| Project/Area Number |
05454361
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General surgery
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| Research Institution | KYOTO PREFECTUAL UNIVERSITY OF MEDICINE |
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Principal Investigator |
OKA Takahiro Kyoto Prefectural University of Medicine, The second Department of Surgery, Professor, 医学部, 教授 (60079837)
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| Co-Investigator(Kenkyū-buntansha) |
ARAKAWA Kohei Kyoto Prefectural University of Medicine, The Second Department of Surgery, Assi, 医学部, 助手 (20167993)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1994: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | xenogeneic transplantation / complement inhibitory protein / CD59 / hyperacute rejection / immunosuppression / 超急性拒絶反応 / 補体制抑因子 / PCR法 / 免疫抑制 |
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| Research Abstract |
In Dicordant xenotransplantation, the most serious problem to be overcome is hyperacute rejection, in which complement play a major role on xenogeneic endothelial cells. Recently, we reported the introduction of CD59 to mouse fibroblast showed resistance against human complement attacking. The aim of this study is to investigate the role of N-glycosylation of CD59 antigen by site-directed mutagenesis. Second, we have evaluated the modification of the xenograft by introduction and expression of human CD59 gene in the canine kidney. Mutant CD59 expression vector, in which Ser-20 was changed to Ala, was constructed and introduced into a mouse A31 cell line. The functional analysis of the mutant CD59 expressed on mouse A31 cell line indicated that the mutant pSRalphaCD59/A31 was more registant to human complement attack than the pSRalphaCD59/A31 and pSR/A31. The molecular weight of wild-type and mutant CD59 revealed 20kd and 14kd, respectively. Using the hemagglutinating virus of Japan (HVJ) liposomes, which encapsulated both DNA and nonhistone chromosomal protein high-mobility group1 (HMG1), we evaluated thismixture by human CD59 gene transfer to the canine kidney. RT-PCR analysis indicated CD59 was well transcribed in canine kidney. Immunohistchemical analysis revealed that CD59 was detected in the canine renal glomerular cells. In coclusion, we beleive that such techniques for postnatal animals will provide a suitable xenograft for humans.
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