PP040. Activation of endosomal toll-like receptors in first trimester trophoblasts

Abstract

Introduction: A mild systemic inflammation may be beneficial to normal pregnancy, however exaggerated inflammation may contribute to pregnancy complications. Infections and cell stress or damage may evoke placental inflammation by activation of Toll-like receptors (TLRs). TLR3, TLR7, TLR8 and TLR9 are located intracellulary on endosomes and are activated by nucleic acids from microbes and damaged cells. Trophoblasts play a crucial role during placentation, and the role of TLRs in first trimester trophoblasts needs to be determined.

Objectives: To characterize endosomal TLR gene expression and activation in first trimester trophoblasts, to extend knowledge of endosomal TLR involvement in placental inflammation.

Methods: Primary trophoblasts were isolated from six first trimester placentas by enzyme degradation and gradient centrifugation. Gene expression of TLR3, TLR7, TLR8 and TLR9 in primary first trimester trophoblasts and the trophoblast cell line BeWo was quantified by RT-qPCR. The trophoblasts were stimulated with ligands for endosomal TLRs, and release of pro-inflammatory cytokines was analyzed by multiplex.

Results: Primary first trimester trophoblasts showed gene expression of all endosomal TLRs, and endosomal TLR activation gave increased production of the pro-inflammatory cytokines IL-6, IL-8, and IP-10.

Conclusion: Primary first trimester trophoblasts express functional endosomal TLRs, indicating TLR-mediated trophoblast involvement in early placental inflammation.