HIGH MOLECULAR PROTEINS RESPONSIBLE FOR INTRACTABLE INFLAMMATORY DISEASES-IDENTIFICATION AND GENE EXPRESSION-

负责难治性炎症疾病的高分子蛋白-鉴定和基因表达-

• 批准号: 02454166
• 负责人: NOSE Masato
• 金额: $ 3.39万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454166/
• 关键词: Collagen disease; Autoimmune disease; Glomerulonephritis; Arteritis; Nephritogenic antibody genes; TNF; RFLP; Background genes; 骨炎惹起性抗体遺伝子; 腎炎惹起性単ク-ロン抗体; IgG3塩基配列; SCIDマウス; 腎炎惹起性単クロ-ン抗体

It is still unclear what functional molecules are critical for the development of intractable inflammatory diseases including collagen disease, although several parameters of autoimmunity have been pointed out in these diseases. This project was performed to identify high molecular proteins responsible for the development of collagen disease and to analyze their gene expression by using a murine model, MRL/Mp-lpr/lpr (MRL/lpr) mice. This strain of mice spontaneously develops a lethal glomerulonephritis, arteritis and arthritis, associated with the expression of the immunological disorder-inducing gene, lpr, which is recently clarified to be Fas antigen deletion mutant. These mice develop all of these diseases in the same individual, but MRL genetic background is required for them in addition to the lpr gene.We clarified that the background genes for these diseases are able to be genetically segregated each other by using the MRL hybrid mice with non-autoimmune-prone mice. Considering t … More he facts from these MRL hybrid mice, we identified the responsible proteins for collagen disease in MRL/lpr mice; IgG3 subclass for glomerulonephritis and TNF for granulomatous arteritis.In further analyses of IgG3, we succeeded in obtaining nephritogenic IgG3 producing clones against normal or SCID mice, and moreover these clone generated regular variations of lupus nephritis in histopathological manifestations. Immunoglobulin gene analysis of these clones clarified that each nephritogenic IgG3 is derived from a different B cell precursor and used a different VH germline gene. Moreover, one of these IgG3 was not associated with somatic mutation in the VH region, indicating that somatic mutation in the VH region is not required for the development of nephritogenic antibody. Furthermore, this germline VH gene was identical to that found in a non-autoimmune-prone mice. Thus, there may be no autoimmune-prone specific al- lelism in the germline VH gene relevant to the nephritogenic antibody.Regarding TNF on the development of arteritis, there is no RFLP of TNF-exon 4 among MRL and non- autoimmune-prone mice. However, it was clear that the individuals with arteritis among the MRL hybrid mice manifest a positive correlation between TNF and IL-1 beta mRNA level, but not between other macrophage-related cytokine mRNA such as LIF, M-CSF and Eta-1. This fact indicates that particular potential of macrophages are required for the development of arteritis in MRL/lpr mice, restricted with their background genes.Further studies of the sequences in the variable regions of IgG3 responsible for the nephritogenicity, including the association of the constant regions, and the induction mechanisms of nephritogenic antibody- producing B cell clones are required. An arteritis-prone strain of mice established from the MRL hybrid mice will be useful for further studies of the genetic basis of arteritis and of the responsible genes. Less

目前还不清楚哪些功能分子对包括胶原病在内的难治性炎症性疾病的发展至关重要，尽管已经指出了这些疾病中的几个自身免疫参数。本项目旨在通过使用小鼠模型MRL/Mp-lpr/lpr（MRL/lpr）小鼠来鉴定负责胶原病发展的高分子蛋白质并分析其基因表达。该品系小鼠自发地发展致死性肾小球肾炎、动脉炎和关节炎，与免疫紊乱诱导基因lpr的表达相关，lpr最近被澄清为Fas抗原缺失突变体。这些小鼠在同一个体中发生所有这些疾病，但是除了lpr基因之外，它们还需要MRL遗传背景。我们阐明了这些疾病的背景基因能够通过使用MRL杂交小鼠与非自身免疫易感小鼠进行遗传分离。考虑到t ...更多信息 从MRL/lpr小鼠中鉴定出与胶原病有关的蛋白质，即肾小球肾炎的IgG 3亚类和肉芽肿性动脉炎的TNF亚类，进一步分析IgG 3，成功地获得了抗正常或SCID小鼠的致肾炎IgG 3克隆，而且这些克隆在组织病理学上产生了狼疮性肾炎的规律性变化。这些克隆的免疫球蛋白基因分析阐明，每个致肾炎IgG 3来源于不同的B细胞前体，并使用不同的VH种系基因。此外，这些IgG 3之一与VH区的体细胞突变无关，表明VH区的体细胞突变对于致肾炎抗体的产生不是必需的。此外，该生殖系VH基因与在非自身免疫易感小鼠中发现的基因相同。关于TNF在动脉炎发生中的作用，在MRL和非自身免疫易感小鼠中不存在TNF-外显子4的RFLP。然而，很明显，在MRL杂交小鼠中患有动脉炎的个体表现出TNF和IL-1 β mRNA水平之间的正相关性，但其他巨噬细胞相关细胞因子mRNA之间没有相关性，如LIF、M-CSF和Eta-1。这一事实表明，MRL/lpr小鼠动脉炎的发生需要巨噬细胞的特殊潜能，受其背景基因的限制，需要进一步研究负责致肾炎性的IgG 3可变区序列，包括恒定区的关联，以及致肾炎性抗体产生B细胞克隆的诱导机制。从MRL杂交小鼠建立的动脉炎易感小鼠品系将有助于进一步研究动脉炎的遗传基础和相关基因。少

项目成果

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Takahashi,S.,etal.：“MRL/lpr 小鼠中 IgG3 的产生是狼疮性肾炎发生的原因”J.Immunol.147。

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Itoh, J.、Kinjoh, K.、Ohyama, A.、Nose, M. 和 Kyogoku, M.：“应用双色免疫荧光染色来演示类风湿性滑膜炎中的 T 细胞和 HLA-DR 携带细胞