Identification of human endogenous retroviral antigen and its association with diseases

人内源性逆转录病毒抗原的鉴定及其与疾病的关系

• 批准号: 03670184
• 负责人: MARUYAMA Naoki
• 金额: $ 1.34万
• 依托单位: Tokyo Metropolitan Institute of Gerontology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670184/
• 关键词: endogenous retrovirus; recombinant protein; natural antibody; collagen disease; 自然坑体; 内存性レトロウィルス; 組替え蛋白; SLE

To identify the presence of human endogenous retroviral gene product in human tissue, we prepared several recombinant proteins by using gene technology. One of env gene product derived from human endogenous retrovirus like genomic DNA was produced in E. coli. We purified this recombinant protein and immunized to rabbit produce antibody. By using antibody against recombinant env gene product we observed the presence of human endogenous retroviral antigen in the placenta. We detected natural antibody against this protein in the patient sera from systemic lupus erythematosus. Next we prepared recombinant protein using Baculovirus vector system. We modified the genomic DNA encoding gag region of human endogenous retrovirus like DNA by site mutagenesis to produce long-sized openreading frame. By using this modified DNA we obtained recombinat protein corresponding gag protein fo murine of feline leukemia virus. When developing Western hybridization of this recombinant protein with anti-murine leukemia viral p30 serum, the recombinat protein showed strong positivity. These results indicated the origin of human endogenous retrovirus like sequences are derived same origin with murine leukemia virus genome.Our results may help to understand the association between human endogenous retrovirus and human diseases in future.

为了鉴定人内源性逆转录病毒基因产物在人体组织中的存在，我们利用基因技术制备了几种重组蛋白。从人内源性逆转录病毒样基因组DNA中获得一种env基因产物。我们纯化了该重组蛋白，并免疫兔产生抗体。利用重组env基因产物的抗体，我们观察到胎盘中存在人内源性逆转录病毒抗原。我们在系统性红斑狼疮患者血清中检测到该蛋白的天然抗体。接下来，我们利用杆状病毒载体系统制备重组蛋白。我们通过点突变的方法对编码人内源性逆转录病毒类DNA的基因组DNA进行了修饰，得到了大小较长的开放阅读框。利用修饰后的DNA，我们获得了猫白血病病毒小鼠的重组蛋白对应的Gag蛋白。将该重组蛋白与小鼠白血病病毒P30抗血清进行Western杂交，结果显示重组蛋白呈强阳性。这些结果表明人类内源性逆转录病毒样序列的起源与小鼠白血病病毒基因同源，我们的结果可能有助于进一步了解人类内源性逆转录病毒与人类疾病之间的关系。

项目成果

SHIGEMOTO et al: "Expression and stracture of serum gp 70 as an acute phase protein in NZB micl" Molecular Immunology. 29. 573-582 (1992)

SHIGEMOTO 等人：“NZB micl 中血清 gp 70 作为急性期蛋白的表达和结构”分子免疫学。

Kitamura,M., Shirasawa,T., Mitarai,T., Muramatsu,T., and Maruyama,N: "A retinoic acid responsive gene, MK, is preferentially expressed in the proximal tubules of the kidney and human tumor cell lines" American J. Pathology.

Kitamura,M.、Shirasawa,T.、Mitarai,T.、Muramatsu,T. 和 Maruyama,N：“视黄酸反应基因 MK 优先在肾和人类肿瘤细胞系的近端小管中表达”

Shigemoto et al.: "Expression and structure of serum gp70 as an acute phase protein in NZB mice" Molecular lmmunology. 29. 573-582 (1992)

Shigemoto 等人：“NZB 小鼠中血清 gp70 作为急性期蛋白的表达和结构”分子免疫学。

Itoh,Y., Takeuchi,S., Shigemoto,K., Kubo,S., Handa,S., Ishikawa,N. and Maruyama,N: "The strain-dependent constitutive expression of murine serum amyloid-P component is regulated at the transcriptional level" Biochimica et Biophysica Acata. 1131. 261-269 (

伊藤，Y.，竹内，S.，茂本，K.，久保，S.，半田，S.，石川，N.

Itoh,Y et al.: "The strain-dependent constitutive expression of murine serum amyloid-P component is regulated at the transeriptional level." Biochimica et Biophysica Acata. 1131. 261-269 (1992)

Itoh,Y 等人：“鼠血清淀粉样蛋白 P 成分的菌株依赖性组成型表达在转录水平上受到调节。”