Molecular And Immunopathology Of Experimental And Clinical Ocular Diseases

通过常规病理、免疫组织化学和分子病理学方法，分析患者标本和动物标本中眼部炎性细胞、变性细胞和肿瘤细胞及其产物的特性和定位。显微解剖等尖端技术的应用，结合了PCR、RT-PCR和RFLP技术，使我们能够对疾病提供更准确的病理诊断(评估)，并指导我们为患者选择合适的治疗方法。我们建立和或应用各种眼部疾病的动物模型，从而了解不同眼病的发病机制。这有助于我们更好地了解疾病的发病机制，并选择更好的针对特定疾病的治疗方法。2007财年，我们继续并完成了以下研究工作： 1.眼淋巴瘤细胞中基因和蛋白的检测：我们继续用显微解剖和聚合酶链式反应证实眼B细胞淋巴瘤的IgH基因重排。与法国的LeHoang教授和她的同事合作，我们发现51例原发性眼内淋巴瘤(PIOL)患者房水中白细胞介素10(IL-10)升高，而108例葡萄膜炎患者未见升高，提示IL-10可能是减少诊断延误的良好筛查试验。明确诊断PIOL需要鉴定眼组织中的PIOL细胞，这可以从玻璃体、视网膜和或视网膜下组织的活检中获得。我们还报告了一例原发性弥漫性大B细胞淋巴瘤合并皮质类固醇激素反应的浆液性视网膜脱离的病例。 2.老年性黄斑变性(AMD)的分子病理学：我们检测到AMD皮损中HtrA-1的单核苷酸多态频率和表达水平均高于正常眼，证实了HtrA-1基因与AMD的相关性研究。 3.眼部疾病的新病理和发病机制：我们与中国的梁博士合作，描述了von Hippel-Lindau(von Hippel-Lindau)相关的视网膜大血管母细胞瘤的手术切除可能对选择VHL疾病的患者有帮助。我们首次在VHL相关的视网膜肿瘤细胞中发现了CXCR4的表达。我们报告了用角膜缘干细胞移植治疗自身免疫性多腺性综合征相关性角膜病变的眼球溶解。我们还报告了婴幼儿肾性胱氨酸病的眼部表现和组织病理学。 4.多种眼部疾病的实验模型：我们发表的一项研究表明，小鼠Ccl2Cx3cr1缺乏会导致类似于人类老年性黄斑变性(AMD)的视网膜病变。这些小鼠在4-5周龄时就出现了人类AMD的临床和病理特征，如玻璃疱疹样变，RPE细胞和Bruchs膜异常，光感受器变性等。脉络膜新生血管也发生在15%的动物身上。Ccl2Cx3cr1缺陷小鼠的视网膜和RPE中A2E含量升高，内质网蛋白含量降低。这种疾病的早期发病和高外显性使这些小鼠成为人类AMD的易感模型。 在与NEI的Caspi、Gery、Hooks和Nussenblatt博士的合作下，已经发现并发表了几种新的眼部炎症模式和机制，并在他们的年度报告中进行了描述。

The identity and topographic location of ocular inflammatory, degenerative, and tumor cells and their products in patient specimens and animal samples are analyzed by routine pathology, immunohistochemistry, and molecular pathology. The application of cutting-edge technology such as microdissection combined with PCR, RT-PCR, and RFLP techniques, allows us to provide more accurate pathological diagnosis (assessment) of the disease, and guiding us in selecting an appropriate therapy for the patient. We generate andor apply various animal models with ocular disorders, so we can learn the pathogenesis of different ocular diseases. This helps us to better understand disease pathogenesis and to select better therapies targeting specific diseases. In FY2007, we continued and accomplished the following in our research: 1. Detection of Genes and Proteins in Ocular Lymphoma Cells: We continue to confirm IgH gene rearrangements of ocular B-cell lymphoma using microdissection and PCR. In collaboration with Prof. LeHoang and her colleagues of France, we found elevation of aqueous interleukin (IL)-10 in 51 patients with primary intraocular lymphoma (PIOL) but not in the 108 uveitic patients, suggesting that IL-10 may be a good screening test to reduce delay in diagnosis. The defined diagnosis of PIOL requires the identification of PIOL cells in ocular tissues, which can be obtained from biopsies of the vitreous, retina andor subretinal tissues. We also reported a case of primary diffuse large B-cell lymphoma of the spleen with coincident serous retinal detachment responsive to corticosteroids. 2. Molecular Pathology of Age-Related Macular Degeneration (AMD): We detected higher rates of single nucleotide polymorphisms and higher expression of HtrA-1 in the AMD lesions compared to normal eyes, confirming the association studies of HtrA-1 gene and AMD. 3. New Pathology and Pathogenesis of Ocular Diseases: In collaboration with Dr. Liang of China, we described surgical resection of von Hippel-Lindau (VHL)-associated large retinal hemangioblastomas might be useful for therapy in select patients with VHL disease. We demonstrated CXCR4 expression for the first time in VHL-associated retinal tumor cells. We reported ocular resolution of autoimmune polyglandular syndrome associated keratopathy with keratolimbal stem cell transplantation. We also presented ophthalmic manifestations and histopathology of infantile nephropathic cystinosis. 4. Experimental Models for Various Ocular Diseases: We published a study showing that murine Ccl2Cx3cr1 deficiency results in retinal lesions mimicking human age-related macular degeneration (AMD). These mice present with clinical and pathological features of human AMD, such as drusen-like lesions, abnormal RPE cells and Bruchs membrane, and photorecptor degeneration as early as 4-5 weeks of age. Choroidal neovascularization also occurs in 15% of the animals. Elevation of A2E and decreased amounts of endoplasmic reticulum protein in the retina and RPE were also measured in the Ccl2Cx3cr1 deficient mice. The early onset and high penetrance of the disease make these mice an attarctive model for human AMD. In collaboration with Drs. Caspi, Gery, Hooks, and Nussenblatt of the NEI, several new modes and mechanisms of ocular inflammation have been discovered and published, which are described in their annual reports.