AUTOANTIGENS AND THE NUCLEAR BODY

真核细胞的成分被分成物理的和 被称为细胞器的功能隔间。一个特定的域或 核内的细胞器被指定为核体。这个 术语“核体”是指5-10个外接结构，0.2- 直径0.3微摩尔，分布在整个原子核 静息细胞。这项研究的长期目标是描述 并研究这种结构在人类疾病中的作用。 三条独立的调查路线表明，毒品调查科有权 进一步研究。L)NB(SP100)的一个组件是 40%的自身免疫性疾病患者的自身抗体 胆汁性肝硬变。相对较高的出现频率和特异性 PBC患者中针对该细胞器的抗体提示 提示NB可能参与了本病的发病机制。2)在 急性早幼粒细胞白血病(APL)患者发生易位 编码维甲酸受体α(RARpha)的基因和 编码核体第二组分的基因(PML)。PML- RARpha融合蛋白破坏NB并与 白血病细胞生长失调。3)在疱疹病程中 单纯疱疹病毒L(HSV-1)感染是一种与之相关的病毒蛋白(ICP0)， 并似乎扰乱了NB。病毒蛋白激活基因 转录，并参与潜伏病毒的重新激活。它是 预期拟议的研究将提供对 PBC、APL和HSV感染的病理生理学研究 在初步研究中，来自PBC患者的血清被用来识别 编码一种可能是第二自身抗原的新蛋白SP140的cDNA 在核弹体中。SP140的氨基酸序列与 SP100。此外，SP140还包含一个核酸结合基序，即 存在于另外两种已知与NB相关的蛋白质中 (PML和ICP0)。 该项目的具体目标包括：1)调查 SP140与核体的关系。多克隆抗血清 将针对SP140产生并用于确定位置 这种蛋白质在细胞中的含量。带有表位标签的编码SP140的cDNA 将被导入到真核细胞中，并且潜在的共刺激基因 将确定具有NB组分的SP140的本地化。2) 描述SP140与先前确定的 NB的组件。SP140将通过体外翻译和 SP140与NB的重组成分的相互作用将是 调查过了。 3)确定NB的其他组件。PBC患者的血清 将被用来识别编码NB中新自身抗原的cDNA。4) 确定针对自身抗体的临床意义 PBC患者脑白质成分的研究慢性支气管炎患者血清 将对有活检记录的PBC进行筛查，以检测针对该病毒的抗体 注意：

The components of eukaryotic cells are segregated into physical and functional compartments known as organelles. One specific domain or organelle within the nucleus has been designated the nuclear body. The term "nuclear bodies" (NBs) refers to 5-10 circumscribed structures, 0.2- 0.3 micromoles in diameter, that are distributed throughout the nucleus of resting cells. The long term objective of this study is to characterize the NB and investigate the role of this structure in human diseases. Three independent lines of investigation suggest that the NB warrants further study. l) A component of the NB (SP100) is a target of autoantibodies in 40% of patients with the autoimmune disease primary biliary cirrhosis. The relatively high frequency and specificity of antibodies directed against this organelle in patient with PBC suggest that the NB may be involved in the pathogenesis of this disease. 2) In patients with acute promyelocytic leukemia (APL), a translocation occurs between the gene encoding the retinoic acid receptor alpha (RARalpha) and a gene encoding a second component of the nuclear body (PML). The PML- RARalpha fusion protein disrupts the NB and is associated with dysregulated growth of leukemic cells. 3) During the course of herpes simplex virus l (HSV-1) infection, a viral protein (ICP0) associates with, and appears to disrupt, the NB. The viral protein activates gene transcription and is involved in reactivation of latent virus. It is expected that the proposed studies will provide insight into the pathophysiology of PBC, APL and HSV infection. In preliminary studies, serum from a patient with PBC was used to identify a cDNA encoding a novel protein, SP140, which may be a second autoantigen in the nuclear body. The amino acid sequence of SP140 is homologous to SP100. In addition, SP140 contains a nucleic acid-binding motif that is present in the two other proteins that are known to associate with the NB (PML and ICP0). The specific goals of this project include: 1) Investigate the relationship between SP140 and the nuclear body. Polyclonal antiserum directed against SP140 will be produced and used to determine the location of this protein in the cell. A cDNA encoding SP140 with an epitope tag will be transfected into eukaryotic cells and the potential co- localization of SP140 with components of the NB will be determined. 2) Characterize the interactions between SP140 and the previously identified components of the NB. SP140 will be prepared by in vitro translation and the interaction of SP140 with recombinant components of the NB will be investigated. 3) Identify additional components of the NB. Serum from patients with PBC will be used to identify cDNAs encoding new autoantigens in the NB. 4) Determine the clinical significance of autoantibodes directed against components of the NB in patients with PBC. Serum from patients with biopsy-documented PBC will be screened for antibodies directed against the NB.