GENETIC REGULATION OF THE HEPATIC ACUTE PHASE RESPONSE

肝脏急性期反应的基因调控

• 批准号: 6329320
• 负责人: HEINZ BAUMANN
• 金额: $ 24.87万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6329320
• 关键词: Adenoviridae; acute phase protein; animal genetic material tag; bactericidal immunity; gene induction /repression; gene mutation; genetic promoter element; genetically modified animals; haptoglobins; human tissue; immunogenetics; inflammation; laboratory mouse; liver; protein structure function; tissue /cell culture; transcription factor; transfection /expression vector

DESCRIPTION: Tissue damage and infection initiate a local inflammatory reaction that is designed to mobilize the immediate defense mechanisms of the organism, constituting the acute phase (AP) response and innate immune responses. Induction of hepatic acute phase plasma protein production is one of the systemic effects elicited by all inflammatory processes. Haptoglobin (Hp), the pronciple hemoglobin-binding protein in the plasma is one of the major APPs in mammals. Hp moderates the functions of inflammatory cells, and contributes to a bacteriostatic milieu. The project's goal is to define, in the mouse model, the roles of AP induced Hp controlling inflammatory responses. The specific aims are: 1) To identify the mechanism of Hp gene induction in cell culture. Three types of experiments are proposed all of which follow standard procedures employing transient transfection into HepG2 cells. In the first, sequence elements and factors which bind to a 115 bp region of the Hp promoter will be further dissected. In the second, exon shuffling between Stat1 and Stat3 is proposed to define functional domains of the latter in this system. In the third, sequence element identification is proposed outside the proximal promoter region. 2) To establish Hp-deficient mice. Inbred and congenic Hp null mice will be established. The response of Hp +/- and Hp -/- animals to representative inflammatory challenges, including tissue damage, endotoxin and infections will be defined by the pattern of acute phase protein genes induced in the liver and changes in composition of acute phase proteins and leukocytes in the blood. 3) To verify the specific action of Hp in these null animals and cell culture systems. Differences in response pattern may suggest potential functions of Hp which will be verified in Hp null animals by Hp administration or adenovirally re-introduced hepatic Hp production. The cellular action of Hp on differentiated functions associated with inflammatory cells will be determined in tissue culture systems. 4) To determine the consequence of a deleted acute phase induction of Hp expression. Mice null of the IL-6 response element in the Hp promoter will be generated by homologous recombination and targeted mutatgenesis and subjected to acute phase stimulants to assess the contribution of these sequence to Hp gene regulation.

描述：组织损伤和感染引发局部炎症 反应，旨在动员直接防御机制， 生物体，构成急性期（AP）反应和先天免疫 应答 肝急性期血浆蛋白产生的诱导是 是所有炎症过程引起的全身效应之一。 结合珠蛋白（Hp）是血浆中主要的血红蛋白结合蛋白， 是哺乳动物的主要APP之一。 Hp调节了 炎症细胞，并有助于抑菌环境。 的 项目的目标是在小鼠模型中确定AP诱导的Hp的作用， 控制炎症反应。 具体目标是：1）识别 Hp基因在细胞培养中的诱导机制。 三种类型的 所有这些实验都遵循标准程序， 瞬时转染HepG 2细胞。 首先，序列元素 结合Hp启动子的115 bp区域的因子将被进一步 解剖。 在第二种情况下，Stat 1和Stat 3之间的外显子改组是 建议在该系统中定义后者的功能域。 在 第三，提出了近端外的序列元件识别， 启动子区 2)建立幽门螺杆菌缺陷小鼠模型。 近交系和同源系Hp 将建立无效小鼠。 Hp +/-和Hp -/-动物对 代表性炎症挑战，包括组织损伤、内毒素 感染将由急性期蛋白基因的模式来定义 在肝脏中诱导和急性时相蛋白组成的变化， 血液中的白细胞 3)为了验证Hp在这些疾病中的特异性作用， 空动物和细胞培养系统。 响应模式的差异可能 提示Hp的潜在功能，其将在Hp缺失动物中验证 通过Hp施用或腺病毒重新引入肝Hp产生。 幽门螺杆菌对分化功能的细胞作用 将在组织培养系统中测定炎性细胞。 4)到 确定Hp急性期诱导缺失的后果 表情 Hp启动子中IL-6应答元件缺失的小鼠将 通过同源重组和靶向突变产生， 进行急性期兴奋剂，以评估这些贡献 序列与Hp基因调控的关系。