COMPLEMENT REGULATION IN FETOMATERNAL TOLERANCE

胎儿母体耐受性的补体调节

• 批准号: 2827237
• 负责人: HECTOR D MOLINA
• 金额: $ 23.21万
• 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/2827237
• 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(RCA)蛋白调节蛋白使补体(C)级联失活，似乎保护组织免受C介导的损伤。然而，目前尚不清楚RCA功能的异常如何影响体内的免疫系统。为了研究它们在健康和疾病中的作用，通过基因打靶产生了CRY/P65基因缺陷的小鼠。Crry-/-小鼠无法存活到胚胎阶段。对胎儿胎盘组织的检查表明，母胎界面C的激活是导致胎儿丢失的原因。饲养Crry+/-动物到C3-/-(C3-/-Crry-/-)小鼠拯救了这种致命的表型。在这个项目中，将通过1)研究Crry/p65和其他小鼠RCA蛋白在胚胎发育的不同阶段在Crry+/+、Crry+/-和Crry-/-小鼠胎盘组织中的表达；2)研究Crry/p65的异常表达和C沉积是如何影响胎儿-母体相互作用的，从而进一步表征Crry/p65在胚胎发育和胎儿胎盘耐受中的作用。将探讨胎儿胎盘单位内的形态异常，以及组织对炎症、细胞溶解和C沉积的其他有害后果的易感性。将尝试使用可溶性形式的Crry/p65或表达Crry/p65的转基因小鼠来挽救胚胎的致死性。此外，利用免疫组织化学方法，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。采用不同的体内炎症模型，通过监测机体对免疫复合物攻击的反应来分析炎症反应和免疫复合体介导的损伤程度。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，在这些小鼠中，C3缺乏已通过野生型骨髓移植部分纠正。这些实验将有助于理解补体和补体调节蛋白在健康和疾病中的关键作用。此外，它还将使我们能够分析C和RCA蛋白在母胎耐受和生殖系统中的作用。