AUTOANTIGENS AND THE NUCLEAR BODY

真核细胞的组成部分被分离成物理的和 功能区称为细胞器。一个特定的域或 细胞核内的细胞器被称为核体。的 术语“核体”（NB）是指5-10个外切结构，0.2- 10个核体， 0.3微摩尔直径，分布在整个细胞核中， 休眠细胞本研究的长期目标是表征 NB并研究这种结构在人类疾病中的作用。 三个独立的调查线索表明，毒品和犯罪问题办公室 进一步研究l）NB（SP100）的组件是以下目标： 40%的自身免疫性疾病患者的自身抗体 胆汁性肝硬化相对较高的频率和特异性， PBC患者中针对该细胞器的抗体表明 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， 编码视黄酸受体α（RAR α）的基因与 编码核体（PML）的第二组分的基因。PML- RAR α融合蛋白破坏NB，并与 白血病细胞生长失调。3)在疱疹的过程中 单纯病毒I（HSV-1）感染，与之相关的病毒蛋白（ICP 0）， 似乎破坏了NB病毒蛋白激活基因 转录并参与潜伏病毒的再活化。是 预计拟议的研究将提供深入了解 PBC、APL和HSV感染的病理生理学。 在初步研究中，来自PBC患者的血清用于鉴定 编码新蛋白SP140的cDNA，其可以是第二自身抗原 在核体内。SP140的氨基酸序列与 SP100。此外，SP140含有核酸结合基序， 存在于已知与NB相关的另外两种蛋白质中 (PML ICP0）。 本项目的具体目标包括：1）调查 SP140与核小体的关系多克隆抗血清 将产生针对SP140的信号，并用于确定 这种蛋白质在细胞中的含量。一种编码带有表位标签的SP140的cDNA 将被转染到真核细胞中， 将确定SP 140与NB组件的定位。（二） 描述SP140与先前确定的 NB的组成部分。SP140将通过体外翻译制备， SP140与NB的重组组分的相互作用将是 研究了 3)确定NB的其他组件。PBC患者血清 将用于鉴定编码NB中新的自身抗原的cDNA。四、 确定针对以下疾病的自身抗体的临床意义 PBC患者的NB成分。患者血清 将对活检记录的PBC进行针对 NB.

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.