Maternal autoantibodies from preeclamptic patients activate angiotensin receptors on human trophoblast cells

Abstract

Recent evidence indicates that preeclampsia is associated with the presence of autoantibodies capable of activating the angiotensin II receptor, AT1. We sought to evaluate the role of AT1 agonistic autoantibodies (AT1-AA) in two major features of preeclampsia-increased plasminogen activator inhibitor-1 (PAI-1) production and shallow trophoblast invasion. This study included 38 pregnant patients, 20 of whom had severe preeclampsia and 18 normotensive individuals. Immunoglobulin (Ig)G was purified from these individuals, and the presence of AT1-AA was determined based on its ability to stimulate an increase in the contraction rate of cultured rat neonatal cardiac myocytes. Immortalized human trophoblasts were chosen to study PAI-1 production and secretion after treatment with IgG from normotensive and preeclamptic women. An in vitro Matrigel invasion assay was used to test the effect of AT1-AA on the invasive properties of human trophoblasts. Losartan and cyclosporin A were used to determine whether the AT1-AA-induced stimulation of PAI-1 secretion is through the AT1 receptor and the calcineurin-nuclear factor of activated t-cells (NFAT)-dependent pathway. The results show that IgG from 18 of 20 severely preeclamptic women stimulated increased cardiomyocyte contraction rates of 20-40 beats per minute. A significant stimulation of PAI-1 secretion from human trophoblasts was observed with IgG from the same 18 of 20 patients with severe preeclampsia. Of IgG obtained from 18 normotensive pregnant patients, only two showed a relatively low level of biologic activity in the cardiomyocyte contraction and PAI-1 secretion assays. Activation of AT1 receptors by AT1-AA was blocked by losartan (an AT1 receptor antagonist) and by a seven amino acid peptide corresponding to a sequence present on the second extracellular loop of the AT1 receptor. Activation of AT1 receptors by AT1-AA resulted in decreased trophoblast invasiveness as determined by the in vitro Matrigel invasion assay. Additional data indicate that AT1 receptor activation by AT1-AA is followed by the downstream activation of the calcium-dependent calcineurin-NFAT signaling pathway leading to increased PAI-1 gene expression. Our findings suggest that maternal autoantibody with the ability to activate AT1 receptors may account for two features of preeclampsia, increased PAI-1 production and shallow trophoblast invasion.