Research for a retrovirus in myasthenic thymus and study for the pathogenesis

肌无力胸腺逆转录病毒的研究及发病机制研究

基本信息

批准号：
02454248
负责人：
ONO Akira
金额：
$ 2.24万
依托单位：
The Kitasato Institute
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-02454248/
关键词：
myasthenia gravis thymus thymoma retrovirus 遺伝子クロ-ニングレトロウィルス

项目摘要

By electron microscopy, we have demonstrated in retrovirus particles in cultured thymus cells (thymomas and thymus hyperplasias) of patients with myasthenia gravis. In most cases, these particles were detected in thymus epithelial cells when cultured with mitomycin C-treated human B-cells. For further characterization of these particles, virus-producer cell lines were established by co-culturing human T-lymphoblastoid cells with B-cell-stimulated human thymus cells. Mn^<++>-dependent RNA- directed DNA polymerase activity was detected at a density of 1.15-1.17 g/cm^3 on sucrose density gradients in the culture fluid of these co-cultured T-cells. We molecularly cloned the cDNA prepared by the detergent-activated endogenous polymerase reaction in the culture fluid of KK-ll cells, one of the virus-producer T-cell lines. The ^<32>p-labelled cloned cDNA hybridized to DNAs from freshly prepared hyman thymoma cells, in addition to KK-ll cell DNA. We sequenced about 400-1200 bp of seven cDNA clones, and the sequence of each cDNA was compared with that of all genes with known sequences present in EMBL data base. No striking similarity was observed when compared to other sequences. The nucleotide sequence of seven cloned cDNAs constantly revealed high homology (50-70%) with that of retroviruses such as HIV-l and equine infectious anemia virus (EIAV). These results indicate that cDNAs obtained here are new viral genes, possibly retroviral genes, derived from human thymus cells.

通过电子显微镜，我们已经在疗程的疗程性胸腺细胞（胸腺瘤和胸腺增生性增生性疗法）患者中证明了疗程疗程疗程的疗程。在大多数情况下，当用丝裂霉素C处理的人B细胞培养时，在胸腺上皮细胞中检测到这些颗粒。为了进一步表征这些颗粒，通过与B细胞刺激的人胸腺细胞共同培养人T-蛋白细胞细胞来建立病毒产生细胞系。 Mn^<++> - 依赖性RNA定向的DNA聚合酶活性在这些共培养T细胞的培养液中以1.15-1.17 g/cm^3的密度在1.15-1.17 g/cm^3中检测到。我们将通过洗涤剂激活的内源性聚合酶反应制成的cDNA在KK-LLL细胞的培养液中，这是病毒构成器T细胞系之一。 ^<32> p标记的克隆cDNA除了新鲜制备的海曼胸腺瘤细胞，除了KK-LLL细胞DNA外，还与DNA杂交。我们对七个cDNA克隆进行了约400-1200 bp的测序，并将每个cDNA的序列与所有具有EMBL数据库中存在已知序列的基因的序列进行了比较。与其他序列相比，没有观察到惊人的相似性。七个克隆cDNA的核苷酸序列与HIV-L和马匹感染性贫血病毒（EIAV）等逆转录病毒的核苷酸序列不断揭示出高的同源性（50-70％）。这些结果表明，这里获得的cDNA是源自人胸腺细胞的新病毒基因，可能是逆转录病毒基因。