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The development of xenograft

异种移植的发展

基本信息

批准号：
05557065
负责人：
SHIRAKURA Ryota
金额：
$ 12.86万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

We established several swine endothelial cell (SEC) lines, expressing human MCP,DAF,and an MCP/DAF hybrid by transfection of cDNA,and assessed the function of these transfectant molecules on C-mediated cell lysis as an in vitro hyperacute rejection model of swine to human discordant xenograft.1) DAF is quite effective to protect SEC from C-mediated cell lysis. Especially, over 80% suppression was observed in high expression clone. On the other hand, MCP is also effective in suppressing C activity, but the less efficient as the C3-step regulator than DAF.Hybrid showed approximate 60% of suppression of C-mediated cell lysis. It is more effective than MCP alone clearly, but indistinct from DAF alone because of the difference in the expression rate of transfectant molecules in each clone.2) CD59 is effective in protection of SEC when it is sufficiently expressed. However, DC59 is not so efficient in protecting SEC membrane from human C attack as DAF,because SEC is lysed at the C5b-8 step of the human complement cascade.TRANSGENICTo obtain the transgenic animals expressing human-MCP at high level, several studies concerning the promoter and enhancer have done. We obtained 11 independent lines of MCP transgenic mice lacking just the polyA signal and 5 lines without the entire 3'UT,as assessed by PCR and Southern blotting. While the expression of MCP in mice carrying MCP cDNA with 120bp of 3'UT was minimal, that in mice carrying MCP cDNA without total 3'UT was evident in many organs ; especially in muscle, heart, and pancreas.

我们建立了几个猪内皮细胞（SEC）系，表达人MCP，MCP，和MCP/MCP杂合体的cDNA转染，并评估这些转染分子对C介导的细胞溶解的功能，作为猪对人不协调异种移植物的体外超急性排斥模型。1）MCP是非常有效的保护SEC免受C介导的细胞溶解。特别是在高表达克隆中，观察到超过80%的抑制。另一方面，MCP在抑制C活性方面也是有效的，但是作为C3步骤调节剂的效率低于C3步骤调节剂。它明显比单独的MCP更有效，但由于每个克隆中转染分子表达率的差异，与单独的MCP的区别不明显。2）当CD 59充分表达时，它在SEC的保护中是有效的。然而，由于SEC在人补体级联反应的C5 b-8步骤被裂解，因此DC 59在保护SEC膜免受人C攻击方面不如人MCP有效。我们获得了11个单独的MCP转基因小鼠品系，仅缺乏polyA信号，5个品系没有整个3 'UT，如通过PCR和Southern印迹所评估的。尽管在携带有120 bp的3 'UT的MCP cDNA的小鼠中MCP的表达是最小的，但在携带MCP cDNA而没有总3' UT的小鼠中，MCP在许多器官中的表达是明显的，特别是在肌肉、心脏和胰腺中。