CD4＋T细胞与非感染性葡萄膜炎

目前认为非感染性葡萄膜炎是由T细胞（主要是CD4＋T细胞）介导的自身免疫性炎症。CD4＋T细胞分为Th1、Th2、Th17和调节性T细胞（Treg）4个亚群,各细胞亚群间通过复杂的细胞因子网络介导自身免疫反应和炎症反应。对各细胞亚群及其在非感染性葡萄膜炎中作用的研究是国内外葡萄膜炎发病机制研究的热点。就CD4＋T细胞各亚群分泌的细胞因子、相互作用及其在非感染性葡萄膜炎中的作用进行综述。     

Selected markers of T cell activation in children with idiopathic uveitis

The aim of our investigation was to assess T cell activation markers (CD69, HLA-DR, CD25, CD49a) in idiopathic uveitis. T cells from children aged 9-17 with idiopathic uveitis were studied. Monoclonal antibodies against chosen markers of T cell activation were used. Expression of the CD69, HLA-DR, CD25, CD49a were determined by flow-cytometry. HLA-DR and CD49a--marker of the late activation--were significantly increased.     

间充质干细胞对实验性自身免疫性葡萄膜炎眼部CD4+T细胞亚群浸润的影响

目的 观察大鼠骨髓间充质干细胞(MSCs)对大鼠实验性自身免疫性葡萄膜炎(EAU)的治疗效果.方法 Lewis大鼠12只随机分为对照组与治疗组,均为6只大鼠.大鼠单后足及背部6点注射含30 μg光感受器间维生素A类结合蛋白多肽片段R16的乳剂0.2 ml,建立EAU模型,观察各组临床表现.贴壁培养法纯化Wistar大鼠骨髓MSCs.建模后第9～11天,治疗组每日行尾静脉注射浓度为5×106/ml的MSCs悬液1 ml,对照组注射等体积磷酸盐缓冲液.建模后第15天,分离各组大鼠眼部细胞,通过流式细胞仪检测干扰素-γ、白细胞介素-17和叉状头螺旋转录因子3阳性细胞的比例.采用混合线性模型进行重复测量资料的方差分析统计临床评分;独立样本t检验统计流式细胞检测结果.结果 建模后第6天,虹膜血管轻度扩张充血;第9天前房轻度混浊,瞳孔缩小,对光反射消失;第12天前房混浊,瞳孔膜闭,眼底红光反射变暗甚至消失;之后炎症反应逐渐减轻.第11～15天,MSCs治疗组临床评分明显低于对照组,差异有统计学意义(t=2.42、2.21、4.16、5.24、4.03,P＜0.05).流式细胞仪检测结果显示,MSCs治疗可以显著减少眼部CD4-T细胞、Th1细胞和Th17细胞比例以及增加调节性T细胞比例.结论 MSCs治疗可以减轻EAU的临床表现;减少CD4+T细胞向眼部浸润,下调眼部效应性Th1细胞、Th17细胞以及上调调节性T细胞.     

葡萄膜炎患者外周血CD4+CD25high调节性T细胞的表达

目的:通过检测葡萄膜炎患者外周血中CD4+CD25high调节性T细胞的表达水平,初步探讨其在葡萄膜炎发病中的作用。方法:取16例正常健康对照及30例不同类型活动期葡萄膜炎患者,采取抗凝外周血,进行三色免疫荧光染色、流式分析。结果:葡萄膜炎患者外周血中CD4+CD25highT细胞的百分率与正常健康对照(8.56%±4.22%vs5.72%±3.11%)相比,显著升高(P=0.022),而且与患者病情严重程度明显相关(P=0.007)。另外CD4+CD25highCTLA4+T细胞在葡萄膜炎患者外周血中比例较正常对照明显升高(P=0.012)。结论:葡萄膜炎患者外周血中CD4+CD25highTreg细胞的异常表达可能参与疾病的发生发展,并与疾病的活动密切相关。     

调节T细胞与葡萄膜炎的研究进展

葡萄膜炎是一种常见的眼科疾病,近年来越来越多的研究证实免疫学机制在其发病中占到重要作用。CD4+CD25+调节T细胞是调节性T细胞的重要亚群之一,对维持机体正常免疫耐受具有重要作用,它的数量与功能变化和许多与免疫相关疾病的发生、发展有关。现就CD4+CD25+调节T细胞与葡萄膜炎的研究进展作一综述。     

葡萄膜炎患者外周血CD4^＋CD25^high调节性T细胞的表达

目的：通过检测葡萄膜炎患者外周血中CD4^＋CD25^high调节性T细胞的表达水平,初步探讨其在葡萄膜炎发病中的作用。 方法：取16例正常健康对照及30例不同类型活动期葡萄膜炎患者,采取抗凝外周血,进行三色免疫荧光染色、流式分析。 结果：葡萄膜炎患者外周血中CD4^＋CD25^highT细胞的百分率与正常健康对照（8.56%±4.22%vs5.72%±3.11%）相比,显著升高（P=0.022）,而且与患者病情严重程度明显相关（P=0.007）。另外CD4^＋CD25^highCTLA4＋T细胞在葡萄膜炎患者外周血中比例较正常对照明显升高（P=0.012）。 结论：葡萄膜炎患者外周血中CD4^＋CD25^highTreg细胞的异常表达可能参与疾病的发生发展,并与疾病的活动密切相关。     

Retinal changes in intermediate uveitis

The authors characterize the clinical unit of intermediary uveitis. They give an account of their own experience with the diagnosis and treatment of this diseases and summarize contemporary views on its aetiopathogenesis and therapy.     

中间葡萄膜炎的超声生物显微镜检查

目的 探讨中间葡萄膜炎超声生物显微镜（ultrasound biomicroscopy,UBM)的图像特征及其应用价值。方法 采用UBM对21例（35只眼）经临床眼科及双目间接检眼镜结合巩膜压迫法检查确诊为中间葡萄膜炎患者行睫状体平坦部、周边部视网膜及玻璃体检查,并对其图像进行分析。结果 31只眼（88．6％）可见病理改变,4只眼（11．4％）未见异常。UBM图像主要表现：周边部玻璃体呈点状混浊、团块状混浊、膜样混虫及睫状体平坦部玻璃体视网膜粘连牵拉征象和睫状体脱离。结论 UBM在中间葡萄膜炎诊断中有临床应用价值。     

Adalimumab specifically induces CD3(+) CD4(+) CD25(high) Foxp3(+) CD127(-) T-regulatory cells and decreases vascular endothelial growth factor plasma levels in refractory immuno-mediated uveitis: a non-randomized pilot intervention study

Aim

To explore immunoregulatory and anti-inflammatory pathways specifically targeted by a subcutaneous anti-TNFαdrug—adalimumab—which might be relevant for controlling refractory uveitis.

Design

Non-randomized pilot intervention study on the effects of adalimumab on Treg populations and plasma VEGF levels in refractory uveitis patients. Inflammatory and immunological parameters were measured in 12 patients before therapy, and 1 and 6 months after therapy, and analyzed in the context of ophthalmological outcomes. The results were compared with those obtained in 10 systemic prednisone-treated uveitis patients.

Results

After 1 month of treatment, all patients responded, with 67% of adalimumab group and 80% of the corticosteroid group achieving inactivity (P=0.5). Unlike steroid-treated patients, a significant increase in T-regulatory CD4⁺ CD25^high Foxp3⁺ CD127⁻ cells was observed in adalimumab patients after 1 month of treatment, and maintained after 6 months (P=0.003). A significant adalimumab-specific drop in plasma VEGF was observed after 1 and 6 months of treatment (P=0.019). In every single patient, Tregs but not VEGF correlated with disease activity.

Conclusions

In refractory uveitis patients treated with adalimumab, clinical efficacy may be mediated through upregulation of Tregs in addition to modulation of VEGF-mediated inflammatory pathways. These biological properties, which were not observed in patients treated with corticosteroids, may reflect the specificity of TNF-αtargeting.     

Immune responses to retinal self-antigens in CD25(+)CD4(+) regulatory T-cell-depleted mice

PURPOSE: Prior work has shown that autoimmune uveoretinitis develops spontaneously in CD25(+)CD4(+) regulatory T-cell-depleted mice (Tr-depleted mice). In this study, the generation of autoantibodies and autoreactive T-cells specific to retinal antigens was examined in Tr-depleted mice with uveoretinitis, and the pathogenic and immunogenic abilities of the autoreactive T cells were evaluated. METHODS: Tr-depletion was achieved in (C57BL/6 x A/J) F1 (B6A) mice by thymectomy on day 3 of life followed by intraperitoneal injection of an anti-CD25 mAb. At 6 months of age, autoantibodies to the retina were evaluated by indirect immunofluorescence, and total IgG2a levels in sera were assessed by ELISA. The pathogenic abilities of the splenic T cells were examined by adoptive transfer to syngeneic nu/nu mice, and the proliferation responses and the secretion of granulocyte-macrophage-colony-stimulating factor (GM-CSF), IFN-gamma, and IL-10 on stimulation by retinal self-antigens was also evaluated. RESULTS: Autoantibodies to the retinal photoreceptor cell layer were detected in Tr-depleted mice, and the titers correlated well with the grades of inflammatory lesions. The splenic CD4(+) T cells of Tr-depleted mice induced uveoretinitis in the recipients by adoptive transfer and exhibited proliferative responses and secretion of IFN-gamma, but not of IL-10, by in vitro stimulation with S-Ag and interphotoreceptor retinoid-binding protein (IRBP). Moreover, the total IgG2a level in serum was markedly and significantly augmented in Tr-depleted mice. CONCLUSIONS: The results suggest that in Tr-depleted mice in which uveoretinitis develops, S-Ag and IRBP-specific T cells are spontaneously sensitized and shifted to a Th1-phenotype. These sensitized T cells may account for the development of autoimmune uveoretinitis in Tr-depleted mice.     

CD69 expression on peripheral CD4+ T cells parallels disease activity and is reduced by mycophenolate mofetil therapy in uveitis

PURPOSE. To assess the effects of mycophenolate mofetil (MMF) therapy on T helper cell activation status, using CD69 expression and cytokine profile with flow cytometry in relation to clinical activity in uveitis. METHODS. Patients with posterior or intermediate uveitis treated with MMF (n = 10), patients with active uveitis not treated with MMF and receiving no or minimal therapy (n = 10), and healthy volunteers (n = 21) had peripheral blood lymphocyte immunofluorescence analysis for T helper cell (CD4, CD3) markers, activation status (CD69), and intracellular cytokine (interleukin [IL]-2, interferon [IFN]-gamma, and IL-4) levels. Patients were compared before and during MMF therapy in relation to T helper cell activation and clinical activity. RESULTS. Patients with active uveitis not treated with MMF and receiving no or minimal therapy had increased frequency of CD69-positive CD4 T cells (10.5% +/- 4.6%, P = 0.0007) compared with healthy volunteers (3.3% +/- 2.7%). Of all patients receiving MMF therapy, only patients with moderate to severe uveitis activity in the pre-MMF treatment group (n = 5; 15.5% +/- 5.0%, P = 0.004) had increased frequency of CD69-positive CD4 T cells compared with healthy volunteers. During MMF therapy, a significant reduction in frequency of CD69-positive CD4 T cells occurred in patients with prior moderate to severe uveitis activity (to 8.9% +/- 3.8%, P = 0.04). Levels of CD69-positive CD4 T cells in patients who had had inactive or mildly active disease (n = 5) before and during MMF therapy were comparable with levels in healthy volunteers. No significant changes in cytokine levels were found between the patient and control groups. A significant association between changes in frequency of CD69-positive CD4 T cells and changes in visual acuity (P = 0.008) and changes in vitreal haze (binocular indirect ophthalmoscopy score; P = 0.01) was observed in MMF-treated patients with prior moderate to severe uveitis activity. CONCLUSIONS. Reduction in uveitis activity during MMF therapy correlates with reduction in frequency of peripheral blood CD69-positive CD4 cells. The frequency of CD69-positive CD4 T cells is a measure of activity in posterior uveitis and may guide adequate immunosuppression.     

Ultrasound biomicroscopic imaging in intermediate uveitis

BACKGROUND—Clinical examination of the region of the eye mainly affected in patients with intermediate uveitis is difficult and often hampered by media opacities. In that perspective ultrasound biomicroscopy (UBM) promises to be a valuable additional diagnostic tool.
METHODS—UBM was performed at a sound frequency of 50 MHz on 26 eyes of 13 patients with intermediate uveitis in order to determine configuration of pars plana, peripheral retina, and vitreous. Findings of ophthalmoscopy with scleral indentation and UBM were compared.
RESULTS—In 18 of 26 eyes pathological structures such as membraneous or fluffy vitreous condensations were identified by UBM. Among these UBM revealed pathological findings which were not visible on funduscopy in nine eyes. Most importantly, vitreoretinal adhesions with traction on the retina were imaged in four eyes. However, in three eyes vitreous opacities being visible on funduscopy were not identified by UBM.
CONCLUSION—UBM seems to be a valuable diagnostic technique for the evaluation of patients with intermediate uveitis. Longitudinal studies will have to determine the relevance of UBM findings for the individual clinical course and their influence on therapeutic decisions.

Keywords: intermediate uveitis; ultrasound biomicroscopy; vitreous opacities     

Fluorescein angiography findings in intermediate uveitis

The present paper describes the results of angiographic examinations of 48 eyes (29 patients) with intermediate uveitis. More than 50% of the cases displayed pathologic changes of the retinal blood vessels, such as increased fluorescein staining of the vessel walls and leakages of the retinal veins or venules, respectively. Some 20% of the eyes manifested cystoid macular edema and/or edema of the optic disk which had gone undetected by ophthalmoscopy. These findings suggest that vascular changes may play a role in the pathogenesis of intermediate uveitis. The question as to whether this disease might be caused rather by retinal vasculitis than by uveitis is discussed.     

CD56+ T cells in the peripheral blood of uveitis patients

AIMS—Natural killer T (NKT) cells, T lymphocytes expressing both T cell and NK cell markers, are suggested to be involved in autoimmune diseases. To examine the relation between the pathogenesis of uveitis and CD56+ T cells, which are thought to be a type of human NKT cells, we investigated peripheral CD56+ T cells in uveitis patients.METHODS—41 uveitis patients (Behçet''s disease (BD), 14; sarcoidosis (SAR), eight; Vogt-Koyanagi-Harada disease (VKH), five; idiopathic uveitis (IU), nine; and others, five) and 19 healthy controls participated in this study. Cell surface antigens of lymphocytes were analysed by use of monoclonal antibodies and flow cytometry.RESULTS—The proportion of CD56+ T cells in patients with BD was higher than in controls and in patients with SAR, VKH, IU, and others.CONCLUSION—Increased peripheral CD56+ T cells might be relevant to the pathogenesis of uveitis in BD, and increase of peripheral CD56+ T cells may be one of the laboratory findings to suggest that uveitis originates from BD.     

Long-term follow-up of intermediate uveitis in children

PURPOSE: To analyze the clinical manifestations, remissions, and visual prognosis of intermediate uveitis in children, and to identify the risk factors for poor visual outcome. DESIGN: Retrospective cohort study. METHODS: Institutional study of 32 consecutive patients examined at a tertiary referral center with intermediate uveitis and the onset of ocular disease before the age of 16 years. Numerous variables were assessed, including age and gender distribution, laboratory data, the presence of systemic diseases, onset and course of ocular inflammation, clinical features and complications, therapeutic strategies and their outcomes, remission and final visual acuity, and characteristics associated with poor visual outcome. RESULTS: Bilateral involvement was observed in 94% of the patients. Remission was observed in seven out of 15 patients (47%) with completed follow-up of five years. For our 32 subjects, we found a mean time to remission of 6.4 years (SE 0.7, CI 5.1 to 7.7). Visual outcome was favorable as only three patients developed unilateral acuity of less than 0.1 after five-year follow-up, and no additional blind eyes manifested. No associated systemic diseases were established. Optic disk edema was the most frequent complication observed (71%). Cystoid macular edema (CME) was observed in 44% of the patients and was the most common cause of visual loss. CONCLUSIONS: Intermediate uveitis of childhood might exhibit a self-limiting course after several years. Visual loss was limited despite the high rate of severe ocular complications.