IDENTIFICATION OF CLQ AS THE HEAT-LABILE SERUM COFACTOR REQUIRED FOR IMMUNE-COMPLEXES TO STIMULATE ENDOTHELIAL EXPRESSION OF THE ADHESION MOLECULES E-SELECTIN AND INTERCELLULAR AND VASCULAR CELL-ADHESION MOLECULES-1

To examine the role of complement components as regulators of the expression of endothelial adhesive molecules in response to immune complexes (ICs), we determined whether ICs stimulate both endothelial adhesiveness for leukocytes and expression of E-selectin and intercellular and vascular cell adhesion molecules 1 (ICAM-1 and VCAM-1), We found that ICs [bovine serum albumin (BSA)-anti-BSA] stimulated endothelial cell adhesiveness for added leukocytes in the presence of complement-sufficient normal human serum (NHS) but not in the presence of heat-inactivated serum (HIS) or in tissue culture medium alone. Depletion of complement component C3 or C8 from serum did not prevent enhanced endothelial adhesiveness stimulated by ICs, In contrast, depletion of complement component Clq markedly inhibited IC-stimulated endothelial adhesiveness for leukocytes, When the heat-labile complement component Clq was added to HIS, the capacity of ICs to stimulate endothelial adhesiveness for leukocytes was completely restored, Further evidence for the possible role of Clq in mediating the effect of ICs on endothelial cells was the discovery of the presence of the 100- to 126-kDa Clq-binding protein on the surface of endothelial cells (by cytofluorography) and of message for the 33-kDa Clq receptor in resting endothelial cells (by reverse transcription-PCR), Inhibition of protein synthesis by cycloheximide blocked endothelial adhesiveness for leukocytes stimulated by either interleukin 1 or ICs in the presence of NHS. After stimulation with ICs in the presence of NHS, endothelial cells expressed increased numbers of adhesion molecules (E-selectin, ICAM-1, and VCAM-1), Endothelial expression of adhesion molecules mediated, at least in part, endothelial adhesiveness for leukocytes, since leukocyte adhesion was blocked by monoclonal antibodies directed against E-selectin, These studies show that ICs stimulate endothelial cells to express adhesive proteins for leukocytes in the presence of a heat-labile serum factor, That factor appears to be C1q.