Development of transgenic animals of complement regulatory molecules.

补体调节分子转基因动物的开发。

• 批准号: 07557337
• 负责人: OKADA Noriko
• 金额: $ 1.47万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557337/
• 关键词: xenotransplantation; complement; regulatory molecules; DAF; Crry; MCP; トランスジェニック動物; 異種移植; 種特異的補体制御膜因子; 補体活性化

Complement plays an essential role in the acute rejection of discordant xenotransplants. To overcome this problem, development of transgenic animals have been generated which have cDNA of the species specific membrane inhibitors of human. However, to detarmine its applicability to suppress the acute rejection of xenotransplants, experimental models of xenotransplantation between experimental animals.For this purpose, gene cloning of the species specific membrane inhibitors is an essential requirement. We have already cloned the cDNA of rat 512 antigen (512Ag) which restricts C3 convertase of rat complement. In this research project we have indentified guinea pig DAF (GP-DAF) with monoclonal antibody (mAb), and its cDNA has been successfully cloned demonstrating that GP-DAF is comprised of several isotypes due to the alternative splicing. Furthermore, we have successfully cloned cDNA of guinea pig MCP (GP-MCP), mouse DAF,mouse CD59 and rat DAF.Using these cDNA,we have prepared CHO cells transfected with cDNA of rat 512Ag, GP-DAF and GP-MCP,and these cells have been demonstrated to be registant to the serum complement homologous to the genes expressed on the transfectants.

补体在不一致的异种移植物的急性排斥中起着重要作用。为了克服这个问题，已经开发出具有人类物种特异性膜抑制剂的cDNA的转基因动物。然而，为了确定其抑制异种移植急性排斥反应的适用性，实验动物之间异种移植的实验模型。为此目的，物种特异性膜抑制剂的基因克隆是基本要求。我们已经克隆了限制大鼠补体C3转化酶的大鼠512抗原(512Ag)的cDNA。在本研究项目中，我们用单克隆抗体（mAb）鉴定了豚鼠DAF（GP-DAF），并且其cDNA已成功克隆，证明GP-DAF由于选择性剪接而由多种同种型组成。此外，我们还成功克隆了豚鼠MCP（GP-MCP）、小鼠DAF、小鼠CD59和大鼠DAF的cDNA。利用这些cDNA，我们制备了转染大鼠512Ag、GP-DAF和GP-MCP cDNA的CHO细胞，并且这些细胞已被证明对与转染子上表达的基因同源的血清补体具有注册性。

项目成果

Hayashi, S., Okada, H., Emi, N., Okada, N., Yokoyama, I.and Takagi, H.: "Protection of guinea pig cells from rat complement-mediated lysis by the transfection of rat complement regulatory factor 512 gene." Xenotransplanmtation. 3. 217-221 (1996)

Hayashi, S.、Okada, H.、Emi, N.、Okada, N.、Yokoyama, I. 和 Takagi, H.：“通过转染大鼠补体调节因子来保护豚鼠细胞免受大鼠补体介导的裂解”

Wu,X.: "IgM natural antibody against an asialo-oligosaccharide,gangliotetraose(Gg4)sensitize HIV-I infected cells for cytolysis by homologous complement." Int.Immunol.8. 153-158 (1996)

Wu, X.：“针对脱唾液酸寡糖、神经节四糖 (Gg4) 的 IgM 天然抗体使 HIV-1 感染细胞对同源补体的细胞溶解敏感。”

Nishikawa,K.: "Local inflammation caused by a monoclonal antibody which blocks the function of the rat membrane inhibitor of C3 convertase." J.Immunol.156. 1182-1188 (1996)

Nishikawa,K.：“由单克隆抗体引起的局部炎症，该抗体可阻断 C3 转化酶的大鼠膜抑制剂的功能。”

Fukuoka,Y.: "Molecular cloning of murine decay accelerating factor(DAF)by immunoscreening." Int.Immunol.8. 379-385 (1996)

Fukuoka,Y.：“通过免疫筛选对小鼠腐烂加速因子（DAF）进行分子克隆。”

Hosokawa, M.: "Molecular cloning of guinea pig membrane cofactor protein (MCP) : Preferential expression in testis." J. Immunol.157. 4946-4952 (1996)

Hosokawa, M.：“豚鼠膜辅因子蛋白 (MCP) 的分子克隆：在睾丸中优先表达。”