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The analysis of lymphoid cell differention by using cell lines transformed with a ts mutant of Abelson murine leukemia viurs.

使用 Abelson 鼠白血病病毒 ts 突变体转化的细胞系分析淋巴细胞分化。

基本信息

批准号：
61480154
负责人：
TAKEMORI Toshitaka
金额：
$ 4.1万
依托单位：
Chiba 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：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

We have succeeded in obtaining a ts mutant of Abelson murine leukemia viurs(A-MuLV) through a biologikal assay relying on changes in the differentiation state of B lymphomas at permissive and nonpermissive temperatures.The viral activity was further analysed by bone marrow colony assay, focus assay,and gene transfection assay using proviral DNA of ts virrus. The results confiem that the virus carries theproperty of a ts mutat virus.By using this ts mutant,designated as ts-49,we have established several immature B lymphomas to analyse the mechanism of immunoglobulin(Ig) expression during differentiation. We observed 2)the usage of a VH gene to join toa D@j segment was predetemined prior to the Ig rearrangement,2) u chains, synthesized at the stage of pre-B cells,were expressd on the cell surface as an associated from with unknown molecule(s).It is not known whether the phenomena are only peculiar to established lymphomas,however,the analysis provides new information about the programming of Ig gene expression,in contrast to the esteblished view that VH genes are rendomely selected and used for VDJ joining at the stage of Ig gene rearrangement.On the other hand,we have established a new system to transform lymphoid cells in fetal thymus by transforming viruses including a ts mutant.By using this new system,we observed that B precursors were stored at high frequency in fetal thymus.Fetal thymus carries activity. to store B precursors of fetal liver origin and to support the generation and survivals of pre-B lymphomas of thymic and fetal liver origin.This result leads new insight about the role of thymic stromas on B cell development.In addition,we have establishd immature T cell lines;one is "double negative",CD3+,and]positive cell and antogher is Thy-1+,AA4.1+, IL2R+.The latter may belong toimmature T lineage cells which are nou known in the literature.

我们根据B淋巴瘤在允许和非允许温度下分化状态的变化，成功地获得了Abelson小鼠白血病病毒的ts突变体（A-MuLV），并通过骨髓集落试验、病灶试验和ts病毒前病毒DNA的基因转染试验，进一步分析了病毒的活性。本研究利用ts-49突变株建立了几个未成熟B淋巴瘤，并分析了其分化过程中免疫球蛋白（IG）表达的机制。我们观察到2）在IG重排之前，VH基因与D@j片段的连接是预先确定的，2）在前B细胞阶段合成的u链在细胞表面表达，作为与未知分子相关的形式。目前还不知道这种现象是否仅在已建立的淋巴瘤中特有，但是，分析提供了关于IG基因表达程序的新信息，另一方面，我们建立了一个新的转化胎儿胸腺淋巴细胞的系统，通过转化含有ts突变体的病毒，观察到B前体在胎儿胸腺中以高频率储存，胎儿胸腺具有活性。胸腺基质对B细胞发育的作用有了新的认识，我们建立了未成熟T细胞系，并在此基础上建立了胸腺基质对B细胞发育的调控机制，为进一步研究胸腺基质对B细胞发育的调控机制提供了新的思路。一种为“双阴性”，CD 3+，]+细胞，另一种为Thy-1+，AA 4.1+，IL 2 R+细胞，后者可能属于未成熟的T细胞系。