Cytokine mechanisms in systemic lupus erythematosus

系统性红斑狼疮的细胞因子机制

• 批准号: 7073434
• 负责人: CHAIM O. JACOB
• 金额: $ 39.78万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7073434
• 关键词: T lymphocyte; autoantibody; bioinformatics; gene expression profiling; gene mutation; genetically modified animals; growth factor receptors; interferon gamma; interleukin 7; laboratory mouse; microarray technology; molecular pathology; monoclonal antibody; phenotype; protein isoforms; renal glomerulus; systemic lupus erythematosus; transcription factor

DESCRIPTION (provided by applicant): The pathogenesis of systemic lupus erythematosus (SLE), a prevalent human systemic autoimmune disease, is still unknown. Thus, efforts aimed at understanding disease mechanisms are highly relevant. Because studying humans with SLE is difficult for multiple reasons, we have developed novel genetically lupus-prone NZBxNZW derived congenic NZM lines that are either Stat-4 or Stat-6 deficient. The Stat deficient mice offer unique tools to achieve a better understanding of the role that autoAbs, and cytokines play in the pathogenesis of the disease and may allow dissection of the mechanisms that mediate renal damage in SLE. Backed up by substantial preliminary data, we propose to address the following questions: (1) Is the kidney disease in Stat4 deficient lupus mice dependent or independent of autoAbs? To this end we have produced B-cell deficient NZM2328 mutant mice into which we will restore B cells in the absence of circulating, soluble autoAbs and then cross these mice with Stat4 KO. (2) Is IFN-gamma necessary for the development of autoAbs and target organ disease in Star-deficient NZM mice? This specific aim will test the hypothesis that the absence of IFNgamma may result in early initiation of disease through a failure of regulatory mechanisms in the Stat4-/- mice. To accomplish this we will test directly and systematically the role of IFNgamma at different stages of lupus-like disease development in Stat deficient NZM mice using two different approaches: a) in vivo treatment protocols using IFNgamma and anti-IFNgamma reagents for defined periods of time, and b) a genetic approach in which a conditional expression system for IFNgamma in a tissue-and time-specific manner will be developed. (3) Can we reconstitute the Stat deficient phenotypes in immunodeficient NZM mice? To accomplish this, we will generate NZM2328 completely immunodeficient by introducing a loss of function mutation in both the IL-7R alpha and c-kit receptors that allows hematopoietic reconstitution with any donor strain without requiring administration of pre-transplant conditioning. (4) What are the direct effects of the two Stat4 isoforms on the immune system and on the target organ in NZM2328 lupus mice? For this purpose we will restore Stat4 in T lymphocytes of Stat4 deficient NZM2328 mice by crossing them with Stat4- alpha and Stat4-beta transgenic mice and will study differentially expressed genes in lymphocytes and glomeruli of these lupus mice using micro-array technology. These studies should lead to better understanding of a number of characteristics of SLE and may lead to identification of new targets for therapy.

描述（由申请人提供）：系统性红斑狼疮（SLE）是一种常见的人类系统性自身免疫性疾病，其发病机制尚不清楚。因此，旨在了解疾病机制的努力是高度相关的。由于多种原因，研究SLE患者很困难，我们开发了一种新的遗传上易患狼疮的NZBxNZW衍生的基因NZM系，这些系要么缺乏Stat-4，要么缺乏Stat-6。Stat缺陷小鼠提供了独特的工具，以更好地了解自身抗体和细胞因子在疾病发病机制中的作用，并可能允许解剖介导SLE肾损害的机制。在大量初步数据的支持下，我们提出解决以下问题：(1)Stat4缺陷狼疮小鼠的肾脏疾病是依赖还是独立于自身抗体？为此，我们生产了B细胞缺陷的NZM2328突变小鼠，我们将在缺乏循环可溶性自身抗体的情况下恢复B细胞，然后将这些小鼠与Stat4 KO杂交。(2) ifn - γ在缺星NZM小鼠自身抗体和靶器官疾病的发生中是否必需？这一特定目的将验证这样一种假设，即在Stat4-/-小鼠中，缺乏IFNgamma可能通过调节机制的失败导致疾病的早期开始。为了实现这一目标，我们将使用两种不同的方法直接和系统地测试IFNgamma在Stat缺乏的NZM小鼠狼疮样疾病发展的不同阶段的作用：a)在确定的时间内使用IFNgamma和抗IFNgamma试剂的体内治疗方案，b)一种遗传方法，其中IFNgamma的条件表达系统将以组织和时间特异性的方式开发。(3)能否重建免疫缺陷的NZM小鼠Stat缺陷表型？为了实现这一目标，我们将通过在IL-7R α和c-kit受体中引入功能缺失突变来产生完全免疫缺陷的NZM2328，这种突变允许在不需要移植前调节的情况下与任何供体株进行造血重建。(4)两种Stat4亚型对NZM2328狼疮小鼠免疫系统和靶器官的直接影响是什么？为此，我们将通过将缺乏Stat4的NZM2328小鼠与Stat4- α和Stat4- β转基因小鼠杂交，恢复其T淋巴细胞中的Stat4，并使用微阵列技术研究这些狼疮小鼠淋巴细胞和肾小球中的差异表达基因。这些研究将有助于更好地了解SLE的许多特征，并可能导致确定新的治疗靶点。