Generation of model mice for auto immune neuropathy by transfection with anti-carbohydrate chain antibody genes.

通过转染抗碳水化合物链抗体基因产生自身免疫神经病模型小鼠。

• 批准号: 09558099
• 负责人: TAI Tadashi
• 金额: $ 1.92万
• 依托单位: Tokyo Metropolitan Institute of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09558099/
• 关键词: monoclonal antibody; carbohydrate chain; ganglioside; transgenic mouse; auto-immune disease; neuropathy

The aim of the project is to establish model mice for auto-immune neuropathy by transfection with anti-carbohydrate antibody genes. A hybridoma producing a monoclonal antibody, which reacts specifically with b-pathway gangliosides such as GD3, GD2, GD1b, GT1b, and GQ1b, but not with any other gangliosides or neutral glycosphingolipids was selected among a series of hybridomas secreting monoclonal antibodies specific for carbohydrate chains, that were established in our laboratory. We generated, at first, transgenic mice having high antibody titer of IgM to the ganglioides. These mice appeared to be healthy and exhibited no significant pathological symptoms at 1-year-old. Next, we tried to induce neuropathy by immunizing these transgenic mice with a number of adjuvants including LPS and cytokines. However, they showed no significant symptoms. These results suggest that only high antibody titer to the gangliosides in sera is not enough for inducing neuronal symptoms. Further study will be needed for analyzing the mechanisms of the neuropathy. At present, we are producing transgenic mice having high antibody titer of IgG.

本课题的目的是通过抗糖抗体基因转染建立自身免疫性神经病小鼠模型。在我们实验室建立的一系列分泌糖链特异性单克隆抗体的杂交瘤中，选择了产生单克隆抗体的杂交瘤，该单克隆抗体与b途径神经节苷脂如GD3、GD2、GD1b、GT1b和GQ1b特异性反应，但不与任何其他神经节苷脂或中性鞘糖脂反应。首先，我们产生了对类神经节细胞具有高IgM抗体滴度的转基因小鼠。这些小鼠看起来很健康，在1岁时没有表现出明显的病理症状。接下来，我们尝试通过用包括LPS和细胞因子的许多佐剂免疫这些转基因小鼠来诱导神经病变。然而，他们没有表现出明显的症状。这些结果表明，仅血清中高滴度的神经节苷脂抗体不足以诱导神经元症状。神经病变的发生机制有待进一步研究。目前，我们正在生产具有高IgG抗体滴度的转基因小鼠。

项目成果

Ogura, K.: "Molecular cloning of a rat brain cDNA, with homology to a tyrosine kinase substrate, that induce galactosy ceramide expression and morphological changes in COS-7 cells"J. Neurochem.. 71. 1827-1836 (1999)

Ogura, K.：“大鼠脑 cDNA 的分子克隆，与酪氨酸激酶底物同源，可诱导 COS-7 细胞中半乳糖神经酰胺的表达和形态变化”J.

Liu, Y.: "A genetic model of substrate deprivation therapy for a glycosphingolipid storage disorder"J. Clin. Invest.. 103. 497-505 (1999)

Liu，Y.：“鞘糖脂贮积症底物剥夺疗法的遗传模型”J。

Sorice,M.: "Evidence for the existence of ganglioside enriched plasma membrane domains in human peripheral lymphocytes." J.Lipid.Res.38. 969-974 (1997)

Sorice,M.：“人外周淋巴细胞中存在富含神经节苷脂的质膜结构域的证据。”

Comas, T. C.: "Immunohistocemical staining for ganglioside GD1b as a diagnostic and prognositic marker for primary human brain"Neuro-Oncology. 1. 261-267 (1999)

Comas, T. C.：“神经节苷脂 GD1b 的免疫组织化学染色作为原发性人脑的诊断和预后标记”神经肿瘤学。

Sato,M.: "Accumulation of cholesterol and GM2 ganglioside in cells cultured in the presence of progesterone." Brain Dev.20. 50-52 (1998)

Sato,M.：“在存在黄体酮的情况下培养的细胞中胆固醇和 GM2 神经节苷脂的积累。”