Villous explants from preeclamptic placentas induce sFlt1 in PBMCs: An ex vivo co-culture study

Abstract

Objectives: Soluble Flt1 (sFlt1) is an anti-angiogenic protein linked to the pathology of preeclampsia (PE). While the placenta serves as the major organ producing sFlt1 during normal pregnancy, peripheral blood mononuclear cells (PBMCs), endothelial cells, and stromal cells also produce sFlt1. The key question is 'what drives the overexpression of sFlt1 observed during PE?' In the present work we show evidence for sFlt1 over-expression in PBMCs due to interaction with placental villi from PE patients.

Study design: sFlt1 production by PBMCs is estimated by using two blood collection methods with different coagulation chemistry. PBMCs were then cultured with homologous villous explants and heterologous villous explants to determine the effects of the interaction between the two tissues.

Main outcome measures: sFlt1 levels were estimated using real time PCR, ELISA, and gel electrophoresis.

Results: Plasma samples obtained using CTAD as anti-coagulant showed 16-23% less sFlt1 compared to plasma collected in EDTA. Preeclamptic PBMCs showed higher basal level of sFlt1 mRNA. In addition, we show evidence of placental interaction as a cause of sFlt1 overexpression in PBMCs using homologous and heterologous co-culture system. However, during co-culture, we observed that while the sFlt1 expression in PE PBMCs is increased, PE villous explants show reduced sFlt1 RNA expression.

Conclusion: sFlt1 was produced in significant amounts by preeclamptic PBMCs, and ex vivo studies show that the placenta induces this over-expression. In contrast, exposure to PBMCs appears to decrease sFlt1 production by preeclamptic placenta.

Keywords: Co-culture; PBMCs; Placental villous explants; Preeclampsia; sFlt1.