Membrane inhibitors of complement in experimental animals

实验动物补体膜抑制剂

• 批准号: 08044311
• 负责人: OKADA Hidechika
• 金额: $ 5.06万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08044311/
• 关键词: Complement; Animals; DAF; MCP; CD59; 512Ag; Crry; C5a receptor; モルモット; ラット; C5_aレセプター; オカダ酸; 補体制御膜因子

Species specitic membrane inhibitors of complement such as DAF,MCP and HRF20 (CD59) were identified in experimental animals. We have previously purified guinea pig DAF and rat 512 Ag and cloned cDNA of the molecules. Since guinea pig DAF was found to have several isotypes due to alternative splicing of mRNA,cDNA of each isotypes were transfectedinto CHO cells and the function of those geneproducts on the transfected cells were analyzed. We found that the longer the S/T rich region, the stronger the inhibitory function was observed.We have also cloned cDNa of guinea pib MCP,mouse DAF,mouse MCP,mouse CD59, rat MCP,and rat DAF.Mouse DAFcDNA was transfectedinto CHO cells and monoclonal antibodies to mouse DAF were generated by immunizing the transfected CHO cells into hamuster and making hybridoma cells fusing the spleen cells with mouse myeloma cells. Tissue distribution of guinea pig DAF,mouse DAF and rat DAF was determined by immunostaining with monoclonal antibodies. RT-PCR methodand Western blotting methodwere also succfessfully used to determine the tissues expressing guinea pig DAF and MCP,mouse DAF,MCP and CD59 as well as rat DAF,MCP and CD59. Furthermore we have also cloned cDNA of anaphylatoxin receptors of mouse and rat.

在实验动物中鉴定出补体的物种特异性膜抑制剂，如 DAF、MCP 和 HRF20 (CD59)。我们之前已经纯化了豚鼠 DAF 和大鼠 512 Ag，并克隆了这些分子的 cDNA。由于豚鼠DAF由于mRNA的选择性剪接而具有多种同种型，因此将每种同种型的cDNA转染至CHO细胞中，并分析这些基因产物在转染细胞上的功能。我们发现S/T丰富区越长，抑制功能越强。我们还克隆了几内亚猪MCP、小鼠DAF、小鼠MCP、小鼠CD59、大鼠MCP和大鼠DAF的cDNA。将小鼠DAFcDNA转染到通过将转染的 CHO 细胞免疫至仓鼠并制备与脾细胞融合的杂交瘤细胞，产生 CHO 细胞和针对小鼠 DAF 的单克隆抗体与小鼠骨髓瘤细胞。通过单克隆抗体免疫染色测定豚鼠DAF、小鼠DAF和大鼠DAF的组织分布。 RT-PCR法和Western blotting法也成功检测了豚鼠DAF、MCP、小鼠DAF、MCP、CD59以及大鼠DAF、MCP、CD59表达的组织。此外，我们还克隆了小鼠和大鼠的过敏毒素受体的cDNA。

项目成果

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

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

Wang, G., et.al.: "Functional differences among multiple isoforms of guinea pig decay accelerating factor (DAF)." J.Immunol.(in press). (1998)

Wang, G. 等人：“豚鼠腐烂加速因子 (DAF) 多种亚型之间的功能差异。”

Baranyi, L., Campbell, W. and Okada, H.: "Antisense homology boxes in C5a receptor and C5a anaphylatoxin. A new method for identification of potentially active peptides." J. Immunol.157. 4591-4601 (1996)

Baranyi, L.、Campbell, W. 和 Okada, H.：“C5a 受体和 C5a 过敏毒素中的反义同源盒。一种鉴定潜在活性肽的新方法。”

Akatsu, H.et al.: "Unipue expression of HRF20 (CD59) in human nervous tissue." Microbiol.Immunol.41. 321-330 (1997)

Akatsu, H.et al.：“HRF20 (CD59) 在人类神经组织中的 Unipue 表达。”

Akatsu, H., Okada, N., Okada, H. et al.: "Unique expression of HRF20(CD59)in human nervous tissue." Microbiol. Immunol.41(in press). (1997)

Akatsu, H.、Okada, N.、Okada, H. 等人：“HRF20(CD59) 在人类神经组织中的独特表达。”