The suppression of the delayed-type xenograft rejection

延迟型异种移植排斥的抑制

基本信息

批准号：
15390377
负责人：
MIYAGAWA Shuji
金额：
$ 9.41万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390377/
关键词：
CRP DAF Factor I Complement deposit glycoantigen α-Gal antigen pig islet α-1,3GT 補体抑制因子 MCP ブタislet

项目摘要

Complement regulatory proteinThe cell membrane-bound forms of hole factor I (fI-PI), the light chain of the serine protease (SP) domain (SP-PI), and the light chain plus the COOH-terminal 45 amino acid of the heavy chain (SP+45-PI) were constructed. CHO cell transfectants with SP-PI also showed a clear inhibition in cell lysis by human serum, whereas CHO cell transfectants with SP+45-PI showed no inhibition.Tandem DAFcDNAs of the delta-SCR1-DAF, the double-DAF, the triple-DAF, and the tetra-DAF were established. CHO cell lines and pig endothelial cell (PEC) line expressing these molecules were established. The clear tendency was found in which the higher the tandem number of DAF increase, the more it affects the complement-mediated lysis.DAF function for NK cellWe report on an investigation of the effect of DAF on NK cell-mediated cytolysis. Delta-SCR2-DAF and delta-SCR3-DAF failed to suppress both complement-mediated and NK-mediated PEC lyses. However, delta-SCR4-DAF showed a clear complement regulatory effect, but had no effect on NK cells. The data suggest DAF has the direct inhibitory function on NK cells via SCR2-4.The antigenicity of pig isletsAdult pig islets expressed negligible amounts of α-Gal epitope, and α1,3GT activity was also undetectable. However, α-Gal was clearly expressed on the neonatal porcine islet-like cell clusters (NPCCs) cell surface.Transgenic pig experimentsThe issue of whether islets from GnT-III transgenic pigs can prolong their survival in cynomolgus monkeys was examined. While adult pig islets isolated from GnT-III transgenic pigs survived longer than those from wild-type pigs in the cynomolgus monkey, NPCCs from GnT-III transgenic pigs indicated little prolongation.

补体调节蛋白细胞膜结合形式的空穴因子 I (fI-PI)、丝氨酸蛋白酶 (SP) 结构域 (SP-PI) 的轻链以及轻链加上重链 COOH 末端 45 个氨基酸构建了具有 SP-PI 的 CHO 细胞转染子链 (SP+45-PI)，对人血清的细胞裂解也显示出明显的抑制作用，而具有 SP+45-PI 的 CHO 细胞转染子则显示出抑制作用。建立了 delta-SCR1-DAF、双 DAF、三 DAF 和四 DAF 的 DAFcDNA，并建立了表达这些分子的猪内皮细胞（PEC）系。 DAF串联增加的数量越多，对补体介导的裂解的影响越大。DAF对NK细胞的功能我们报告了DAF对NK细胞介导的细胞裂解作用的研究。 Delta-SCR2-DAF 和 delta-SCR3-DAF 未能抑制补体介导的和 NK 介导的 PEC 裂解，但 delta-SCR4-DAF 显示出明显的补体调节作用，但对 NK 细胞没有影响。 DAF通过SCR2-4对NK细胞具有直接抑制作用。猪胰岛的抗原性成年猪胰岛表达可忽略不计的α-Gal表位，并且α1,3GT活性也检测不到，但α-Gal在新生猪胰岛样细胞簇(NPCC)细胞表面清晰表达。转基因猪实验GnT-III转基因猪的胰岛是否可以延长食蟹猴的存活时间。虽然从 GnT-III 转基因猪中分离出的成年猪胰岛在食蟹猴中的存活时间比从野生型猪中分离出来的更长，但NPCCs 的存活时间更长。来自 GnT-III 转基因猪的结果表明几乎没有延长。