COMPLEMENT REGULATION IN FETOMATERNAL TOLERANCE

胎儿母体耐受性的补体调节

• 批准号: 6511141
• 负责人: HECTOR D MOLINA
• 金额: $ 25.36万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-15 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6511141
• 关键词: complement; complement pathway; developmental immunology; genetically modified animals; immune complex; immune tolerance /unresponsiveness; inflammation; laboratory mouse; mammalian embryology; placenta; polymerase chain reaction; pregnancy immunology; tissue /cell culture

The murine Crry/p65 protein and other Regulators of Complement Activation (RCA) proteins inactivate the complement (C) cascade and appear to protect tissues from C-mediated damage. Nevertheless, it is not known how abnormalities in the function of the RCA affect the immune system in vivo. To investigate their role in health and disease, mice deficient in Crry/p65 have been generated by gene targeting. Crry-/-mice do not survive the embryonal stage. Examination of fetoplacental tissues suggests that activation of C at the fetomaternal interface is causing the fetal loss. Breeding Crry+/- animals to C3-/- (C3-/-Crry-/-) mice rescue this lethal phenotype. In this project further characterization of the role of Crry/p65 in embryonic development and in fetoplacental tolerance will be analyzed by 1) studying the expression of Crry/p65 and other mouse RCA proteins in fetoplacental tissues of Crry+/+, Crry+/-, and Crry-/- mice during different stages of embryonic development; 2) investigating how abnormal Crry/p65 expression and C deposition is affecting the fetomaternal interact. Morphological abnormalities within the fetoplacental unit, and the susceptibility of the tissue to inflammation, cell lysis, and other deleterious consequences of C-deposition will be explored. Rescuing of embryonic lethality using soluble forms of Crry/p65 or transgenic mice expressing Crry/p65 will be attempted. Furthermore, using immunohistochemistry, the mechanism by which C is activated will be determined. Finally, the presence of inflammation or aberrant immune behavior will be monitored. Analysis of the inflammatory reaction and the degree of immune complex mediated injury will be studied by monitoring the response to immune complex challenge using different in vivo models of inflammation. In these experiments, Crry+/+ mice will be compared to Crry+/- mice. In addition, we will compare C3-/- Crry-/- and C3-/-Crry+/+ mice in which the C3 deficiency has been partially corrected by wild type bone marrow transfer. These experiment will help understand the critical role of complement and complement regulatory proteins in health and disease. Also it will also enable us to analyze the role of C and the RCA proteins in fetomaternal tolerance and in the reproductive system.

小鼠 Crry/p65 蛋白和其他补体激活调节剂 (RCA) 蛋白可使补体 (C) 级联失活，并似乎可以保护组织免受 C 介导的损伤。然而，目前尚不清楚 RCA 功能异常如何影响体内免疫系统。 为了研究它们在健康和疾病中的作用，通过基因靶向产生了 Crry/p65 缺陷的小鼠。 哭泣的小鼠无法在胚胎阶段存活。 对胎儿胎盘组织的检查表明，胎儿母体界面处 C 的激活导致胎儿流产。 将 Crry+/- 动物繁殖为 C3-/- (C3-/-Crry-/-) 小鼠可挽救这种致命表型。 在本项目中，将通过以下方式进一步分析 Crry/p65 在胚胎发育和胎儿胎盘耐受中的作用：1) 研究 Crry+/+、Crry+/- 和 Crry-/- 小鼠在胚胎发育不同阶段的胎儿胎盘组织中 Crry/p65 和其他小鼠 RCA 蛋白的表达； 2) 研究异常的 Crry/p65 表达和 C 沉积如何影响胎儿母体相互作用。 将探讨胎儿胎盘单位内的形态异常，以及组织对炎症、细胞裂解和碳沉积的其他有害后果的敏感性。将尝试使用可溶形式的 Crry/p65 或表达 Crry/p65 的转基因小鼠来挽救胚胎致死性。此外，使用免疫组织化学，将确定 C 被激活的机制。 最后，将监测炎症或异常免疫行为的存在。将通过使用不同的体内炎症模型监测对免疫复合物攻击的反应来研究炎症反应和免疫复合物介导的损伤程度的分析。 在这些实验中，Crry+/+ 小鼠将与 Crry+/- 小鼠进行比较。 此外，我们将比较 C3-/- Crry-/- 和 C3-/-Crry+/+ 小鼠，其中 C3 缺陷已通过野生型骨髓移植部分纠正。 这些实验将有助于了解补体和补体调节蛋白在健康和疾病中的关键作用。 它还将使我们能够分析 C 和 RCA 蛋白在胎儿母体耐受性和生殖系统中的作用。