Fibrinogen-endothelial cell interaction in vitro: a pathway mediated by an Arg-Gly-Asp recognition specificity.
It has been previously shown that fibrinogen (FG) associates specifically with human umbilical vein and bovine aortic endothelial cells (EC) in culture and induces EC migration. In the present study, we have investigated whether the FG-EC interaction is mediated by an Arg-Gly-Asp (RGD) recognition specificity and whether EC membrane proteins related to platelet GPIIb-IIIa are involved. Highly purified radioiodinated human FG, containing no detectable fibronectin, interacted with cultured human and bovine EC in suspension in a time-dependent and specific manner. The binding was inhibited by EDTA. Two polyclonal antibodies to platelet GPIIb-IIIa, which immunoprecipitated a heterodimer molecule from EC, inhibited FG binding to EC. These same antibodies inhibited FG-induced EC migration in a dose-dependent manner as measured in a Boyden chamber. Preabsorption of the antibodies with purified platelet GPIIb-IIIa markedly reduced both inhibitory activities. A series of synthetic RGD-containing peptides inhibited FG binding to EC and FG-induced EC migration. Gly-Arg-Gly-Asp (GRGD) was the most active peptide tested in inhibiting FG binding and EC migration (ID50 of 30 microM), and conservative substitutions in the RGD sequence markedly reduced inhibitory activity (ID50 greater than 1,000 microM). These results indicate that FG binding and EC migration are events mediated by an RGD recognition specificity and that EC surface proteins immunologically related to the GPIIb-IIIa complex on platelets are involved in this recognition.
Languino
L R
LR
Institute of Pharmacological Research Mario Negri, Milan, Italy.
Colella
S
S
Zanetti
A
A
Andrieux
A
A
Ryckewaert
J J
JJ
Charon
M H
MH
Marchisio
P C
PC
Plow
E F
EF
Ginsberg
M H
MH
Marguerie
G
G
D000954:Antigens, Surface / Q000502:physiology*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules; D002465:Cell Movement / Q000187:drug effects; D004730:Endothelium, Vascular / Q000502:physiology*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D008970:Molecular Weight; D009842:Oligopeptides / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013997:Time Factors
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D005340:Fibrinogen