Expression of functional decay-accelerating factor (CD55) in transgenic mice protects against human complement-mediated attack.
Transgenic mice expressing human CD55 were generated by microinjection of a CD55-minigene under the control of the mouse H2K(b) (MHC class I) promoter. Offspring were tested for transgene integration by PCR analysis, and for CD55 expression on peripheral blood leukocytes (PBLs) by flow cytometry. Expression levels of 15 founders ranged from 30 to 80% of that on human neutrophils. Immunohistochemical analysis of kidney, heart, liver, and lung tissue demonstrated staining for CD55 on endothelial surfaces as well as general diffuse staining throughout the tissues. The capacity of the transgenically expressed CD55 to prevent human C3 deposition on the surface of mouse splenocytes was assessed by flow cytometry. Cells from hemizygous mice incubated with 10% fresh human serum as a source of natural antibody and complement bound approximately 65% less C3 than control littermates. No further protection was seen using cells from homozygous littermates, and the protective effect was abrogated by prior incubation with an OFFi-CD55 monoclonal antibody. Similarly, transgenic mice were afforded significant protection from human serum-mediated lysis, determined using an LDH release assay. Hearts perfused with human plasma showed no increase in survival time in a modified Langendorff perfusion system, however deposition of human C3c was greatly reduced in transgenic hearts.
van Denderen
B J
BJ
Immunology Research Centre, St. Vincent's Hospital, Fitzroy, Victoria, Australia.
Pearse
M J
MJ
Katerelos
M
M
Nottle
M B
MB
Du
Z T
ZT
Aminian
A
A
Adam
W R
WR
Shenoy-Scaria
A
A
Lublin
D M
DM
Shinkel
T A
TA
d'Apice
A J
AJ
D000818:Animals; D001483:Base Sequence; D018958:CD55 Antigens / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology*; D003167:Complement Activation / Q000502:physiology*; D015932:Complement C3c / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic / Q000502:physiology*; D018014:Gene Transfer Techniques; D006183:H-2 Antigens / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008845:Microinjections; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism; D010477:Perfusion; D011401:Promoter Regions, Genetic; D013154:Spleen / Q000378:metabolism; D019076:Transgenes; D014183:Transplantation, Heterologous
D018958:CD55 Antigens; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D015932:Complement C3c