Isolation of Autoantibodies to Apoptotic Cells

凋亡细胞自身抗体的分离

• 批准号: 6730607
• 负责人: Marko Z Radic
• 金额: $ 7.15万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-15 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6730607
• 关键词: B lymphocyte; antibody formation; antinuclear autoantibody; apoptosis; autoimmune disorder; cellular pathology; confocal scanning microscopy; flow cytometry; gene mutation; hybridomas; immune tolerance /unresponsiveness; immunoglobulin genes; immunologic receptors; laboratory mouse; phagocytic dysfunction

DESCRIPTION( provided by the applicant): Compelling evidence links the initiation and progression of Systemic Lupus Erythematosus (SLE) and Anti-Phospholipid Syndrome (APS) to aberrant immune responses to apoptotic cells. However, the circumstances that lead to the production of anti-nuclear autoantibodies, the hallmark of SLE, and of autoantibodies to complexes between serum proteins and membrane phospholipids, the serologicalI markers for APS, remain poorly understood. B cells with Ig receptors for apoptotic cells may be in a position to gain access to relevant autoantigens. Following positive selection by autoreactive T cells, such B cells may be induced to secrete autoantibodies to apoptotic cells (ATAC), and disrupt normal clearance mechanisms, thus perpetuating and even expanding the autoimmune response. Alternatively, ATAC may arise secondary to defects in clearance, or an overabundance of apoptotic cell remnants. To distinguish between these possibilities, more detailed information on the genetics, regulation, and function of ATAC is needed. This proposal aims to derive ATAC from mice with defects in phagocytosis of apoptotic cells. The specific aims are: 1) Test sera of Mer, Axl, Tyro3 triply nullizygous mice for the expression of ATAC, and construct hybridomas from positive mice; 2) Screen hybridomas for ATAC specificity by flow cytometry and confocal microscopy and survey their relationship to other autoreactive hybridomas from the same fusions; 3) Determine the genetic basis for ATAC specificity and compare the contributions of immunoglobulin heavy and light chain V gene use, junctional diversity, and somatic mutations to the recognition of apoptotic cells. Future studies will explore the contribution of ATAC-bearing B cells to autoimmune responses against nuclear and cell surface antigens, the effect of soluble ATAC on the clearance of apoptotic cells by macrophage, and the regulation of ATAC-bearing B cells. The results of these studies will allow a fuller assessment of the role of apoptotic cells in the induction of B cell tolerance or autoimmunity.

描述（由申请人提供）：令人信服的证据将系统性红斑狼疮 (SLE) 和抗磷脂综合征 (APS) 的发生和进展与凋亡细胞的异常免疫反应联系起来。然而，导致抗核自身抗体（SLE 的标志）以及血清蛋白和膜磷脂之间的复合物（APS 的血清学标记物）自身抗体产生的情况仍然知之甚少。具有凋亡细胞 Ig 受体的 B 细胞可能能够接触到相关的自身抗原。经过自身反应性 T 细胞的阳性选择后，此类 B 细胞可能会被诱导分泌针对凋亡细胞 (ATAC) 的自身抗体，并破坏正常的清除机制，从而使自身免疫反应持续甚至扩大。或者，ATAC 可能继发于清除缺陷或凋亡细胞残余物过多。为了区分这些可能性，需要有关 ATAC 的遗传学、调控和功能的更详细信息。该提案旨在从凋亡细胞吞噬功能缺陷的小鼠中获得 ATAC。具体目的是：1)检测Mer、Axl、Tyro3三合子小鼠血清中ATAC的表达，并构建阳性小鼠杂交瘤； 2) 通过流式细胞术和共聚焦显微镜筛选杂交瘤的 ATAC 特异性，并调查它们与来自相同融合的其他自身反应性杂交瘤的关系； 3)确定ATAC特异性的遗传基础，并比较免疫球蛋白重链和轻链V基因使用、连接多样性和体细胞突变对凋亡细胞识别的贡献。未来的研究将探讨携带ATAC的B细胞对针对核和细胞表面抗原的自身免疫反应的贡献、可溶性ATAC对巨噬细胞清除凋亡细胞的影响以及携带ATAC的B细胞的调节。这些研究的结果将有助于更全面地评估凋亡细胞在诱导 B 细胞耐受或自身免疫中的作用。

项目成果

Heterogeneous nuclear ribonucleoprotein P2 is an autoantibody target in mice deficient for Mer, Axl, and Tyro3 receptor tyrosine kinases.

异质核核糖核蛋白 P2 是 Mer、Axl 和 Tyro3 受体酪氨酸激酶缺陷小鼠的自身抗体靶点。

• DOI: 10.4049/jimmunol.176.1.68
• 发表时间: 2006
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Radic,MarkoZ;Shah,Kinjal;Zhang,Wenguang;Lu,Qingxian;Lemke,Greg;Hilliard,GeorgeM
• 通讯作者: Hilliard,GeorgeM