Antiphospholipid immunoglobulin G antibodies reduce annexin-V levels on syncytiotrophoblast apical membranes and in culture media of placental villi

OBJECTIVES: The mechanism by which antiphospholipid antibodies are associated with pregnancy loss and thromboembolism has not been established. We previously showed that annexin-V, a phospholipid-binding protein with potent anticoagulant activity, is present on the apical membranes or the syncytiotrophoblasts that line placental villi and that this protein is reduced, by immunohistochemistry, on placentas of patients with antiphospholipid antibodies. We therefore investigated whether annexin-V in apical membranes of placental villi is quantitatively reduced by antiphospholipid antibody immunoglobulin G. STUDY DESIGN: Placentas were obtained from an index patient with an antiphospholipid syndrome with intrauterine growth restriction and from a patient with an uncomplicated pregnancy who were both delivered by cesarean section. Apical villous membranes were isolated and annexin-V levels were measured by enzyme-linked immunosorbent assay. We then studied the effects of antiphospholipid immunoglobulin G on placental villous apical annexin-V in vitro. Antiphospholipid immunoglobulin G was isolated from the sera of five different patients with antiphospholipid antibody syndrome along with five paired control immunoglobulin Ga. Short-term cultures were established from normal placental villi and were exposed to the antibodies, after which isolated apical membranes and culture media were immunoassayed for annexin-V levels. RESULTS: Measurements of apical membrane-associated annexin-V from the antiphospholipid placenta showed significantly less apical membrane-associated annexin-V than did the normal placenta (mean +/- SEM: 4.9 +/- 0.4 mu g/gm villi for antiphospholipid placenta vs 10.2 +/- 0.6 mu g/gm villi for control, p < 0.001, n = 4). Exposure of placental villous cultures to five different antiphospholipid immunoglobulin Gs for 24 hours resulted in significant reduction of the levels of apical membrane annexin-V (mean +/- SEM: 3.9 +/- 0.3 mu g/gm villi) compared with paired controls (5.1 +/- 0.3 mu g/gm villi, p = 0.02). Villi incubated with the different antiphospholipid immunoglobulin Gs had significantly less annexin-V in conditioned media (mean +/- SEM: 45.1 +/- 4.9 ng/gm villi) compared with the paired normal immunoglobulin G control levels (72.6 +/- 11.4 ng/gm villi, p = 0.03). CONCLUSIONS: Antiphospholipid immunoglobulin G reduces the levels of syncytiotrophoblast apical membrane-associated annexin-V in placental villi and the release of annexin-V into surrounding media. Reduction of this anticoagulant protein at the maternal-fetal interface may account for the pregnancy loss observed in patients with antiphospholipid syndrome. Short-term culture of placental villi may offer an in vitro model to further study the mechanism of this effect of antiphospholipid antibodies.