteopontin(OPN)is an extracellular matrix protein with a diverse range of functions,including roles in cell adhesion,migration,and immunomodulation,which are associated with the modulation of neuroinflammation in the central nervous system.The present study was performed to evaluate the involvement of OPN in the eyes of an experimental autoimmune uveoretinitis(EAU)model.The EAU model was developed by immunization of Lewis rats with interphotoreceptor retinoid-binding protein.The results showed the OPN level was remarkably upregulated in the eye of EAU rats on day 9 post-immunization.The level of CD44,a ligand of OPN,was increased in the ciliary body of EAU rats.Furthermore,OPN was also detected in the ciliary body and activated microglia/macrophages in the EAU retina.The results suggest that OPN was significantly upregulated in the eyes of EAU rats,and that it may be useful as an early biomarker of ocular autoimmune diseases.All animal experiments were approved by the Institutional Animal Care and Use Committee of Jeju National University(approval No.2020-0012)on March 11,2020.展开更多
To investigate the cellular phenotype and apoptosis of infiltrating cells involved in experimental autoimmune uveoretinitis (EAU) Methods Immunohistochemical staining and in situ apoptosis staining were performed us...To investigate the cellular phenotype and apoptosis of infiltrating cells involved in experimental autoimmune uveoretinitis (EAU) Methods Immunohistochemical staining and in situ apoptosis staining were performed using monoclonal antibodies to monocytes and macrophages (ED1), major histocompatibility complex (MHC) class Ⅱ antigen (OX6), T lymphocytes (R73) and TACS 1 Klenow kit on both ocular sections and wholemounts from Lewis rats after immunization with interphotoreceptor retinoid binding protein (IRBP) Results EAU was induced in 12 of 16 Lewis rats with a mean clinical inflammation score of 1 29±0 7 Influx of monocytes, lymphocytes and MHC class Ⅱ positive cells into the uvea and retina was noted after immunization with IRBP Apoptosis of infiltrating cells was observed in the uvea and retina, and more apoptotic cells were present in the iris and ciliary body compared with those in the choroid and retina Conclusion Apoptosis of infiltrating cells occurs at the early stage of EAU, which may greatly contribute to the rapid regression of the inflammation induced by IRBP展开更多
目的:大鼠自身免疫性葡萄膜视网膜炎(experimentalautoimmune uveoretinitis,EAU)大鼠模型眼部研究细胞因子信号抑制因子(suppressor of cytokine signaling,SOCS)的表达。方法:用光感受器间维生素A类结合蛋白(interphotoreceptorretino...目的:大鼠自身免疫性葡萄膜视网膜炎(experimentalautoimmune uveoretinitis,EAU)大鼠模型眼部研究细胞因子信号抑制因子(suppressor of cytokine signaling,SOCS)的表达。方法:用光感受器间维生素A类结合蛋白(interphotoreceptorretinoid-binding protein,IRBP)免疫Lewis鼠160只后,在眼组织切片上应用SOCS多克隆抗体进行免疫组织化学染色,观察其在眼组织中的表达并与正常大鼠40只做对照。结果:用IRBP免疫Lewis鼠后,免疫组织化学染色发现在虹膜、睫状体和视网膜部位可见SOCS-1和SOCS-5蛋白表达;而在正常Lewis鼠未见SOCS蛋白表达。统计学结果显示,SOCS-1和SOCS-5蛋白表达与EAU严重程度呈正相关(r1=0.954,r2=0.963,P<0.01)。结论:自身免疫性葡萄膜视网膜炎Lewis鼠出现SOCS阳性表达细胞在EAU的发病过程中起了一定的作用。展开更多
基金Supported by National Natural Science Foundation of China(No.81371005)
文摘· AIM: To determine the effects of rapamycin on experimental autoimmune uveoretinitis(EAU) and investigate of role of rapamycin on T cell subsets in the disease.·METHODS: EAU was induced in rats using peptides1169 to 1191 of the interphotoreceptor binding protein(IRBP). Rapamycin(0.2 mg/kg/d) was administrated by intraperitoneal injection for a consecutive 7d after immunization. Th1/Th2/Th17 cytokines, TGF-β1, and IL-6produced by lymphocyteswere measured by ELISA, while Th17 cells and CD4 +CD25 + regulatory T cells(Tregs)from rat spleen were detected by flow cytometry.·RESULTS: Intraperitoneal treatment immediately after immunization dramatically ameliorated the clinical course of EAU. Clinical responses were associated with reduced retinal inflammatory cell infiltration and tissue destruction. Rapamycin induced suppression of Th1/Th2/Th17 cytokines, including IFN-γ, IL-2, IL-17, IL-4, and IL-10 release from T lymphocytes of EAU rats, in vitro.Rapamycin also significantly increased TGF-β1production but had no effect on IL-6 productionof T lymphocytes from EAU rats in vitro. Furthermore,rapamycin decreased the ratio of Th17 cells/CD4 +T cells and upregulated Tregs in EAU, as detected by flow cytometry.·CONCLUSION: Rapamycin effectively interferes with T cell mediated autoimmune uveitis by inhibiting antigen-specific T cell functions and enhancing Tregs in EAU.Rapamycin is a promising new alternative as an adjunct corticosteroid-sparing agent for treating uveitis.
基金Supported by the National Natural Science Foundation of China (No.81373826 No.81674032)+1 种基金Natural Science Foundation of Shandong Province (No. ZR2014HQ074)Key Development & Research Program of Shandong Province (No.2017GSF19110)
文摘AIM: To determine the differences of amino acid(AA) levels in experimental autoimmune uveoretinitis(EAU). METHODS: AA analysis of the plasma samples in EAU rats induced by interphotoreceptor retinoid-binding protein emulsion were performed with high performance liquid chromatography(HPLC) and phenylisothiocyanate(PITC) pre-column derivation methods were performed. Using partial least squares discriminant analysis(PLS-DA), the potential biomarkers were identified in EAU rat plasma, and the metabolic pathways related to EAU were further analyzed. RESULTS: The method results showed that linear(r≥0.9957), intra-day reproducible [relative standard deviation(RSD)=0.04%-1.33%], inter-day reproducible(RSD=0.06%-2.07%), repeatability(RSD=0.03%-0.89%), stability(RSD=0.05%-2.48%) and recovery(RSD=1.98%-4.39%), with detection limits of 0.853-11.4 ng/mL. The metabolic profile in EAU rats was different from that in the control groups five AAs concentrations were increased and nine AAs were reduced. Moreover, five metabolic pathways were related to the development of EAU. CONCLUSION: The developed method is a simple, rapid and convenient for determination of AAs in EAU rat plasma, and these findings will provide a comprehensiveinsight on the metabolic profiling of the pathological changes in EAU.
基金supported by the National Research Foundation of Korea,No.NRF-2019R1A2C1087753(to TS)。
文摘Osteopontin(OPN)is an extracellular matrix protein with a diverse range of functions,including roles in cell adhesion,migration,and immunomodulation,which are associated with the modulation of neuroinflammation in the central nervous system.The present study was performed to evaluate the involvement of OPN in the eyes of an experimental autoimmune uveoretinitis(EAU)model.The EAU model was developed by immunization of Lewis rats with interphotoreceptor retinoid-binding protein.The results showed the OPN level was remarkably upregulated in the eye of EAU rats on day 9 post-immunization.The level of CD44,a ligand of OPN,was increased in the ciliary body of EAU rats.Furthermore,OPN was also detected in the ciliary body and activated microglia/macrophages in the EAU retina.The results suggest that OPN was significantly upregulated in the eyes of EAU rats,and that it may be useful as an early biomarker of ocular autoimmune diseases.All animal experiments were approved by the Institutional Animal Care and Use Committee of Jeju National University(approval No.2020-0012)on March 11,2020.
文摘To investigate the cellular phenotype and apoptosis of infiltrating cells involved in experimental autoimmune uveoretinitis (EAU) Methods Immunohistochemical staining and in situ apoptosis staining were performed using monoclonal antibodies to monocytes and macrophages (ED1), major histocompatibility complex (MHC) class Ⅱ antigen (OX6), T lymphocytes (R73) and TACS 1 Klenow kit on both ocular sections and wholemounts from Lewis rats after immunization with interphotoreceptor retinoid binding protein (IRBP) Results EAU was induced in 12 of 16 Lewis rats with a mean clinical inflammation score of 1 29±0 7 Influx of monocytes, lymphocytes and MHC class Ⅱ positive cells into the uvea and retina was noted after immunization with IRBP Apoptosis of infiltrating cells was observed in the uvea and retina, and more apoptotic cells were present in the iris and ciliary body compared with those in the choroid and retina Conclusion Apoptosis of infiltrating cells occurs at the early stage of EAU, which may greatly contribute to the rapid regression of the inflammation induced by IRBP
