IMMUNOBIOLOGY OF TAMARIN COLITIS

狨猴结肠炎的免疫生物学

• 批准号: 3243425
• 负责人: James K. Roche
• 金额: $ 13.34万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3243425
• 关键词: Ceboidea; antibody formation; antigen antibody reaction; autoantigens; autoimmune disorder; cell mediated lymphocytolysis test; cellular immunity; chromium; colitis; cytotoxic T lymphocyte; disease /disorder model; enzyme linked immunosorbent assay; epitope mapping; flow cytometry; gastrointestinal epithelium; glycoproteins; helper T lymphocyte; immunoregulation; laboratory rat; leukocyte activation /transformation; radionuclides; radiotracer; tissue /cell culture; tritium; western blottings

For many chronic inflammatory diseases, antigen-specific immune responses are causative of inflammation and/or dysfunction of a target organ (autoimmune thyroiditis, with of inflammation and/or dysfunction of a target organ (autoimmune thyroiditis, with thyroglobulin; myasthenia gravis, with acetylcholine receptor; post-vaccination encephalomyelitis with myelin basic protein). Given that most of these human disorders are spontaneous and thus unpredictable in onset, animal models have been crucial in working out their basic pathophysiology. For the chronic inflammatory bowel diseases (ulcerative colitis, Crohn's disease) affecting up to 30/100,000 population, epithelium-specific immune responses have been described after disease onset in research studies for over two decades, but there has been no animal model for testing their role in induction or maintenance of tissue injury. The New World monkey, S. oedipus (or cotton-top tamarin), as a recently described primate with a spontaneous chronic colitis, is of considerable interest because (a) animals can be studied prior to lesion onset and serial observations can be made on mucosal histology as well as on immune phenomena; (b) clinically, they resemble the human disease in several ways, including response to therapy (see Background); and (c) antigen-specific immune phenomena, parallel to those in human chronic ulcerative colitis, have been specific immune phenomena, parallel to those in human chronic ulcerative colitis, have been recently described. These include serum immunoglobulin specific for epithelial glycoproteins (designated epithelial cell-associated components, ECAC), up to 18 mug Ig/ml; determinants on their intestinal epithelium reactive with ECAC-specific monoclonal antibodies by immunofluorescence; cytotoxicity against ECAC-labeled targets (using tamarin immunoglobulin and normal mononuclear cells); and Ig binding for one fraction (P1) of macromolecules in ECAC. The long-term objectives of this proposal is to define the role of anti-epithelial glycoprotein- specific immune responses for gut tissue injury occurring in this animal model of spontaneous colitis. Specific aims, then, will be to determine: (1) in what way anti-epithelialglycoprotein immune responses in the CTT are similar to responses in classical autoimmune disease; (2) which macromolecules on human intestinal epithelial cells, derived from stable and established cell lines, bind immunoglobulin from tamarins with spontaneous colitis; and (3) what are the characteristics of T lymphocyte reactivity to intestinal epithelial glycoproteins in the tamarin, particularly with respect to location (gut-associated mesenteric LN versus popliteal LN or spleen); the major T lymphocyte subset involved (CD4 versus CD8); the precise macromolecular in ECAC recognized by CTT T-lymphocytes; and ability to carry out ECAC-specific lysis of labeled target cells. The work proposed is highly important in that (a) a sizeable proportion of the experiments performed will be with immunoglobulin derived from a described model of human disease, and (b) better therapy for these disorders in humans will likely be developed when the role of epithelium-specific immune responses, long described in these diseases, is better understood. Experimental methodology will include: techniques for quantitating ECAC- specific antibody (ELISA; Chromium release assay, immunoblotting); techniques for epitope mapping radiolabeling, radioimmune inhibition binding); sterile cell culture techniques for CaCo-2 and T84 cells as well as for the cytotoxicity assay; and 3H-thymidine proliferation assay to define antigen-specific T lymphocytes after enrichment/depletion of T-cell subsets by monoclonal antibody panning as monitored by FACS.

对于许多慢性炎症性疾病，抗原特异性免疫反应 是靶器官发炎和/或功能障碍的原因 (自身免疫性甲状腺炎，伴有炎症和/或功能障碍 靶器官(自身免疫性甲状腺炎，伴甲状腺球蛋白；肌无力 Gravis，有乙酰胆碱受体；接种后脑脊髓炎 具有髓鞘碱性蛋白)。鉴于这些人类疾病中的大多数是 自发的，因此在发病时不可预测的动物模型 对研究他们的基本病理生理学至关重要。对于慢性病患者 炎症性肠病(溃疡性结肠炎、克罗恩病)影响 高达30/100,000人口，上皮特异性免疫反应 在两年多的研究研究中描述了疾病发作后的情况 几十年，但还没有动物模型来测试它们在 引起或维持组织损伤。新大陆猴S。 俄狄浦斯(或棉顶金丝雀)，作为最近被描述的一种灵长类动物 自发性慢性结肠炎，有相当大的兴趣，因为(A) 可以在病变发生之前对动物进行研究，并可以进行连续观察 在粘膜组织学和免疫现象方面；(B)临床上， 它们在几个方面与人类疾病相似，包括对 治疗(见背景)；和(C)抗原特异性免疫现象， 与人类慢性溃疡性结肠炎相似，具有特异性 免疫现象，与人类慢性溃疡性结肠炎的免疫现象相似， 最近都被描述过。其中包括血清免疫球蛋白特异性 上皮糖蛋白(指定为上皮细胞相关 成分，ECAC)，高达18毫克Ig/毫升；对其肠道的决定因素 与ECAC特异性单抗反应的上皮细胞 免疫荧光；对ECAC标记靶标的细胞毒性(使用 他莫林免疫球蛋白和正常单核细胞)；和免疫球蛋白结合 ECAC中大分子的一种组分(P1)。的长期目标 这项提议是为了定义抗上皮糖蛋白的作用- 对该动物肠道组织损伤的特异性免疫反应 自发性结肠炎模型。因此，具体目标将是确定： (1)CTT中的抗上皮糖蛋白免疫反应是以什么方式进行的 类似于经典自身免疫性疾病的反应；(2) 人肠上皮细胞上的大分子，来源于稳定 并建立了细胞系，将罗望子蛋白中的免疫球蛋白与 自发性结肠炎；和(3)T淋巴细胞的特征是什么 罗望子对肠道上皮糖蛋白的反应性， 尤其是关于位置(肠道相关的肠系膜LN与 主要受累T淋巴细胞亚群(CD4VS CD8)；CTT T淋巴细胞识别的ECAC中的精确大分子； 以及对标记的靶细胞进行ECAC特异性裂解的能力。这个 建议的工作非常重要，因为(A)相当大比例的 所进行的实验将使用从所描述的 人类疾病的模型，以及(B)更好地治疗这些疾病 人类很有可能在发育过程中发挥上皮特异性免疫作用 在这些疾病中长期描述的反应更容易被理解。 实验方法学将包括：ECAC定量技术-- 特异性抗体(ELISA；铬释放试验、免疫印迹)； 表位定位技术放射性标记、放射免疫抑制 Caco-2和T84细胞的无菌细胞培养技术 至于细胞毒性试验，以及~3H-胸腺嘧啶核苷增殖试验 T细胞浓缩/耗尽后抗原特异性T淋巴细胞的鉴定 用流式细胞仪监测的单抗淘洗亚群。