title: AT(1) receptor agonistic antibodies from preeclamptic patients cause vascular cells to express tissue factor.
abstract: BACKGROUND
  We recently described autoantibodies (angiotensin-1 receptor autoantibodies, AT(1)-AA) directed at the AT(1) receptor in the serum of preeclamptic patients, whose placentas are commonly infarcted and express tissue factor (TF). Mechanisms of how AT(1)-AA might contribute to preeclampsia are unknown. We tested the hypothesis that AT(1)-AA cause vascular smooth muscle cells (VSMC) to express TF.
  
  
  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 cells overexpressing the AT(1) receptor with TF promoter constructs coupled to a luciferase reporter gene. VSMC were obtained from human coronary arteries. Extracellular signal-related kinase activation was detected by an in-gel kinase assay. AP-1 activation was determined by electromobility shift assay. 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 coimmunoprecipitating the AT(1) receptor with AT(1)-AA but not with nonspecific IgG. Angiotensin (Ang) II and AT(1)-AA both activated extracellular signal-related kinase, 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 demonstrated TF expression in VSMC exposed to either Ang II or AT(1)-AA. All these effects 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) receptor 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.
authors.affiliation: Franz Volhard Clinic and Max Delbrück Center for Molecular Medicine, Medical Faculty of the Charité, Humboldt University of Berlin, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Dechend
authors.forename: V
authors.initials: V
authors.lastname: Homuth
authors.forename: G
authors.initials: G
authors.lastname: Wallukat
authors.forename: J
authors.initials: J
authors.lastname: Kreuzer
authors.forename: J K
authors.initials: JK
authors.lastname: Park
authors.forename: J
authors.initials: J
authors.lastname: Theuer
authors.forename: A
authors.initials: A
authors.lastname: Juepner
authors.forename: D C
authors.initials: DC
authors.lastname: Gulba
authors.forename: N
authors.initials: N
authors.lastname: Mackman
authors.forename: H
authors.initials: H
authors.lastname: Haller
authors.forename: F C
authors.initials: FC
authors.lastname: Luft
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology; D057911:Angiotensin Receptor Antagonists; D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D016466:CHO Cells; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006224:Cricetinae; D004789:Enzyme Activation; D005260:Female; D006801:Humans; D019808:Losartan / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010920:Placenta / Q000378:metabolism; D011225:Pre-Eclampsia / Q000276:immunology* / Q000378:metabolism; D011247:Pregnancy; D044140:Receptor, Angiotensin, Type 1; D044139:Receptor, Angiotensin, Type 2; D011945:Receptors, Angiotensin / Q000819:agonists* / Q000276:immunology*; D012016:Reference Values; D013925:Thromboplastin / Q000235:genetics / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000502:physiology; D014162:Transfection
chemical_list: D057911:Angiotensin Receptor Antagonists; D000906:Antibodies; D044140:Receptor, Angiotensin, Type 1; D044139:Receptor, Angiotensin, Type 2; D011945:Receptors, Angiotensin; D018808:Transcription Factor AP-1; D000804:Angiotensin II; D013925:Thromboplastin; D020928:Mitogen-Activated Protein Kinases; D019808:Losartan