STUDIES OF SOLUBLE IMMUNOREGULATORS IN MURINE LUPUS

鼠狼疮可溶性免疫调节剂的研究

• 批准号: 3454268
• 负责人: HARUMI JYONOUCHI JYONOUCHI
• 金额: $ 9.8万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3454268
• 关键词: B lymphocyte; affinity chromatography; autoimmunity; chimeric proteins; cytokine; immunoregulation; laboratory mouse; leukopoiesis; molecular cloning; monoclonal antibody; northern blottings; nucleic acid sequence; recombinant proteins; systemic lupus erythematosus

NZB mice are the first experimental animal model of human lupus. In previous studies we have demonstrated a characteristic hyperactive B lymphopoiesis in the early life of the NZB mice. However, this is a temporary phenomenon and declines rapidly prior to the manifestation of autoimmune phenomena and the development of autoimmune disease. Recently we have identified humoral factors which can enhance sIg-B acquisition and colony formation by sIg B precursor cells from normal strains of mice in vitro. They were first identified in the serum of very young NZB mice when they were active in B lymphopoiesis and thus termed NZB serum factors (NZB-SF). The NZB-SF activity in serum decreases with age in parallel to the decline of once hyperactive B of NZB-SF and designated as the 60 kd NZB-SF. Serological studies and analyses of biological functions of this molecule indicate that this is distinguishable from well defined cytokines. We hypothesized that in NZB mice the overproduction of NZB-SF may be, at least in part responsible for the premature hyperactive B lymphopoiesis and the dysregulation autoimmunity in later life. The long term objective of this proposal is to test this hypothesis and elucidate the roles of the 60 kd NZB-SF in relation to autoimmunity. The immediate goals are to characterize the 60 kd NZB-SF further and to define its biological functions. For that purpose, we propose 4 specific aims. They are 1) molecular characterization of 60 kd NZB-SF by cloning genes and analyzing DNA sequence and recombinant protein products of cloned cDNA coding for the 60 kd NZB-SF, 2) identification of cellular origin of the 60 kd NZB-SF utilizing ELISA, bioassays, immunocytochemical analysis, northern blot analysis, and possibly in situ hybridization technique, 3) characterization of NZB-SF present in NZB serum using immunoaffinity membrane chromatography, 4) analyses of effects of 60 kd NZB-SF on B lymphopoiesis in vivo by administration of NZB-SF to mice of non-autoimmune strains or alternatively by administration of mAb against NZB-SF to young NZB mice.

NZB小鼠是人类狼疮的第一个实验动物模型。 在 以前的研究表明，一个典型的多动B 在NZB小鼠的早期生命中的淋巴细胞生成。 但这是一 暂时的现象，并迅速下降之前的表现， 自身免疫现象和自身免疫疾病的发展。 最近，我们已经确定了体液因子，可以增强sIg-B 正常人sIg B前体细胞的获得和集落形成 品系小鼠体外培养。 它们最初是在一种 年轻的NZ B小鼠，当它们在B淋巴细胞生成中活跃时，因此称为NZ B 血清因子（NZB SF）。 血清NZB SF活性随年龄增长而降低， 与NZB-SF的一度过度活跃的B的下降平行，并指定为 60 kd NZB-SF。 血清学研究和生物学功能分析 表明这与定义明确的 细胞因子 我们假设，在NZB小鼠中，NZB-SF的过量产生可能是， 至少部分负责过早的过度活跃的B淋巴细胞生成， 自身免疫功能失调 的长期目标 这一建议是为了验证这一假设，并阐明60 kd NZB-SF与自身免疫相关。 近期目标是 进一步表征60 kd NZB-SF并确定其生物学特性， 功能协调发展的 为此，我们提出了四个具体目标。 它们是1）分子 通过克隆基因和分析DNA鉴定60 kd NZB-SF 克隆的编码60个氨基酸的cDNA序列和重组蛋白产物 kd NZB-SF，2）60 kd NZB-SF的细胞来源的鉴定 利用ELISA、生物测定、免疫细胞化学分析、北方印迹 分析和可能的原位杂交技术，3）表征 使用免疫亲和膜检测NZB血清中存在的NZB-SF 60 kd NZB SF对B淋巴细胞增殖的影响 通过向非自身免疫品系的小鼠施用NZB-SF体内，或 或者通过向年轻的NZB小鼠施用抗NZB-SF的mAb。