COMPLEMENT REGULATION IN FETOMATERNAL TOLERANCE

小鼠Crry/p65蛋白和其他补体激活调节因子（RCA）蛋白灭活补体(C)级联，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缺乏Crry/p65的小鼠。cry -/-小鼠不能在胚胎期存活。对胎胎盘组织的检查表明，C在胎母界面的激活是导致胎儿丢失的原因。将Crry+/-动物与C3-/- (C3-/-Crry-/-)小鼠杂交可以挽救这种致命的表型。本项目将通过1)研究Crry/p65及其他小鼠RCA蛋白在不同胚胎发育阶段的Crry+/+、Crry+/-和Crry-/-小鼠胎胎盘组织中的表达，进一步表征Crry/p65在胚胎发育和胎胎盘耐受中的作用；2)研究异常的Crry/p65表达和C沉积如何影响胎母相互作用。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被探讨。将尝试使用可溶性的Crry/p65或表达Crry/p65的转基因小鼠来挽救胚胎致死。此外，利用免疫组织化学，C被激活的机制将被确定。最后，将监测炎症或异常免疫行为的存在。通过使用不同的体内炎症模型监测对免疫复合物挑战的反应，将研究炎症反应和免疫复合物介导的损伤程度。在这些实验中，将Crry+/+小鼠与Crry+/-小鼠进行比较。此外，我们将比较C3-/-Crry -/-和C3-/-Crry+/+小鼠，其中C3缺乏已通过野生型骨髓移植得到部分纠正。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。这也将使我们能够分析C和RCA蛋白在母婴耐受和生殖系统中的作用。

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.