Molecular biological study of viruses in type 1 disabetes mellitus

1 型糖尿病病毒的分子生物学研究

• 批准号: 04454562
• 负责人: HANAFUSA Toshiaki
• 金额: $ 4.42万
• 依托单位: Osaka 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454562/
• 关键词: type 1 diabetes mellitus; genetics; autoimmunity; virus; NOD mouse; insulitis; retrovirus; cloning; gagタンパク; 膵臓; RT-PCR; 分子生物学

Type 1(insulin-dependent) diabetes mellitus is thought to develop through the mechanisms involving genetics, autoimmunity and viruses. Following our previous reports on the presence of virus-like particles in the NOD mouse (an animal model for type 1 diabetes mellitus), we intended to clone the virus genome and study its expression. NOD mice of 4,6 and 12 weeks of age were examined. Control mice included NON, ICR and B10.G.D.mice. RT-PCR method was used to clone retrovirus cDNA from RNA of the NOD mouse pancreas. PCR cloning was then performed to obtain the retrovirus genome. As a result, we succeeded in cloning the NOD mouse-specific retrovirus (NODV), the sequence of which has not been reported yet. Detailed analysis of NODV revealed following ; NODV is an intrinsic virus incorporated in the NOD mouse genome, its mRNA is expressed only in the pancreas of the NOD mouse, the highest expression of the mRNA is found at 4 weeks of age, i.e.the time when insulitis starts, and LTR of NODV has transcriptional activity in NIT-1 cells derived from NOD mouse beta cells. These results demonstrated the structure and expression of the virus which is thought to be involved in the development of insulitis and diabetes in the NOD mouse. Further analysis of the function of the NODV would disclose the role of viruses in the pathogenesis of type 1 diabetes mellitus.

1型(胰岛素依赖型)糖尿病被认为是通过遗传、自身免疫和病毒等机制发展起来的。根据我们之前关于NOD小鼠(1型糖尿病的动物模型)中存在病毒样颗粒的报道，我们打算克隆病毒基因组并研究其表达。取4周龄、6周龄和12周龄NOD小鼠。对照组为NON、ICR和B10.G.D.小鼠。用RT-PCR方法从NOD小鼠胰腺组织的RNA中克隆逆转录病毒基因。然后进行聚合酶链式反应克隆以获得逆转录病毒基因组。因此，我们成功地克隆了NOD小鼠特异性逆转录病毒(NODV)，其序列尚未见报道。对NODV的详细分析表明：NODV是一种整合在NOD小鼠基因组中的内源性病毒，其mRNA只在NOD小鼠的胰腺中表达，在4周龄时表达最高，即在NOD小鼠β细胞来源的NIT-1细胞中具有转录活性。这些结果证实了病毒的结构和表达，该病毒被认为与NOD小鼠的胰岛素炎和糖尿病的发展有关。进一步分析NODV的功能将揭示病毒在1型糖尿病发病机制中的作用。

项目成果

N.Itoh et al: "Mononuclear cell infiltration and its relation to the expression of MHC antigens and adhesion molecules in pancreas biopsy specimens from newly diagnosed insulin-dependent diabetes mellitus patients." J Clin Invest. 92. 2313-2322 (1993)

N.Itoh 等人：“新诊断的胰岛素依赖型糖尿病患者的胰腺活检标本中单核细胞浸润及其与 MHC 抗原和粘附分子表达的关系。”

T.Hanafusa et al.: "Diabetes in Japan" Elsevier Science Publishers B.V., (1994)

T.Hanafusa 等人：“日本的糖尿病”Elsevier Science Publishers B.V.，（1994 年）

K.Yamagata: "Overexpression of HLA class I antigen reduces insulin secretion in pancreatic b cells(RINm5F):evidence for non-immune mechanism." Res Commun Chem Pathol Pharmacol. 81. 299-308 (1993)

K.Yamagata：“HLA I 类抗原的过度表达会减少胰腺 b 细胞 (RINm5F) 中的胰岛素分泌：非免疫机制的证据。”

K.Yamamoto et al.: "Endothelial and microvascular abnormalities in the islet of non-obese diabetic (NOD) mice: transmission and scanning electron microscopic studies." Biomed. Res.13. 259-267 (1992)

K.Yamamoto 等人：“非肥胖糖尿病 (NOD) 小鼠胰岛的内皮和微血管异常：透射和扫描电子显微镜研究。”

N.Itoh et al.: "Transthyretin (prealbumin) in the pancreas and sera of newly diagnosed type l (insulin-dependent) diabetic patients." J.Clin.Endocrinol.Metab.74. 1372-1377 (1992)

N.Itoh 等人：“新诊断的 l 型（胰岛素依赖型）糖尿病患者的胰腺和血清中的甲状腺素运载蛋白（前白蛋白）。”