Expression of CC chemokines and their receptors in the eye in autoimmune anterior uveitis associated with EAE.

PURPOSE: To determine the pattern of expression of CC chemokines and their receptors in the eyes of Lewis rats and to establish their role in autoimmune anterior uveitis (AU) associated with experimental autoimmune encephalomyelitis (EAE). METHODS: EAE/AU was induced in Lewis rats with myelin basic protein in complete Freund's adjuvant (CFA). The rats were scored for the development of clinical EAE and AU. The expression of CCL5/regulated on activation normal T-cell expressed and secreted (RANTES), CCL2/monocyte chemotactic protein (MCP)-1, CCL3/macrophage inflammatory protein (MIP)-1alpha, and CCL4/MIP-1beta and their receptors was examined at the preclinical stage, onset, peak, and recovery by RT-PCR and ELISA. EAE/AU rats were treated with neutralizing polyclonal antibodies against CCL3/MIP-1alpha, CCL4/MIP-1beta, CCL2/MCP-1, and CCL5/RANTES and tested for the suppression of onset of clinical AU and EAE. The control group received normal rabbit IgG at the same dose. RESULTS: The gene expression of those chemokines was upregulated concurrently with symptom onset of EAE/AU and correlated with the intensity of inflammatory changes in the eye and central nervous system (CNS). The highest expression of CCL4/RANTES, CCL2/MCP-1, and CCL3/MIP-1alpha in the eye was detected at onset of clinical uveitis, whereas CCL4/MIP-1beta was elevated at the peak of AU. The expression of chemokine receptors associated with T-helper (Th)1-type response, CCR1 and CCR5, correlated with their appropriate ligands and was the highest at the peak of AU, whereas CCR2, the receptor for CCL2/MCP-1, was present before the onset of the disease. Treatment of anti-MIP-1beta and anti-MCP-1 significantly delayed the onset and shortened the duration of AU and EAE. Anti-MIP-1alpha treatment had no effect on clinical EAE but inhibited the clinical signs of AU. Although CCL5/RANTES expression was observed during the entire course of the disease, anti-RANTES treatment had no effect on clinical disease progression. CONCLUSIONS: The data suggest that CCL2/MCP-1, CCL3/MIP-1alpha, and CCL4/MIP-beta contribute to the recruitment of inflammatory cells into the eye and CNS and to disease activity.     

The role of T cells in autoimmune uveitis

Autoimmune diseases result from the activation of self-reactive T cells recognizing autoantigens or foreign antigens cross-reactive with an autoantigen. T cells are thought to play a major role in autoimmune diseases in different organs, including the eye. This review focuses on the role of T cells in autoimmune uveitis in humans and in animal models of experimental autoimmune uveitis. Since rheumatoid arthritis is an autoimmune disease that has been studied far more extensively than uveitis, we have also included a review of clinical and experimental observations relevant to that disease.     

Systemic management of autoimmune anterior uveitis

Autoimmune uveitis is an acute, recurrent, sight-threatening disease that can lead to severe visual loss and blindness. It requires systemic immunosuppressive therapy and continuous monitoring by ophthalmologist and rheumatologist. There are no universally accepted referral patterns for the treatment of endogenous uveitis. Most specialists indicate corticosteroids, methotrexate and cyclosporine A as the first use drugs. Anti-cytokine drugs are a new opportunity for the patients unresponsive to the conventional anti-inflammatory and immunosuppressive therapy.     

Granulomatous anterior uveitis associated with bimatoprost

Purpose: To describe a previously unreported case of anterior granulomatous uveitis in a patient using bimatoprost. Methods: A 72-year-old woman with a long-standing history of anisometropic amblyopia and pseudoexfoliative glaucoma in the right eye started therapy with bimatoprost 0.03% once a day in the right eye. She had no previous history of ocular inflammation or ocular surgery. Her medical history was negative for systemic diseases associated with ocular inflammation. Results: After one week, the patient developed severe conjunctival injection, cells and flare, and numerous ‘mutton fat’ keratic precipitates in the right eye. Examination of the left eye revealed no evidence of inflammation. Bimatoprost was discontinued; no topical steroid therapy was started. Systemic investigations were normal. The inflammation resolved over two weeks, solely with the discontinuation of bimatoprost. Conclusions: Bimatoprost is a synthetic prostamide, chemically related to prostamide F. Prostamides are naturally occurring substances, biosynthesized from anandamide in a pathway that includes COX2. Even though anandamide has proven suggestive potential pro-inflammatory effects, the mechanism of induction of inflammation by bimatoprost remains uncertain and speculative. In our report, the onset of acute uveitis in a patient using bimatoprost, after a long-term and well-tolerated treatment with a prostaglandin analog, suggests a distinct potential pro-inflammatory action of prostamides. This can indirectly support the concept that the target receptor of bimatoprost is different, and that the mechanism of action of prostamides is pharmacologically unique.     

Treatment of autoimmune anterior uveitis with recombinant TCR ligands

PURPOSE: To determine protective properties of recombinant TCR ligands (RTLs) as a new treatment for experimental autoimmune anterior uveitis (AU). RTLs comprise the rat RT1.B beta1alpha1 domains, linked either to the guinea pig MBP69-89 peptide (RTL201), to the corresponding rat MBP69-89 peptide (RTL200), or to the cardiac myosin peptide CM-2 (RTL203). METHODS: AU associated with experimental autoimmune encephalomyelitis (EAE) was actively induced in Lewis rats by injection of myelin basic protein emulsified in complete Freund's adjuvant (CFA) or passively by the transfer of pathogenic T cells. Rats received five daily doses each of 300 microg RTL201 in saline, intravenously. Control rats received the same dose of RTL203 or an "empty" beta1alpha1 protein (no peptide). The rats were evaluated for the suppression of clinical and histologic signs of AU. RESULTS: RTL201 prevented active and passive AU and reduced the clinical symptoms of established AU. RTL201 completely prevented clinical and histologic AU in the treated rats, compared with disease progression in the untreated rats or those treated with an "empty" construct. The suppression of clinical AU correlated with a significant reduction in inflammatory cells infiltrating the eyes of the RTL201-treated rats. Furthermore, RTL201 inhibited T cell proliferation, DTH responses, and cytokine mRNA expression in the eye, in contrast to the untreated rats. In comparison with RTL201, RTL200 was less effective in protecting the eye from AU. RTL203 also significantly inhibited clinical AU, but not EAE. CONCLUSIONS: RTL constructs suppressed clinical and histologic AU by inhibiting the systemic activation of specific T cells and preventing the recruitment of inflammatory cells into the eye. These findings suggest a possible clinical application of this novel class of peptide/MHC class II constructs in patients with AU that is mediated by T-cell responses to known antigenic peptides.     

Bilateral nongranulomatous anterior uveitis associated with bimatoprost

A 72-year-old man with long-standing bilateral glaucoma became refractory to levobunolol ophthalmic solution therapy after many years. Brimonidine was prescribed, but the patient developed a hypersensitivity several months later that was treated with loteprednol ophthalmic suspension. Bimatoprost was initiated 2 weeks later. Within an hour of the first dose of bimatoprost, the patient reported eye pain and photophobia that remained unresolved the following day. Examination revealed acute bilateral nongranulomatous anterior uveitis that was effectively treated with loteprednol. While observations in human and animal models suggest an association between certain prostaglandin-like agents and intraocular inflammation, this report is one of the first to suggest a link between bimatoprost and intraocular inflammatory reaction.     

Spontaneous hyphema associated with anterior uveitis.

Few reports have described hyphema in association with anterior uveitis. Five cases of anterior chamber haemorrhage are reported in patients with five different anterior uveitic entities: Reiter's syndrome, juvenile chronic arthritis, ankylosing spondylitis, idiopathic anterior uveitis, and herpes simplex. Hyphema has been reported in association with idiopathic non-granulomatous anterior uveitis, but not with the other four entities. In three cases, iris rubeosis was present. In two cases the patients were taking non-steroidal anti-inflammatory agents. The hyphemas occurred at times of heightened inflammation and resolved spontaneously without complication in all but one case, a boy with idiopathic uveitis who required surgery to remove the blood. The clinical outcome of these cases provides evidence that conservative medical management is usually sufficient.     

实验性自身免疫性葡萄膜炎T淋巴细胞亚群与Treg细胞的表达研究

目的　探讨实验性自身免疫性葡萄膜炎（ｅｘｐｅｒｉｍｅｎｔａｌａｕｔｏｉｍｍｕｎｅｕｖｅｉｔｉｓ,ＥＡＵ）新西兰兔模型外周血单个核细胞（ｐｅ-ｒｉｐｈｅｒａｌｂｌｏｏｄｍｏｎｏｎｕｃｌｅａｒｃｅｌｌｓ,ＰＢＭＣ）Ｔ淋巴细胞亚群的变化及其与疾病特征的相关性。方法　选取健康成年雄性新西兰大白兔４０只作为动物模型,用２０ｇ·Ｌ－１牛血清蛋白（ｂｏｖｉｎｅｓｅｒｕｍａｌｂｕｍｉｎ,ＢＳＡ）静脉注射和玻璃体内注射法建立ＥＡＵ模型,并观察模型眼炎症反应及ＰＢＭＣＴ淋巴细胞亚群特征。结果　ＣＤ４＋Ｔ细胞在ＥＡＵ兔的外周血中比例明显增加,且随时间的进展呈进行性增加,并与炎症反应呈正相关,相同的趋势可见于ＣＤ４＋／ＣＤ８＋Ｔ细胞比例。此外,Ｔｒｅｇ细胞趋势与ＣＤ４＋Ｔ细胞及ＣＤ４＋Ｔ／ＣＤ８＋Ｔ细胞比例相反,与炎症反应呈负相关。结论　ＣＤ４＋Ｔ细胞的优势性选择性克隆增殖及Ｔｒｅｇ细胞的降低有可能引发免疫功能紊乱并导致针对自身组织的免疫应答失去有效的负性调控,导致ＥＡＵ的发生及进行性加重。     

Treg细胞在实验性自身免疫性葡萄膜炎发病过程中的动态变化

背景 研究表明,调节性T细胞(Treg)是一类负向调控免疫应答的T细胞亚群,在维持免疫稳态和免疫耐受方面发挥重要作用.自身免疫性葡萄膜炎是一种免疫性眼病,Treg细胞在自身免疫性葡萄膜炎发生和发展过程中的调控作用尚不完全清楚. 目的 观察Treg细胞在实验性自身免疫性葡萄膜炎(EAU)大鼠发病过程中的动态变化.方法 选取84只6～8周龄SPF级Lewis大鼠,采用随机数字表法随机分为模型组和对照组.模型组于大鼠双后足垫、腹部双侧及背部皮下注射光感受器间维生素A类结合蛋白(IRBP)1177-1191、结核菌素(TB)、完全弗氏佐剂(CFA)和PBS混合乳化剂共300μl,对照组大鼠以同样方法皮下注射等容量不含IRBP的TB与CFA乳化剂.分别于造模后第9、13、18、23、28、35、48天观察模型组和对照组大鼠的眼部炎症症状并根据严重程度进行炎症评分.于造模后上述时间点分别处死模型组及对照组大鼠各6只,采用常规组织病理学方法观察各组大鼠眼部虹膜、睫状体和视网膜组织形态变化;分离大鼠脾脏淋巴细胞,采用流式细胞术检测大鼠脾脏悬液中Treg细胞特异性标志物Foxp3标记细胞比例;采用实时荧光定量PCR法检测大鼠脾脏淋巴细胞中Foxp3 mRNA相对表达量变化. 结果 免疫后第8天模型组大鼠虹膜血管扩张充血,开始出现眼部炎症表现,免疫后第13天虹膜血管明显扩张,前房可见渗出和积脓,瞳孔区有膜样渗出,炎症评分最高,为(3.75±0.42)分,之后眼部炎症反应逐渐减轻,至免疫后第23天炎症反应接近消失,造模后第7、11、13、15、17、19、21天间模型鼠眼炎症反应评分总体比较差异有统计学意义(F=81.709,P＜0.001).对照组大鼠眼部检查正常.组织病理学观察发现,模型组大鼠虹膜、睫状体、视网膜等组织有中性粒细胞、淋巴细胞和单核细胞浸润,组织结构排列疏松,以免疫后第13天最为明显,此后逐渐减轻,至免疫后第23天虹膜、睫状体和视网膜组织结构接近正常,炎性细胞浸润消失.流式细胞技术检测发现,免疫后第13、18、23、28、35、48天模型组大鼠脾脏Foxp3标记细胞比例分别为(5.50±0.64)％、(13.36±0.98)％、(10.34±0.79)％、(9.58±1.02)、(6.73±0.81)％和(5.58±0.47)％,明显高于对照组的(2.80±0.38)％、(3.36±0.53)％、(3.65±0.57)％、(3.37±0.43)％、(3.33±0.50)％和(3.13±0.61)％,差异均有统计学意义(t=-6.272、-15.556、-11.910、-9.753、-6.154、-5.491,均P＜0.01).模型组和对照组造模后各时间点大鼠脾脏淋巴细胞中Foxp3 mRNA的相对表达量变化与Foxp3标记细胞比例变化趋势基本一致. 结论 EAU大鼠葡萄膜炎的发病及转归与Treg细胞数量和功能变化密切相关.     

Cytotoxic effect of specific T cells from mice with experimental autoimmune uveitis on murine photoreceptor cells

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.     

通过前房相关免疫偏离阻止实验性自身免疫前葡萄膜炎

目的 实验性自身免疫性前葡萄膜炎（ＥＡＡＵ）是研究人前葡萄膜炎的有用模型。本研究旨在观察前房相关免疫偏离（ＡＣＡＩＤ）能否阻止ＥＡＡＵ发生。方法 将溶于磷酸盐缓冲液（ＰＢＳ）的牛黑色素蛋白（ＢＭＰ）注射于大鼠右眼前房;对照组动物仅注射ＰＢＳ。７d后使用且加完全福氏佐剂（ＣＦＡ）的ＢＭＰ和百日咳毒素免疫所有动物。通过临床 观察和组织病理学检测葡萄膜炎的严重程度和发病率。通过由注射ＢＭＰ刺激的足垫水肿反     

Prevention of experimental autoimmune anterior uveitis by anterior chamber-associated immune deviation

目的实验性自身免疫性前葡萄膜炎（ＥＡＡＵ）是研究人前葡萄膜炎的有用模型。本研究旨在观察前房相关免疫偏离（ＡＣＡＩＤ）能否阻止ＥＡＡＵ发生。方法将溶于磷酸盐缓冲液（ＰＢＳ）的牛黑色素蛋白（ＢＭＰ）注射于大鼠右眼前房;对照组动物仅注射ＰＢＳ。７ｄ后,使用添加完全福氏佐剂（ＣＦＡ）的ＢＭＰ和百日咳毒素免疫所有动物。通过临床观察和组织病理学检测葡萄膜炎的严重程度和发病率。通过由注射ＢＭＰ刺激的足垫水肿反应评估迟发型超敏反应（ＤＴＨ）。结果与对照组相比,预先眼内注射ＢＭＰ的大鼠,其ＥＡＡＵ的严重程度和发病率减低,且ＤＴＨ反应受到显著抑制。结论这些资料提示ＡＣＡＩＤ可以用来阻止ＥＡＡＵ的发生。     

Chronic granulomatous anterior uveitis associated with multiple sclerosis

A retrospective review is presented of the medical records of seven female patients with uveitis associated with definite multiple sclerosis. Six of the patients had severe bilateral chronic granulomatous anterior uveitis, which in four cases resulted in extensive posterior synechiae and scarring of the peripupillary iris. One patient had bilateral intermediate uveitis with unilateral posterior synechiae. Severe chronic granulomatous anterior uveitis associated with multiple sclerosis has a predilection for women, can precede neurological symptoms and can be controlled with topical steroids.