MANIPULATING MHC GENE EXPRESSION IN ES CELL CHIMERAS

操纵 ES 细胞嵌合体中的 MHC 基因表达

基本信息

批准号：
3327189
负责人：
ELIZABETH K BIKOFF
金额：
$ 13.48万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1989
资助国家：
美国
起止时间：
1989-08-01 至 1992-11-30
项目状态：
已结题

项目摘要

The broad objective of this research is to analyse developmentally regulated expression of Class I products of the Major Histocompatability Complex in the mouse. In particular, the possibility that control of MHG gene expression may play a key role in maternal tolerance of the fetal allograft will be examined. We have designed a strategy that will allow us to study the consequences of ectopic expression of H-2D delta transplantation antigens in the developing embryo. A construct carrying the human Beta-actin gene promoter fused to the coding regions of the H-2D delta gene has been introduced into pluripotent embryonic stem (ES) cells. Although H-2D delta protein was not expressed at the cell surface in undifferentiated ES cells, significant amounts of H-2D delta membrane glycoproteins are detected on ES cells induced to differentiate in vitro. A major aim of the proposed investigation is to determine whether these transfected ES cell lines can contribute to all the somatic tissues when incorporated into normal embryos. Should our efforts to obtain live born chimeras be successful, the next goal will be to determine whether the introduced H-2D Delta gene can be transmitted in the germ line. Should transgenic chimeras fail to develop to term, we will describe the developmental failure and determine whether the abnormal phenotype (s) is attributable to defects in specific cell lineages. The spatial and temporal expression of Class I antigens in the developing embryos will be examined using immunohistochemical and in situ hybridization techniques. We will investigate the possibility that ectopic MHC gene expression in the extra- embryonic lineages may trigger a maternal immune response directed against the fetal allograft. A particularly important point will be to test whether maternal lymphocytes infiltrate and disrupt the abnormal embryos. These experiments will hopefully lead to a clearer understanding regulation of MHC gene expression during early mouse development and processes involved in maternal-fetal interactions.

这项研究的广泛目的是分析发展主要组织兼容性I类产品的调节表达在鼠标中复杂。特别是控制MHG的可能性基因表达可能在胎儿的孕产妇耐受性中起关键作用同种异体移植将检查。我们设计了一种策略，可以使我们研究H-2D三角洲异位表达的后果发育中的胚胎中的移植抗原。一个携带人β-肌动蛋白基因启动子融合到H-2D的编码区域 Delta基因已被引入多能胚胎（ES）细胞中。尽管H-2D三角洲蛋白在细胞表面未表达未分化的ES细胞，大量的H-2D三角洲膜在诱导分化体外的ES细胞上检测到糖蛋白。一个拟议调查的主要目的是确定这些是否是否当转染的ES细胞系可能有助于所有体细胞组织掺入正常胚胎中。我们应该努力获得诞生嵌合体成功，下一个目标是确定是否是否引入的H-2D三角洲基因可以在种系中传播。应该转基因嵌合体无法发展到学期，我们将描述发育失败并确定异常表型是否为归因于特定细胞谱系中的缺陷。空间和时间将检查发育胚胎中I类抗原的表达使用免疫组织化学和原位杂交技术。我们将研究异位MHC基因表达在外部的可能性胚胎谱系可能会触发针对的母体免疫反应胎儿同种异体移植。一个特别重要的一点是测试是否母体淋巴细胞浸润并破坏异常的胚胎。这些实验将有望导致更清楚的了解在早期小鼠发育和涉及过程中的MHC基因表达在母亲互动中。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Defective assembly of class I major histocompatibility complex molecules in an embryonic cell line.

胚胎细胞系中 I 类主要组织相容性复合体分子的组装有缺陷。

DOI：
10.1002/eji.1830210905
发表时间：
1991
期刊：
European journal of immunology
影响因子：
5.4
作者：
Bikoff,EK;Otten,GR;Robertson,EJ
通讯作者：
Robertson,EJ

Developmental failure of chimeric embryos expressing high levels of H-2Dd transplantation antigens.

表达高水平 H-2Dd 移植抗原的嵌合胚胎发育失败。

DOI：
10.1073/pnas.89.13.5927
发表时间：
1992
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Jaffe,L;Robertson,EJ;Bikoff,EK
通讯作者：
Bikoff,EK

Peptide and beta 2-microglobulin regulation of cell surface MHC class I conformation and expression.

DOI：
10.4049/jimmunol.148.12.3723
发表时间：
1992-06
期刊：
Journal of immunology
影响因子：
4.4
作者：
Gillis R. Otten;E. Bikoff;R. Ribaudo;Steven Kozlowski;David H. Margulies;Ronald N. Germain
通讯作者：
Gillis R. Otten;E. Bikoff;R. Ribaudo;Steven Kozlowski;David H. Margulies;Ronald N. Germain

Distinct patterns of expression of MHC class I and beta 2-microglobulin transcripts at early stages of mouse development.