Aggregated IgG bind to glomerular epithelial cells to stimulate urokinase release through an endocytosis-independent process.
BACKGROUND
In membranous nephropathy, the development of glomerular lesions is related to the formation of immune complexes at subepithelial sites. These deposits are associated with modifications in the fibrinolytic activity of glomerular cells leading to the appearance of fibrin degradation products in the deposits and the urine. A previous study has shown that immune complexes interact with glomerular epithelial cells (GEC) through the neonatal Fc receptor (FcRn). We therefore determined whether this binding could be responsible for a modification in the fibrinolytic activity of GEC.


METHODS
Endocytosis of heat-aggregated immunoglobulins (AgIgG) in cultured human GEC was studied by immunofluorescence and confocal microscopy. The release of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor 1 (PAI) by GEC or whole glomeruli was assessed by ELISA, fibrin zymography and Northern blot.


RESULTS
Human GEC in culture bound AgIgG that possess characteristics similar to soluble immune complexes and internalized them by 10 min. This process was mediated by FcRn since chicken aggregated IgG (AgIgY), that do not bind FcRn, did not colocalize with AgIgG in GEC. AgIgG but not AgIgY induced a decrease of FcRn expression at the membrane and within the cells. The binding of AgIgG to GEC elicited a dose- and time-dependent increase in the release of uPA activity, as in the uPA protein and mRNA expression without modification in the release of PAI. This process was not abrogated by agents inhibiting endocytosis and/or transcytosis such as cytochalasin B, suggesting an endocytosis-independent uPA regulation.


CONCLUSIONS
GEC response to AgIgG overload comprises at least two sequential steps: (1) a FcRn-mediated endocytosis; (2) an endocytosis-independent fibrinolytic imbalance leading to plasmin generation which could favor in vivo AgIgG clearance and matrix remodeling.
Haymann
Jean-Philippe
JP
INSERM U 489, Université Paris 6 and Association Claude-Bernard, Hôpital Tenon, Paris, France. jean-philippe.haymann@tnn.ap-hop-paris.fr
Delarue
Françoise
F
Baud
Laurent
L
Sraer
Jean-Daniel
JD
D001666:Binding Sites, Antibody; D002460:Cell Line; D002478:Cells, Cultured; D004705:Endocytosis / Q000502:physiology*; D004847:Epithelial Cells / Q000201:enzymology / Q000276:immunology*; D015395:Histocompatibility Antigens Class I; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1; D011961:Receptors, Fc / Q000378:metabolism; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000378:metabolism*
D015395:Histocompatibility Antigens Class I; D007074:Immunoglobulin G; D017395:Plasminogen Activator Inhibitor 1; D011961:Receptors, Fc; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; C100688:Fc receptor, neonatal