人眼组织中共刺激分子、Fas/FasL及主要组织相溶性复合体类分子的免疫组织化学研究

目的　探讨正常人眼组织 (虹膜睫状体、脉络膜、视网膜 )中共刺激分子 B7- 1和 B7- 2、与凋亡相关的分子 Fas/ Fas L、人类白细胞抗原 ( human leukocyte antigen,HL A) - DR及 CD6 8(抗巨噬细胞抗体 )分子的表达及意义。　方法　 12只正常供体人眼 (死后 16～ 2 4h)均来自中山眼科中心眼库 ,将 6只供体眼组织 (虹膜睫状体、脉络膜、视网膜 )行常规冰冻切片 ,另 6只做视网膜平片 ,分别于各部分眼组织切片及视网膜平片进行免疫组织化学研究 ,探讨正常人眼组织中 B7- 1、B7- 2、HL A- DR、CD6 8、Fas/ Fas L分子的表达。　结果　人虹膜睫状体中有 B7- 2、Fas L、CD6 8、HL A- DR分子的表达 ;脉络膜中有 B7- 1、Fas L、CD6 8、HL A- DR分子的表达 ;视网膜中可见 HL A- DR、CD6 8、Fas L分子的表达 ,而无 B7- 1、B7- 2、Fas分子的表达。　结论　共刺激分子 B7- 1和 B7- 2、Fas/ Fas L及主要组织相溶性复合体 ( major histocompatibility com-plex,MHC- )类分子在不同人眼组织中的表达不同 ,这些分子的表达可能对维持人眼组织免疫环境的稳定性起着重要作用。     

人眼组织中共刺激分子、Fas/FasL及主要组织相溶性复合体II类分子的免疫组织化学研究

目的 探讨正常人眼组织(虹膜睫状体、脉络膜、视网膜)中共刺激分子B7-1和B7-2、与凋亡相关的分子Fas/FasL、人类白细胞抗原(human leukocyte antigen, HLA)DR及CD68(抗巨噬细胞抗体)分子的表达及意义。 方法 12只正常供体人眼(死后16～24 h)均来自中山眼科中心眼库，将6只供体眼组织(虹膜睫状体、脉络膜、视网膜)行常规冰冻切片，另6只做视网膜平片，分别于各部分眼组织切片及视网膜平片进行免疫组织化学研究，探讨正常人眼组织中B7-1、B7-2、HLA-DR、CD68、Fas/FasL分子的表达。 结果 人虹膜睫状体中有B7-2、FasL、CD68、HLA-DR分子的表达；脉络膜中有B7-1、FasL、CD68、HLA-DR分子的表达；视网膜中可见HLA-DR、CD68、FasL分子的表达，而无B7-1、B7-2、Fas分子的表达。 结论 共刺激分子B7-1和B7-2、Fas/FasL及主要组织相溶性复合体(major histocompatibility complex, MHC-II)类分子在不同人眼组织中的表达不同，这些分子的表达可能对维持人眼组织免疫环境的稳定性起着重要作用。 （中华眼底病杂志，2003，19：109-112）     

交感性眼炎的临床与免疫组化分析

通过对因交感性眼炎而摘除的眼球进行病理及免疫组化分析 ,结合临床特征 ,探讨交感性眼炎的发病机理。方法 :17例诊断为交感性眼炎行交感眼眼球摘除的病人 ,眼球标本用HE染色 ,观察炎症细胞在脉络膜、巩膜血管及神经周围的分布及细胞种类。其中 10例进行免疫组化染色 ,用CD3、CD45RO标记T淋巴细胞 ,CD2 0标记B淋巴细胞 ,CD6 8标记巨噬细胞。观察这些抗体在脉络膜、巩膜血管及神经、结膜下血管的分布特点及强度。结果 :HE染色 :淋巴细胞浸润在脉络膜大中血管层及脉络膜上腔 ,间有上皮样及多核巨细胞。巩膜内血管及神经亦可见淋巴细胞浸润。免疫组化 :CD3、CD45RO阳性细胞分布在脉络膜、巩膜、结膜下组织中 ,多在上皮样细胞、巩膜静脉及神经、结膜下血管周围。CD2 0主要分布在脉络膜上腔及外层脉络膜。CD6 8阳性细胞主要分布在脉络膜上皮样细胞、多核巨细胞及巩膜及结膜下血管周围。结论 :交感性眼炎是一种非环死肉芽肿性葡萄膜炎 ,炎症不仅局限于脉络膜 ,亦可见于巩膜和结膜下。巩膜的血管及神经周围存在吞噬色素的上皮样细胞 ,提示交感性眼炎的发生与色素颗粒或其他相关抗原密切相关。     

中心性浆液性脉络膜视网膜病变误诊分析

目的探讨分析中心性浆液性脉络膜视网膜病变误诊为交感性眼炎和其他眼底疾病,误诊为中心性浆液性脉络膜视网膜病变的原因。方法对初诊为交感性眼炎的2例和初诊为中心性浆液性脉络膜视网膜病变的5例患者进行散瞳眼底检查和眼底荧光血管造影,部分行靛青绿血管造影检查,观察病情追踪随访。结果2例初诊为交感性眼炎的患者为中心性浆液性脉络膜视网膜病变;5例初诊为中心性浆液性脉络膜视网膜病变的患者中2例为脉络膜转移癌,1例为老年黄斑变性合并陈旧性大泡性视网膜病变,2例为下方渗出性视网膜脱离伴黄斑浅脱离。结论对眼底病患者要重视散瞳详细检查眼底,对激素引起的色素上皮屏障功能损害,要注意与交感性眼炎相鉴别。     

Behcet病患者外周血淋巴细胞共刺激分子的表达

目的观察Behcet病患者外周血淋巴细胞CD80(B7-1)、CD86(B7-2)、CD28、CTLA-4分子的表达。方法采用流式细胞三色直接免疫荧光标记法，对24例Behcet病患者(Behcet病组)和20个正常人(对照组)外周血T、B淋巴细胞亚群的共刺激分子CD80、CD86、CD28和CTLA-4的表达进行检测。结果Behcet病患者外周血CD ⁴⁺ T细胞表面 CTLA-4 分子的表达[(3.18±1.18)%]高于对照组[(1.73±0.66) %]，两者差异有显著性的意义(t=-3.722,P<0.01)；CD19⁺ B细胞表面CD86分子的表达[(4.49±1.73)%]高于对照组[(2.40±1.49) %]，两组比较差异有显著性的意义(t=－2.071,P<0.05)。比较两组间其它分子的表达，差异均无显著性的意义。结论Behcet病患者 B7、 CD28的相互作用促进了葡萄膜炎的发生，阻断其相互作用可能为今后葡萄膜炎的治疗提供一种新思路。(中华眼底病杂志,2003,19:357-359)     

眼球穿孔伤后的临床免疫学研究

眼穿孔伤常累及晶体,虹膜和睫状体。晶体和色素膜是两种隐蔽抗原,在一定的条件下可能引起变态反应性色素膜炎,甚至引起双眼典型交感性眼炎或晶体过敏性交感性眼炎。随着眼科免疫学研究工作的开展,我们试图在临床免疫学的研究中,了解眼穿孔伤后机体的     

玻璃体切除术后的交感性眼炎

交感性眼炎是一种较少见的双眼肉芽肿性全葡萄膜炎,常并发于眼外伤。特别是累及葡萄膜组织的穿通伤或内眼术后。近年来随着玻璃体手术的广泛开展,由其引起的交感性眼炎已有报道。作者最近遇到一例因裂孔源性视网膜脱离伴增殖性玻璃体视网膜病变。行经睫状体平部玻璃体切除联合巩膜冷凝、环扎术后发生交感性眼炎,现报告如下。患者男57岁因右眼裂孔源性视网膜脱离伴     

经巩膜二极管激光睫状体光凝术后交感性眼炎二例

缺血型视网膜中央静脉阻塞容易发生新生血管性青光眼,睫状体光凝术和冷冻术是治疗新生血管性青光眼的有效方法,关于睫状体光凝或冷冻术后发生交感性眼炎的报告较少见,尤其是二极管激光睫状体光凝术后还未见发生交感性眼炎的报道.我们报告2例如下:     

γ-干扰素对人眼组织共刺激分子及Fas/FasL分子表达的影响

目的 探讨γ-干扰素对人眼组织中共刺激分子及Fas/FasL分子表达的影响。 方法 正常供体人眼9只用于实验。其中6只眼作视网膜平片，每只眼取12片视网膜平片；分别将视网膜平片放于24孔培养板中，共分为3组，每孔含Dulbecco改良Eagle(DMEM)/F12培养液2 ml，3组γ-干扰素（IFN-γ）浓度分别为0、200、1000 U/ml；37℃培养箱中培养(95%O 2，5%CO 2)24h后取出固定，用免疫组织化学方法检测各视网膜平片上B7-1、B7-2、Fas/FasL分子的表达。同时取另外3只正常人眼制作视网膜平片做为正常对照组，直接用免疫组织化学方法检测平片中上述分子的表达。 结果 正常人视网膜平片中有FasL分子的表达，但无B7-1、B7-2和Fas分子表达；当视网膜平片与IFN-γ体外培养后，视网膜中出现B7-1、B7-2和Fas分子的表达，并且FasL分子表达量增多。 结论 IFN-γ可通过诱导共刺激分子及Fas/FasL分子表达参与免疫反应的发生和诱导细胞凋亡，在T淋巴细胞的激活中起着重要作用。 (中华眼底病杂志, 2006, 22: 117-119)     

B7：CD28／CTLA—4共刺激分子及其在眼科的研究进展

大量实验证实B7家族分子与其受体CD28和CTLA-4结合后所产生的共刺激信号在T淋巴细胞激活中起重要作用,决定着受到抗原刺激的T细胞是分化、增殖为效应细胞,还是进入无反应状态,其在眼科的研究主要集中于眼科自身免疫性疾病、角膜和视网膜三个方面,阻断B7：CD28/CTLA-4态。其在眼科的研究主要集中于眼科自身免疫性疾病、角膜和视网膜三个方面,阻断B7：CD28/CTLA-4通路可能为眼科自身免疫性疾病、角膜移植排斥反应和视网膜葡萄膜疾病的治疗提供新的途径,本就其分子特性、功能及其在眼科的研究进展等方面的有关资料作一综述。     

B7+ iris pigment epithelial cells convert T cells into CTLA-4+, B7-expressing CD8+ regulatory T cells

PURPOSE: To determine whether iris PE (IPE) promotes the generation of regulatory T-cells (Tregs) with cell contact via B7-2/CTLA-4 interactions. METHODS: T cells were cocultured with IPE cells obtained from eyes of normal and B7-deficient mice, x-irradiated, and used as regulators. IPE T regulator cells (IPE Tregs) of normal and CD28- or CTLA-4-deficient mice were established. Target bystander T cells were established from normal splenic T cells with anti-CD3 antibodies. T-cell activation was assessed for proliferation by [3H]-thymidine incorporation. Neutralizing anti-B7-1 and/or B7-2 antibodies, anti-CTLA-4 antibodies, CTLA-4-Ig fusion proteins were used to abolish regulatory function. IPE-exposed CD8+ T cells were evaluated for expression of B7, CTLA-4, and Foxp3 by using RT-PCR and flow cytometry. CD8+ IPE Tregs were depleted of B7-2+ and CTLA-4+ T cells and assayed for suppressive activity by adding them to bystander T cells. RESULTS: T cells acquired T regulatory activity when exposed to cultured IPE. Ciliary body PE cells did not promote conversion of T cells into Tregs. IPE converted CD8+, but not CD4+, T cells into Tregs by direct cell contact. In the conversion, IPE and responding T cells must both express endogenously synthesized B7-1 and B7-2, and the T cells must also express CTLA-4. Expression of CD28 molecules was not necessary for Treg generation. In addition, the CD8+ Tregs that fully suppress activation of bystander T cells expressed Foxp3. CONCLUSIONS: IPE cells promote conversion of T cells into Tregs solely through a contact-dependent mechanism. T cells exposed to IPE cells acquire full regulatory capacity.     

Mooren ulcer: an immunopathologic study

PURPOSE: To determine the pattern and distribution of mononuclear cells, adhesion, and co-stimulatory molecules in the conjunctiva of patients with Mooren ulcer. METHODS: Conjunctival biopsy specimens were obtained from 6 patients with Mooren ulcer and 6 healthy individuals. Immunohistochemistry was performed on frozen sections of the cryopreserved human conjunctivas using monoclonal antibodies directed against CD1alpha, CD3, CD4, CD8, CD20, CD25, CD57, and CD68 cells; the adhesion molecules E-selectin, vascular cell adhesion molecule-1 (VCAM-1), very late activation-4 (VLA-4), ICAM-1, and LFA-1; and the co-stimulatory molecules CD28, B7-1, B7-2, and CTLA-4. RESULTS: Differences in expression on the conjunctival epithelium from patients with Mooren ulcer and normal subjects were noted only for VCAM-1, VLA-4, ICAM-1, and LFA-1. The ratio of CD4+/CD8+ cells in Mooren ulcer specimens was significantly higher (3.5-fold). However, in the substantia propria, Mooren ulcer specimens revealed significantly increased numbers of CD1alpha+, CD3+, CD4+, CD20+, CD28+, B7-1+, B7-2+, and CD68+ cells. The ratios of CD4+/CD8+ cells and B7-2+/antigen-presenting cells in Mooren ulcer specimens were significantly higher (5-fold). All tested adhesion molecules showed significant up-regulation in the patients' conjunctivas. Mooren ulcer vascular endothelial cells prominently expressed E-selectin, VCAM-1, VLA-4, and ICAM-1 compared with normal conjunctiva. CONCLUSION: The simultaneous presence of multiple types of inflammatory cells, adhesion, and co-stimulatory molecules in Mooren ulcer conjunctiva suggests that their interaction may contribute to a sustained immune activation as at least part of the pathogenic mechanism of this disorder.     

Somatostatin receptor gene expression in human ocular tissues: RT-PCR and immunohistochemical study

PURPOSE: Somatostatin (SST) analogues have been used to treat proliferative diabetic retinopathy, pseudotumor cerebri, thyroid orbitopathy, and cystoid macular edema. There is a paucity of published data in regards to cell-specific distribution of SST receptors (SSTR) in normal human eye tissues. Gene expression for all five known SSTRs in normal human ciliary body/iris, retina, choroid, and cultured retinal pigment epithelial (RPE) cells were studied. METHODS: mRNA was isolated from human ocular tissues (iris/ciliary body, retina, and choroid) dissected from eight pairs of normal eyes (9-62 years) and from RPE cells grown in culture. RT-PCR was done for all five SSTRs in all analyzed tissues. Immunohistochemistry for SSTR1 and SSTR2 was performed on eight pairs of normal human eyes (28-74 years) imbedded in paraffin. RESULTS: SSTR1 to 5 genes are expressed in retina, SSTR1 and SSTR2 genes in cultured RPE cells, and SSTR1, 2, and 4 in ciliary body and choroid. SSTR1 and SSTR2 immunoreactivity (-ir) was observed on a variety of cells within all analyzed tissues including cornea, iris, trabecular meshwork, Schlemm's canal, ciliary processes, ciliary muscle, retina, choroid, cultured RPE cells, and optic nerve. CONCLUSIONS: SSTR genes are widely expressed in normal human eye tissues, with genes for SSTR1 and SSTR2 being the most widely expressed. Genes for all SSTRs are expressed in retina. SSTR1-ir and SSTR2-ir were observed in all analyzed ocular tissues. Detailed knowledge of SSTRs distribution and function in the human eye will result in a better understanding of their role in health and disease.     

Neovascularization in the anterior segment of the rabbit eye by experimental anterior ischemia

PURPOSE: To investigate the effects of anterior ischemia accompanied by neither retinal nor choroidal ischemia on the anterior segment of the eye. METHODS: Both long posterior ciliary arteries in the right eye of 14 rabbits were directly cauterized with an electric coagulator. The eyes were enucleated 1, 2, 4, 7, 9 or 14 days after cauterization, then fixed with 4% paraformaldehyde. Semi-thin sections were studied by light microscopy. Several sections were stained with Griffonia simplicifolia lectin, which bound specifically to mammalian vascular endothelium. Other specimens were examined immunohistochemically for vascular endothelial growth factor (VEGF) protein. The tissue specimens of the first postoperative day were studied for expression of VEGF mRNA by in situ hybridization. RESULTS: Atrophy of the iris and ciliary body was seen after the second postoperative day. Corneal neovascularization appeared after 7 days. Neovascularization on the anterior surface of the iris and in the trabecular meshwork was detected after the ninth postoperative day. The proliferative tissues with newly formed vessels obstructed the iridocorneal angle 14 days after the treatment. There was no histological change in either the retina or choroid. Immunohistochemically, VEGF protein was detected in the epithelial and vascular cells of the iris on the first and fourth postoperative day. Expression of VEGF mRNA was detected in the epithelial cells of the ciliary body on the day following the treatment. CONCLUSIONS: Anterior segment ischemia, when unaccompanied by retinal ischemia, causes neovascularization in the cornea, iris and trabecular tissue.     

Characterization of alpha(1)-adrenoceptor subtypes in the eye

The presence of the alpha(1)-adrenoceptor subtypes in various parts of the pig and rabbit eyes was investigated using [(3)H]-prazosin radioligand binding. The characterization of the subtypes was achieved by performing competition experiments with various subtype selective drugs. In the pig retina, both alpha(1A)- and alpha(1B)-adrenoceptors were detected and the proportion of sites was 70% alpha(1A)- and 30% alpha(1B)-adrenoceptors, respectively. In the pig iris, ciliary body and choroid, which are melanin-rich tissues, the non-specific binding of [(3)H]-prazosin was too high to detect any of the alpha(1)-adrenoceptor subtypes. However, in the albino rabbit iris, ciliary body and retina both alpha(1A)- and alpha(1B)-adrenoceptors were detected. The proportion of sites in the iris was 60 % alpha(1A)- and 40% alpha(1B)-adrenoceptors, respectively. In the ciliary body and rabbit retina the proportion of sites were 70% alpha(1A)- and 30% alpha(1B)-adrenoceptors. Only the alpha(1A)-adrenoceptor subtype was detected in the rabbit choroid.     

Expression and distribution of matrix metalloproteinases and their inhibitors in the human iris and ciliary body

AIM: To determine the expression and distribution of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in the normal human iris and ciliary body. METHODS: Seven postmortem human eyes were fixed with formalin. The iris and ciliary body were dissected out and embedded in paraffin. The expression of MMPs -1, 2, 3, and 9, and TIMPs 1-4 in the iris and ciliary body was determined by a novel immunofluorescence technique and the results graded by masked observers. RESULTS: Positive staining for MMPs and TIMPs was observed in all regions of the anterior uvea, and was more intense in the ciliary body than in the iris. Most MMPs and TIMPs showed similar patterns in their distribution. In the ciliary body, staining was strongest in the epithelium, and was localised to the epithelial cell cytoplasm, except for TIMP-3 which was strongly expressed in the basement membranes. In the iris, staining was most noticeable in the anterior border and anterior epithelial layer. Blood vessels in the stroma of the iris and ciliary body also stained moderately for MMPs and TIMPs. CONCLUSION: Both MMPs and TIMPs are widely expressed in the anterior uvea, with a positive correlation between their expressions. Their differential localisation in the ciliary body suggests they may have a role in maintaining homeostasis in the uveal tract.     

Sympathetic ophthalmia: A clinicopathologic report

A 22-year-old man presented after a pentrating knife injury to the left eye, causing prolapse of the iris, ciliary body, and retinal tissue. The wound was repaired, and antibiotic and steroid therapy was started. At that time, the right eye appeared normal. The patient stopped treatment after discharge and was readmitted 2 weeks later with bilateral panuveitis. His injured eye was enucleated. Pathologic examination confirmed the diagnosis of sympathetic ophthalmia. Presented in part at the XXI International Congress of the International Academy of Pathology, October 20, 1996, Budapest, Hungary.