Protection of rat endothelial cells from primate complement-mediated lysis by expression of human CD59 and/or decay-accelerating factor.
The present study analyzed the ability of human decay-accelerating factor (DAF) and CD59 to protect rat endothelial cell (EC) clones from human and primate complement-mediated lysis. By flow cytometry and Scatchard analysis, we show that human DAF and/or CD59 cDNAs under the transcriptional control of elongation factor 1-alpha promoter were expressed at levels similar to or higher than that of a human EC line. Human DAF and CD59 were released from the surface of transfected rat cells by phosphatidylinositol phospholipase C, demonstrating that the two molecules were linked to the cell membrane by means of a glycolipid anchor. The functional activity of the two human C regulatory proteins expressed on rat EC lines was studied using an in vitro assay of C-dependent cytotoxic in which rat EC were incubated with human or nonhuman primate sera as sources of xenogeneic natural antibodies and C. We demonstrate that human and monkey xenogeneic natural antibodies bind to rat cells and induce lysis by a C-dependent mechanism involving mainly the C direct activation pathway. Our data indicate that human DAF and CD59, expressed either alone or in combination, abrogated all EC cytotoxicity, even in the presence of 50% human serum. This protective phenotype was correlated with decreased membrane attack complex (CD59 and/or DAF transfectants) and C3 deposition (DAF transfectants) on EC surface. Antibodies against the transfected molecules abolished the protection against C-mediated lysis.
Charreau
B
B
Institut National de la Santé et de la Recherche Médicale (INSERM U211, Centre Hospialier Universitaire, Hotel Dieu, Nantes, France.
Cassard
A
A
Tesson
L
L
Le Mauff
B
B
Navenot
J M
JM
Blanchard
D
D
Lublin
D
D
Soulillou
J P
JP
Anegon
I
I
D000818:Animals; D015703:Antigens, CD / Q000494:pharmacology*; D018958:CD55 Antigens; D018957:CD59 Antigens; D002470:Cell Survival; D002472:Cell Transformation, Viral; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000187:drug effects / Q000276:immunology; D004730:Endothelium, Vascular / Q000166:cytology*; D005453:Fluorescence; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D008252:Macaca fascicularis / Q000097:blood; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013539:Simian virus 40 / Q000502:physiology; D014162:Transfection; D014183:Transplantation, Heterologous / Q000502:physiology
D015703:Antigens, CD; D018958:CD55 Antigens; D018957:CD59 Antigens; D015938:Complement Membrane Attack Complex; D008562:Membrane Glycoproteins