CD59 expressed by human endothelial cells functions as a protective molecule against complement-mediated lysis.
CD59 is a 18-20-kDa membrane glycoprotein that inhibits formation of the membrane attack complex of complement (C) on homologous cells. In the present study we analyzed the expression and function of CD59 on human endothelial cells. Immunohistochemical analysis of renal cortex demonstrated a predominant expression of CD59 on peritubular capillary endothelial cells and glomerular endothelial cells. Flow cytometry analysis showed that human umbilical vein endothelial cells (HUVEC) expressed CD59 and the fluorescence intensity was approximately four times that of peripheral blood lymphocytes. CD59 is detected on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a single 20-kDa molecule in 2% deoxycholate extracts of HUVEC. CD59 was released from the surface of HUVEC by phosphatidylinositol-specific phospholipase C, demonstrating that it is attached to the cell membrane by means of a glycolipid anchor. The functional activity of CD59 expressed on HUVEC was studied. Blocking of CD59 antigen with F(ab')2 fragments of polyclonal anti-CD59 enhanced markedly the susceptibility of HUVEC to C-mediated lysis. This effect was dependent on the amount of blocking antibodies added. Northern blot analysis revealed the presence of three species of mRNA expressed in HUVEC, which hybridized to a cDNA probe specific for CD59, with sizes of about 800, 1400 and 2000 bp. These findings suggest that CD59 may be important in protection of endothelial cells against C-mediated damage at local sites of inflammation, thereby maintaining the vascular integrity in vivo.
Department of Nephrology, University Hospital, Leiden, The Netherlands.
R A
RA
Brooimans
A A
AA
Van der Ark
M
M
Tomita
L A
LA
Van Es
M R
MR
Daha
D015703:Antigens, CD / Q000032:analysis* / Q000502:physiology; D018957:CD59 Antigens; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000502:physiology*; D003602:Cytotoxicity, Immunologic; D004730:Endothelium, Vascular / Q000276:immunology*; D006017:Glycolipids / Q000032:analysis; D017261:Glycosylphosphatidylinositols; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000502:physiology; D010716:Phosphatidylinositols / Q000032:analysis
D015703:Antigens, CD; D018957:CD59 Antigens; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D010716:Phosphatidylinositols; D003165:Complement System Proteins