Mechanisn of lymphocyte differentiation, molecular mechanisms of gene rearrangement and clonal deletion by antigenes

淋巴细胞分化机制、反基因基因重排和克隆缺失的分子机制

基本信息

批准号：
04102007
负责人：
HONJO Tasuku
金额：
$ 117.76万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Specially Promoted Research
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

项目摘要

This year we have extended our previou studies using signal sequence trap method which allowed us to isolate a number of novel growth factors and their receptors. We are currently producing protein products encoded by isolated genes. These products will be added to the in vitro coculture system which will be described bellow. We have developed the new coculture system which allows to differentiate ES cells into all lineages of hematopoietic cells including lymphocytes. ES cells normally protected from differentiation by LIF was induced to differentiate by coculturing with a stroma cell line OP9 which cannot produce M-CSF because M-CSF inhibits differentiation into all the other hematopoietic lineages. Under this condition ES cells differentiate into red blood cells, granulocytes, megakaryocytes and lymphocytes. We have shown that lymphocytes actually express surface IgM.We have also shown several genes were induced upon programd cell death. One of such genes named MA3 was isolated and the sequence data indicate that MA3 is a novel gene with novel function. We have also isolated several known genes such as genes for heat shock protein. Another novel gene isolated in our laboratory about five years ago, colled RBP-Jk was originally assumed to be involved in immunoglobulin gene recombination because of the presence of the integrase-related motif. Knock out experiments and transgenic fry experiments indicate that the RBP-Jk gene is involved in signal transduction from the Notch receptor which regulates peripheral nervous development in Drosophila and probably lymphocyte proliferation in mammals. We have shown that RBP-Jk is involved in lateral inhibition of Drosophia and interacts with EBNA2 encoded by EB virus.

今年，我们使用信号序列捕获方法扩展了之前的研究，该方法使我们能够分离出许多新型生长因子及其受体。我们目前正在生产由分离基因编码的蛋白质产品。这些产品将被添加到体外共培养系统中，如下所述。我们开发了新的共培养系统，可以将 ES 细胞分化为包括淋巴细胞在内的所有造血细胞谱系。通常受 LIF 保护免于分化的 ES 细胞通过与基质细胞系 OP9 共培养而诱导分化，该基质细胞系 OP9 不能产生 M-CSF，因为 M-CSF 抑制分化为所有其他造血谱系。在此条件下，ES细胞分化为红细胞、粒细胞、巨核细胞和淋巴细胞。我们已经证明淋巴细胞实际上表达表面 IgM。我们还表明一些基因在程序性细胞死亡时被诱导。其中一个基因MA3被分离出来，序列数据表明MA3是一个具有新功能的新基因。我们还分离出了几个已知的基因，例如热休克蛋白基因。大约五年前我们实验室分离出的另一个新基因，colled RBP-Jk，由于存在整合酶相关基序，最初被认为参与免疫球蛋白基因重组。敲除实验和转基因鱼苗实验表明，RBP-Jk 基因参与 Notch 受体的信号转导，该受体调节果蝇的周围神经发育，也可能调节哺乳动物的淋巴细胞增殖。我们已经证明，RBP-Jk 参与果蝇的侧向抑制，并与 EB 病毒编码的 EBNA2 相互作用。