AT(1) receptor agonistic antibodies from preeclamptic patients cause vascular cells to express tissue factor

Background-We recently described autoantibodies (angiotensin-1 receptor aut oantibodies, AT(1)-AA) directed at the AT(1) receptor in the serum of preec lamptic patients, whose placentas are commonly infarcted and express tissue factor (TF). Mechanisms of how AT(1)-AA might contribute to preeclampsia a re unknown. We tested the hypothesis that AT(1)-AA cause vascular smooth mu scle cells (VSMC) to express TF. Methods and Results-IgG from preeclamptic patients containing AT(1)-AA was purified with anti-human IgG columns. AT(1)-AA were separated from the IgG by ammonium sulfate precipitation. We transfected Chinese hamster ovary cel ls overexpressing the AT(1) receptor with TF promoter constructs coupled to a luciferase reporter gene. VSMC were obtained from human coronary arterie s, Extracellular signal-related kinase activation was detected by an in-gel kinase assay. AP-1 activation was determined by electromobility shift assa y. TF was measured by ELISA and detected by immunohistochemistry. Placentas from preeclamptic women stained strongly for TF, whereas control placentas showed far less staining. We proved AT(1)-AA specificity by coimmunoprecip itating the AT(1) receptor with AT(1)-AA but not with nonspecific IgG. Angi otensin (Ang) II and AT(1)-AA both activated extracellular signal-related k inase, AP-1, and the TF promoter transfected VSMC and Chinese hamster ovary cells, but only when the AP-1 binding site was present. We then demonstrat ed TF expression in VSMC exposed to either Ang II or AT(1)-AA. All these ef fects were blocked by losartan. Nonspecific IgG or IgG from nonpreeclamptic pregnant women had a negligible effect. Conclusions-We conclude that AT(1)-AA and Ang II both stimulate the AT(1) r eceptor and initiate a signaling cascade resulting in TF expression. These results show an action of AT(1)-AA on human cells that could contribute to the pathogenesis of preeclampsia.