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Pathogenetic roles of human endogenous retroviruses in autoimmune diseases

人内源性逆转录病毒在自身免疫性疾病中的致病作用

基本信息

批准号：
12670193
负责人：
IKEDA Hitoshi
金额：
$ 2.24万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670193/
关键词：
human endogenous retroviruses transgenic rat gene expression gene cloning autoimmune diseases

项目摘要

To investigate the pathogenetic roles of human endogenous retroviruses (HERVs) in autoimmune diseases, we produced transgenicrats carrying an HERV genome and analyzed the transgenic rat as a model. And we tried the cDNA gene cloning for novel HERVs from vascular endothelial cells as a target of autoimmune diseases and synovial tissues of rheumatoid arthritis. Results of this research project were described below.1. Two lines of transgenicrats carrying the full-length genome of HERV-R, which is a single copy type of HERV and expresses the env gene in several human tissues, were established. The transgene was specifically expressed at high levels in Harderian and submandibular salivary glands of the rats. Using a rabbit anti-HERV-R synthetic peptides antiserum, 85kDa of Env glycoprotein, contain about 23kDa of glycan, was detected in acinar cells of the Harderian glands. The transgene was consistently expressed in concepti from day 12 after gestation and was detected in fetuses at day 18 of gestation. By the skin transplantation experiment between the transgenic and non-transgenic rats, grafts of transgenic rats in non-transgenic rats were rejected by immunological reaction of host rats, suggesting that the 85kDa Env glycoprotein is recognized as an immunological target by non-transgenic rats. Although no disease was spontaneously developed in the transgenic rats, HERV-R-related immunological disorders may develop in the rats when the immunological tolerance against the 85kDa Env glycoprotein is broken.2. We screened cDNA libraries of human placentas, umbilical vein endothelial cells and synovial tissues of rheumatoid arthritis patients, using mixture of DNAs-related human retrovirus sequences as probes. Several clones with retrovirus sequences were detected, but we found no novel HERV sequence, yet. However, we evidenced that one HERV, which is not detected any transcription in normal tissues in previous reports, is expressed in several placentas without diseases.

为了研究人内源性逆转录病毒（HERV）在自身免疫性疾病中的致病作用，我们产生了携带 HERV 基因组的转基因大鼠，并将转基因大鼠作为模型进行分析。我们还尝试从血管内皮细胞中克隆新型 HERV 的 cDNA 基因，作为自身免疫性疾病和类风湿性关节炎滑膜组织的靶标。本课题的研究成果如下： 1．建立了两个携带 HERV-R 全长基因组的转基因鼠系，HERV-R 是 HERV 的单拷贝类型，并在多个人体组织中表达 env 基因。该转基因在大鼠的哈德氏唾液腺和颌下唾液腺中特异性高水平表达。使用兔抗HERV-R合成肽抗血清，在哈德氏腺腺泡细胞中检测到85kDa的Env糖蛋白，含有约23kDa的聚糖。该转基因从妊娠后第 12 天起在受孕者中持续表达，并在妊娠第 18 天的胎儿中检测到。通过转基因和非转基因大鼠之间的皮肤移植实验，转基因大鼠的移植物被宿主大鼠的免疫反应排斥，这表明85kDa Env糖蛋白被非转基因大鼠识别为免疫靶标。虽然转基因大鼠没有自发发生疾病，但当对85kDa Env糖蛋白的免疫耐受被破坏时，大鼠可能会出现HERV-R相关的免疫紊乱。 2．我们使用DNA相关的人逆转录病毒序列的混合物作为探针，筛选了人胎盘、脐静脉内皮细胞和类风湿关节炎患者滑膜组织的cDNA文库。检测到了几个带有逆转录病毒序列的克隆，但我们尚未发现新的 HERV 序列。然而，我们证明了一种HERV在一些没有疾病的胎盘中表达，这种HERV在之前的报道中在正常组织中没有检测到任何转录。