Cooperation of C1q receptors and integrins in C1q-mediated endothelial cell adhesion and spreading.
The interaction of C1q with endothelial cells elicits a multiplicity of biologic responses. Although these responses are presumed to be mediated by the interaction of C1q with endothelial cell surface proteins, the identity of the participants is not known. In this study we examined the roles of two C1q binding proteins, cC1q-R/calreticulin and gC1q-R/p33, in C1q-mediated adhesion and spreading of human dermal microvascular endothelial cells (HDMVEC). When HDMVEC were cultured in microtiter plate wells coated with concentrations of C1q ranging from 0 to 50 microg/ml, a specific and dose-dependent adhesion and spreading was observed. The extent of adhesion and spreading was similar to the adhesion seen on collagen-coated wells. Spreading (68 +/- 12%) and to a moderate extent adhesion (47 +/- 9%) were inhibited by anti-gC1q-R mAb 60.11. Similar effects were noted with polyclonal anti-cC1q-R but not with control nonimmune IgG. The two Abs had a slight additive effect (75 +/- 13% inhibition) when mixed together in the proportion of 100 microg/ml anti-gC1q-R and 30 microg/ml anti-cC1q-R. More importantly, a 100% inhibition of spreading, but not adhesion, to C1q-coated wells was observed when HDMVEC were cultured in the presence of 30 microM of the peptide GRRGDSP but not GRRGESP. Furthermore, while anti-beta(1) integrin Ab blocked both adhesion and spreading, anti-alpha(5) integrin blocked only spreading and not adhesion. Ag capture ELISA of endothelial cell membrane proteins using polyclonal anti-gC1q-R showed the presence of not only beta(1) and alpha(5) integrins but also CD44. Taken together these results suggest that endothelial cell adhesion and spreading require the cooperation of both C1qRs and beta(1) integrins and possibly other membrane-spanning molecules.
Feng
Xiaodong
X
Department of Dermatology, State University of New York, Health Sciences Center T-16-040, Stony Brook, NY 11794, USA.
Tonnesen
Marcia G
MG
Peerschke
Ellinor I B
EI
Ghebrehiwet
Berhane
B
D000906:Antibodies / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D037282:Calreticulin; D002352:Carrier Proteins; D002448:Cell Adhesion; D002478:Cells, Cultured; D015922:Complement C1q / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D020405:Dermis / Q000098:blood supply / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D018960:Hyaluronan Receptors; D019012:Integrin beta1 / Q000276:immunology / Q000502:physiology*; D016023:Integrins / Q000502:physiology; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D018448:Models, Immunological; D009842:Oligopeptides / Q000494:pharmacology; D011951:Receptors, Complement / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D012261:Ribonucleoproteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*
D000906:Antibodies; C087299:C1QBP protein, human; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D019012:Integrin beta1; D016023:Integrins; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D009842:Oligopeptides; D011951:Receptors, Complement; D012261:Ribonucleoproteins; C032402:complement 1q receptor; C047981:arginyl-glycyl-aspartic acid; D015922:Complement C1q