| Co-Investigator(Kenkyū-buntansha) |
NAKAGAMI Tatsuyoshi Research and Development Center, Nippon Meat Products Inc., 中央研究所, 主任
SHIGEHISA Tamotsu Research and Development Center, Nippon Meat Products Inc., 中央研究所, 副主査
NAKATA Seizoh Osaka Univ Med School Assistant Professor, 医学部, 助手 (50116068)
SHIMAZAKI Yasuhisa Osaka Univ Med School Associate Professor, 医学部, 助教授 (60116043)
MATSUDA Hikaru Osaka Univ Med School Professor, 医学部, 教授 (00028614)
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| Research Abstract |
We established several swine endothelial cell (SEC) lines, expressing human MCP,DAF,and an MCP/DAF hybrid by transfection of cDNA,and assessed the function of these transfectant molecules on C-mediated cell lysis as an in vitro hyperacute rejection model of swine to human discordant xenograft.
1) DAF is quite effective to protect SEC from C-mediated cell lysis. Especially, over 80% suppression was observed in high expression clone. On the other hand, MCP is also effective in suppressing C activity, but the less efficient as the C3-step regulator than DAF.Hybrid showed approximate 60% of suppression of C-mediated cell lysis. It is more effective than MCP alone clearly, but indistinct from DAF alone because of the difference in the expression rate of transfectant molecules in each clone.
2) CD59 is effective in protection of SEC when it is sufficiently expressed. However, DC59 is not so efficient in protecting SEC membrane from human C attack as DAF,because SEC is lysed at the C5b-8 step of the human complement cascade.
TRANSGENIC
To obtain the transgenic animals expressing human-MCP at high level, several studies concerning the promoter and enhancer have done. We obtained 11 independent lines of MCP transgenic mice lacking just the polyA signal and 5 lines without the entire 3'UT,as assessed by PCR and Southern blotting. While the expression of MCP in mice carrying MCP cDNA with 120bp of 3'UT was minimal, that in mice carrying MCP cDNA without total 3'UT was evident in many organs ; especially in muscle, heart, and pancreas.
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