Porcine MCP gene promoter directs high level expression of human DAF (CD55) in transgenic mice.
Porcine membrane cofactor protein (pMCP), a complement regulatory protein, is widely expressed in various tissues. Particularly, it is highly expressed on vascular endothelium. The objective of this study was to investigate whether the pMCP gene promoter can induce efficient expression of a human complement regulatory protein, decay-accelerating factor (DAF; CD55) in transgenic mice. Two fragments of the 5'-flanking region of pMCP gene (0.9 kb and 5.4 kb) connected with human DAF minigene (0.9/hDAF and 5.4/hDAF) were used to produce transgenic mice. The expression of hDAF in heart, liver, kidney, lung, pancreas, brain and testis of the transgenic mice was examined by immunohistochemical analysis. The vascular endothelia and the nerves in all organs examined were intensely stained. The staining pattern in these tissues was similar in all transgenic mice examined regardless of the length of the promoters. The surface expression levels of hDAF on peripheral red blood cells and splenocytes from a mouse carrying 5.4/hDAF hemizygously was twice the level of expression on corresponding human cells. The red blood cells and splenocytes from the transgenic mice exhibited resistance to lysis by human serum in a manner dependent upon expressed hDAF level. The hearts from the transgenic mice functioned for a significantly longer time than those from normal mice under perfusion with human serum in the Langendorff perfusion system. These results demonstrated that the pMCP gene promoter is a good candidate of the regulatory element in the transgene to produce transgenic animals for xenotransplantation.
Murakami
H
H
Research and Development Center, Nippon Meat Packers, Inc., Tsukuba, Ibaraki, Japan.
Takahagi
Y
Y
Yoshitatsu
M
M
Miyagawa
S
S
Fujimura
T
T
Toyomura
K
K
Shigehisa
T
T
Shirakura
R
R
Kinoshita
T
T
D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001921:Brain / Q000378:metabolism; D018958:CD55 Antigens / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D003165:Complement System Proteins / Q000494:pharmacology; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D006321:Heart / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010477:Perfusion; D011401:Promoter Regions, Genetic; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013552:Swine; D019076:Transgenes; D014781:Viscera / Q000098:blood supply / Q000294:innervation / Q000378:metabolism
D015703:Antigens, CD; C497857:CD46 protein, human; D018958:CD55 Antigens; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D003165:Complement System Proteins