1997 Fiscal Year Final Research Report Summary
Membrane inhibitors of complement in experimental animals
| Project/Area Number |
08044311
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
Immunology
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| Research Institution | Nagoya City University |
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Principal Investigator |
OKADA Hidechika Nagoya City Univ.Sch.Med., Dep.Mol.Biol., Prof., 医学部, 教授 (30160683)
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| Co-Investigator(Kenkyū-buntansha) |
LATZKOVITS Laszlo A.St.-Gyorgyi, Med.Univ., Neuroscience, Assoc.Prof., 助教授
MORGAN B.Paul Univ.Wales College Med., Dep.Med.Biochem., Prof., 医学部, 教授
LAJOS Baranyi Choju Med.Inst., Res.Fellow, 長寿医学研究所, 主任研究員 (10244542)
AKATSU Hiroyasu Choju Med.Inst., Res.Fellow, 長寿医学研究所, 研究員
OKADA Noriko Nagoya City Univ.Sch.Med., Dep.Mol.Biol., Asist.Prof., 医学部, 助手 (20160682)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Complement / Animals / DAF / MCP / CD59 / 512Ag / Crry / C5a receptor |
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| Research Abstract |
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.
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