Objective To observe the therapeutic effect of mensenchymal stem cells (MSCs) for experimental autoimmune uveitis (EAU). Methods MSCs were obtained from Wistar rats and selected by plastic adherence. Lewis rats were divided into treatment group and control group, six rats in each group. EAU models were induced by immunization with an emulsion (0.2 ml) containing 30 mu;g interphotoreceptor retinoid-binding protein derived peptide R16 and complete Freundprime;s adjuvant. The clinical manifestations of two groups were observed. Nine to 11 day after modeling, 1 ml MSCs suspension, which contained 5times;106 MSCs, were injected into the rats in treatment group via tail vein, and the rats in control group were given equal volume of phosphate buffer solution. Fifteen day after modeling, the eyes were collected to test the proportion of interferon gamma;, interleukin-17 and Foxp3 positive cells by flow cytometry. The clinical scores were analyzed by mixed linear model and statistical analysis of variance of repeated measurement data. The results of flow cytometry were analyzed using independent-sample t test. Results Six days after immunization, mild dilatation and congestion of iris vascular was observed. Nine days after immunization, mild muddy anterior chamber, myosis and absent pupillary reaction to light were observed. Twelve days after immunization, muddy anterior chamber, occlusion of pupil and dimmed or disappeared red reflex were observed, and then inflammation was slowly reduced. From 11 to 15 days after immunization, the clinical score of treatment group was lower than that in control group, the difference was statistically significant (t=2.42, 2.21, 4.16, 5.24, 4.03; P＜0.05). The results of flow cytometry showed that MSCs treatment could decrease the proportion of CD4＋T cells, Th1 cells and Th17 cells, increase the proportion of Treg cells. Conclusion MSCs treatment can ameliorate EAU, up-regulate the expression of Treg cells and down-regulate the expression of CD4＋T cells, Th1 cells and Th17 cells.
Objective To observe the characteristics of optical coherence tomography (OCT) of the macular in uveitis patients. Methods A total of 51 cases (78 eyes) of uveitis admitted to affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine from May 2011 to May 2012 were reviewed retrospectively. All patients were underwent OCT examination and classified according to characteristics of OCT. Results The characteristics of OCT in uveitis were classified into six groups. Macular edema including cystoid macular edema and diffuse edema was detected in 48 eyes (61.5%). Macular epiretinal membrane was found in 22 eyes (28.2%). Choroidal neovascularization and macular hole was found in 4 eyes (5.1%), respectively. Ten eyes (12.8%) showed macular atrophy and 22 eyes (28.2%) showed serous neuroepithelium detachment. Conclusions Macular OCT in uveitis patients displayed different morphological characteristics.
Objectives To investigate the clinical characteristics and prognosis of syphilitic uveitis. Methods Clinical charts of 32 syphilitic uveitis patients were retrospectively analyzed. The diagnosis was confirmed by clinical and laboratory tests. There were 32 patients (50 eyes), 18 males and 14 females; the ages were from 21 to 62 years ole, with a mean age of 42 years old. Eighteen patients were bilateral. All patients had complete ocular examinations including visual acuity, intraocular pressure, slit-lamp biomicroscopy, ophthalmoscopy, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA). Results Inflammatory cells in the anterior chamber and corneal endothelium were present in 42 eyes. Thirty eyes showed congestion and swelling of optic discs. Yellowwhite lesions in the posterior pole were present in 18 eyes. No change in 6 eyes. FFA showed staining or hyperfluorescence of optic disc in 32 eyes, venous leakage in 34 eyes, and cystoid macular edema in 15 eyes．ICGA showed squamous or disseminative hypofluorescence damages in 26 eyes. All patients were treated with penicillin and glucocorticoids, 36 eyes had improved vision and fundus damage had abated. Conclusions Most syphilitic uveitis was panuveitis with retinal vasculitis. The prognosis is good with early diagnosis and timely treatment of this disease．
Objective To investigate the effect of peptidoglycan (PGN) on the secretion of pro-inflammatory cytokines by dendritic cells (DCs) and the regulation of T helper 17 (Th17) responses in experimental autoimmune uveitis. Methods Bone marrow cells from naive mice were cultured with granulocyte macrophage-colony-stimulating factor and interleukin (IL)-4 to induce DCs. DCs cultured for six days were randomly divided into two groups: PGNtreated group and control group. The DCs in PGNtreated group were stimulated with PGN and the same volume of phosphate buffered saline was added to the DCs as control group. The relative mRNA expression levels of IL-23, tumor necrotic factor alpha; (TNF-alpha;), IL-6,IL-1beta;were measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Peptide fragment of interphotoreceptor retinoidbinding protein (IRBP1-20)specific T cells, which were isolated from the spleen and draining lymph nodes of C57BL/6 mice immunized with IRBP1-20 peptide fragments 13 days earlier, were co-cultured with PGN-treated or untreated DCs, respectively. Total RNA from T cells cocultured for two days were isolated and the relative expression of retinoic acid receptor-related orphan receptor gamma;t (ROR-gamma;t), IL-17, T-box expression in T cells (T-bet), interferon gamma; (IFN-gamma;) mRNA were detected by realtime RT-PCR. On the second, the fifth and the seventh day, the cocultured T cells were analyzed by flow cytometry to detect the percentages of IFN-gamma;, IL-17 positive cells. Results The real-time RT-PCR results revealed that the level of IL-23, IL-1beta;, IL-6, TNF-alpha; mRNA from PGNstimulated DCs were significantly increased compared to the control group (t=-14.363, -5.627, -3.85, -28.151; P＜0.05). The level of RORgamma;t, IL-17 mRNA from the T cells cocultured with PGN-stimulated DCs were greatly increased compared with the control group (t=-5.601, -19.76；P＜0.05). However, the level of T-bet, IFN-gamma; mRNA from the T cells cocultured with PGNstimulated DCs were significantly decreased compared with the control group (t=4.717, 11.207; P＜0.05). Data of flow cytometry showed that at two days, five days, seven days after cocultured with PGN-treated DCs, the percentages of IL-17 positive T cells were increased compared to the control group (t=-2.944, -3.03, -4.81; P＜0.05), and the percentages of IFN-gamma; positive T cells had no remarkable change (t=-1.25, -0.18, -2.16; P＞0.05). Conclusion PGN can promote the secretion of Th17-related cytokines by DCs, which favors proliferation and differentiation of Th17 in experimental autoimmune uveitis.
Objective To investigate the impact of bone marrow mesenchymal stem cell transplantation on a rat model of experimental autoimmune uveitis (EAU) and analyze its immune regulatory mechanisms in vivo.Methods Eighteen Lewis rats were randomly divided into three groups: model control group, intervention group and normal control group, six animals in each group. Human retinal S-antigen peptide (HS-AgP35, 1 mg/ml) was mixed and emulsified with complete Freundprime;s adjuvant and injected into hind foot pad of rats on the first and eighth day to establish the animal model of EAU. For bone marrow mesenchymal stem cell transplantation, 1 ml of cell suspension (2times;106 cells/ml) was injected into tail vein of the intervention group rats on the first day when the emulsified S-antigen was injected. EAU manifestation, pathological change and IFN-gamma; level were evaluated and compared among those three groups after two weeks. Results No abnormal signs were found in the eyes of rats in normal control group. The manifestation grading of the intervention group (two rats at grade 0, three rats at grade 0.5, one rat at grade one) was significantly different from the model control group (one rat at grade one, one rat at grade two, three rats at grade three, one rat at grade four) (P=0.015). The retina of rats in normal control group was ordinary under light microscope. The histopathologrical grading of the intervention group (one rat at grade 0, four rats at grade 0.5,one rat at grade one) and the model control group (four rats at grade three, two rats at grade four) was also statistically different (P＜0.01). Furthermore, the IFN-gamma; level in peripheral blood of the intervention group rats declined significantly compared to the model control group (t=9.0574, P=0.01). Conclusions Bone marrow mesenchymal stem cells can inhibit EAU significantly, possibly by lowering the level of IFN－gamma;, thereby reduce the severity of uveitis and improve the condition of uveitis in rats.
Objective To analyze the onset, clinical manifestation, causation, complications of pediatric uveitis. Methods One hundred and two patients with uveitis under 16 year-old were retrospectively studied. They visited the clinic in Peking University First Hospital from November 1979 to December 2008. Their age ranged from 2.5 to 16 years old, with a mean of 11.9 years. Routine exam was carried out, including visual acuity, slit lamp, fundus, and laboratory workup. The diagnosis and classification were made by the anatomic location according to the standard of The International Uveitis Working Group. The data of disease history, age of onset, manifestation, recurrence, causation, systemic diseases, complications, and lab examination were analyzed.Results A total of 102 patients (170 eyes) with pediatric uveitis were included in this study, 68 patients (66.6%) were bilateral cases. Anterior uveitis represented in 38 patients (37.3%), intermediate uveitis in 19 (18.6%), posterior uveitis in 10 (9.8%), and panuveitis in 35 (34.3%). The disease duration was from five days to 2.4 years, with a mean of 3.6 months. The follow-up time was two weeks to more than ten years. The first three causes of pediatric uveitis were juvenile idiopathic arthritis, Vogt-Koyanagi-Harada disease, and Behccedil;etprime;s disease. 36 patients were found with complications, and among them 19 had complicated cataract, seven had secondary glaucoma, five had corneal band dystrophy, 12 had iris synechia (both anterior and posterior), one had retinal detachment, two had eye atrophy, and one patient with juvenile idiopathic arthritis had bilateral femoral head necrosis because of the use of steroid and hip joint was replaced. There were ten children suffering more than two complications. Conclusions Pediatric uveitis is a possible blindness disease with variety of etiology and manifestations,and tends to cause complications. Early and special attention must be taken to avoid serious consequences.
Objective To examine the role of recombinant interleukin-10 (IL-10) and the therapeutic effect of endotoxin-induced uveitis (EIU) in rats.Methods Fifty-six male Wistar rats were randomized into three groups. IL-10 treatment group and positive control group had 24 rats respectively, and the normal control group had eight rats. Endotoxin-induced uveitis (EIU) is an established animal model of acute ocular inflammation induced by LPS intravenous injection (1 mu;g/kg). The onset times and signs were observed and the clinical scores were recorded. The blood samples and the aqueous humor samples of right eye were collected separately before the rats were sacrificed at fourth hour, 24th hour and third day after LPS injection. The enzyme-linked immunosorbent assay was used to measure tumor necrosis factor (TNF) alpha;,IL-6, and IL-10 levels in the serum and aqueous humor. The left eyes were used for pathological examination and pathological grading. Results The symptoms of uveitis were appeared in all 24 rats in the positive group. The average onset time was (3.81plusmn;1.05) hours, the average clinical score was 3.67plusmn;1.97. The mild manifestations of uveitis were also appeared in all of the rats in treatment group. The average onset time was (5.63plusmn;1.02) hours, the average clinical score was 2.00plusmn;1.25. The average onset time in treatment group was postponed compared with the rats of positive group (t=4.95, P=0.000). The clinical scores (t=3.50, P=0.00) and the pathological grades (t=3.28, P=0.00) in treatment group were lower than those of positive group. There were not signs or pathologic changes in all the eight rats in the negative control group. The serum and aqueous humor levels of TNF-alpha; and IL-6 in the rats of positive group were higher than those of the treatment group and control group (F=15.34, 57.65, 67.59, 8.42; P=0.00). The serum and aqueous humor levels of IL-10 in the rats of treatment group were higher than those of the positive group and the control group (F=17.84,7.76; P=0.00). There were positive correlations between the level of aqueous humor TNF-alpha;, serum and aqueous humor levels of IL-6 and the disease severity (reye＝0.58, 0.31,0.81, rpath＝0.56, 0.31, 0.74; P＜0.05). The negative correlations were presented between the serum levels of IL-10 with the disease severity (r=-0.54,-0.55; P=0.00). There were negative correlations between the serum and aqueous humor levels of TNF-alpha; and IL-6 and the onset time of the disease (r＝-0.47,-0.59,-0.77,-0.36; P＜0.05) as well. Conclusions These findings bly suggest that suppressive IL-10 is a potent candidate for the prevention of TNF-alpha; and IL-6 in uveitis and could be applied as a novel immunoregulatory agent to control EIU.
The knowledge of uveitis of Chinese eye doctors has been improved in general. While the usage of glucocorticoid agents was more reasonable, other non-corticoid immunosuppressant get more attention recently. The usage of antibiotics also has being reduced gradually. The international impact of our uveitis research has been enhanced. However there are still some problems, such as big difference between different regions of uveitis research, still many misunderstandings on the treatment of uveitis complications, and the reasonable evaluation of intravitreal injection with glucocorticoid needs emphasis. In China Behcetprime;s disease and Vogt-Koyangi-Harada syndrome are the most common uveitis subtypes which can lead to blindness,but some rare subtypes of uveitis are also increasing such as syphilitic uveitis, acquired immune deficiency syndrome（AIDS），mycotic endophthalmitis and masquerade syndrome. In the future we need cooperative studies between multicenters to investigate the effectiveness of different treatment strategies for Behcetprime;s disease and Vogt-Koyangi-Harada syndrome, and to optimizing the best therapeutic schedule. We also need to pay more attentions to the clinical features of those uveitis subtypes which increased recently；and to investigate the prevention and therapeutic effect of induction of immune tolerance to uveitis.
Objective To examine the levels of interferon-gamma; (INF-gamma;), tumor necrosis factor-alpha; (TNF-alpha;) and interleukin-6(IL-6) in serum of patients with acute uveitis before and after treatment, and to explore the possible roles of those cytokines in the initiation and progression of the uveitis. Methods A series of 75 patients with acute uveitis,and 30 healthy persons from our hospital were investigated. The levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase and convalescent phase were measured by the enzymelinked immunosorbent assay. Result The serum levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase were significantly higher than that of the convalescent phase and the healthy controls (F=65.805/50.418/155.381, P=0.000). A significant negative correlation was found between the serum levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase with their initial visual acuity(r=-0.656, -0.592 and -0.653, Plt;0.01). There was also a positive correlation among the serum levels of INF-gamma;, TNF-alpha; and IL-6(r=0.340, 0.467 and 0.338, Plt;0.05). Conclusions There are high serum levels of INF-gamma;, TNF-alpha; and IL-6 in patients with acute uveitis, and the cytokines levels were decreased after the treatment. The results suggested that the INF-gamma;, TNF-alpha; and IL-6 involved in initiation and progression of uveitis.
Objective To observe the proportion changes of CD4+CD25+FOXP3+ T cells in peripheral blood of patients with VogtKoyanagiHarada disease (VKH) before and after one month of treatment. Methods he peripheral blood samples from 15 patients with VKH disease before and after one month of treatment by glucocorticoid, and from 15 healthy volunteers were collected，and lymphocytes were separated from them. CD4+CD25+ regulatory T cells were labeled by antibodies of cell surface marker CD4、CD25 and transcription factor FOXP3. The proportion of CD4+CD25+FOXP3+ T cells were detected by flow cytometry. Results Before the treatment, the percentage of CD4+CD25+FOXP3+ T cells in periphery blood was（0.30plusmn;0.19）% of CD4+ cell in VKH patients, and（1.41plusmn;0.52）% in control group, the difference was statistically significant（t=7.665,Plt;0.01）; after one month of treatment, the VKH patients group was（1.28plusmn;0.54）% which close to the control group. However there were two patients whose CD4+CD25+ T cells increased extraordinarily after one month of treatment. Conclusions The proportion of CD4+CD25+ FOCP3+ T cells in periphery blood in VKH patients were lower than control group obviously before treatment, but were close to control group after treatment. Those results indicated that VKH diseases may be associated with the decreased proportion of CD4+CD25+ regulatory T cells.