Generation of complement C9 conditional knockout mice and anti-C9 mAbs

描述（由申请人提供）：补体是先天免疫系统的重要组成部分，并且可以说最为人所知的是其通过称为膜攻击复合物（MAC）的大分子结构裂解细菌、包膜病毒和真核细胞的能力。MAC由几种补体蛋白（C5 b、C6、C7、C8和C9）组成，但C9是MAC的成孔结构所需的关键蛋白。由于补体系统在炎症条件下对自身和非自身的区分较差，因此补体的不适当激活和随后的MAC介导的自身组织破坏是自身免疫性疾病的常见特征。在这种情况下，将C9与细胞死亡联系起来的证据通常是间接的，主要基于死后组织样本中C9的免疫组织化学染色。然而，在某些疾病环境中，例如脱髓鞘疾病，对C9和MAC作为致病机制的重要组分的需求之间存在明显的差异。这就提出了关于C9在脱髓鞘疾病和其他自身免疫性疾病中的重要性的问题，其中补体介导的机制被认为是疾病病理学的核心。此外，关于C9生物学还有许多未解答的问题。例如，C9是否有助于正常发育？C9基因表达的调控机制是什么？C9与原核和真核细胞膜相互作用产生的信号事件是什么？令人惊讶的是，没有专门针对老鼠的工具来解决这些问题。我们建议产生C9特异性单克隆和多克隆抗体和条件C9缺陷小鼠，以提供急需的工具来直接评估C9在自身免疫性和炎症性疾病中的作用。公共卫生相关性：由于缺乏合适的试剂，补体成分C9在自身免疫性疾病和感染性疾病中的作用仍然很少探索。本申请中的研究旨在产生C9条件性敲除小鼠和抗C9单克隆抗体。

DESCRIPTION (provided by applicant): Complement is important part of the innate immune system and is arguably best known for its ability to lyse bacteria, enveloped viruses and eukaryotic cells through a macro- molecular structure known as the membrane attack complex (MAC). The MAC is composed of several complement proteins (C5b, C6, C7, C8 and C9), but C9 is the critical protein required for the pore-forming structure of the MAC. Because the complement system discriminates poorly between self and non-self under inflammatory conditions, inappropriate activation of complement and subsequent MAC-mediated destruction of self-tissues is a common feature of autoimmune disease. The evidence linking C9 to cell death in this setting is frequently circumstantial, based largely on immunohistochemical staining for C9 in postmortem tissue samples. However in some disease settings, such as demyelinating disease, there are clear discrepancies between the need for C9 and the MAC as important components of the pathogenic mechanism. This raises questions regarding how critical C9 is in demyelinating disease and in other autoimmune diseases where complement-mediated mechanisms are considered central to disease pathology. In addition, there are many unanswered questions regarding C9 biology. For example, does C9 contribute to normal development? What are the regulatory mechanisms for C9 gene expression? What are the C9-mediated signaling events generated on interaction with prokaryotic and eukaryotic membranes? Surprisingly, there are no murine-specific tools to address these questions. We propose to generate C9-specific monoclonal and polyclonal antibodies and conditional C9-deficient mice to provide much needed tools to directly assess of the role of C9 in autoimmune and inflammatory diseases. PUBLIC HEALTH RELEVANCE: The role of the complement component C9 in autoimmune and infectious disease remains poorly explored due to the lack of appropriate reagents. Studies in this application are designed to generate C9 conditional knockout mice and anti-C9 monoclonal antibodies.