NOD Protein Expression and Function in First Trimester Trophoblast Cells

Abstract

Through the expression of pattern recognition receptors, the trophoblast can recognize and respond to infectious microorganisms and, therefore, participate in the control of pathogens that may compromise fetal well-being. We hypothesize that the trophoblast has the ability to sense invasive intracellular bacteria through the cytoplasmic-based nucleotide-binding oligomerization domain (NOD) proteins. The aim of this study was to characterize the expression and function of NOD proteins in first trimester trophoblast cells. NOD1 and NOD2 expressions by first trimester trophoblast cells were evaluated by immunohistochemistry, Western blot analysis and reverse transcription-polymerase chain reaction. The effect of NOD2 activation on trophoblast cells was determined by analyzing the cytokine response following treatment with muramyl dipeptide (MDP). Both NOD1 and NOD2 were expressed by first trimester placental villi and localized to trophoblast cells. Moreover, NOD1, NOD2 and the signaling effector protein, RIP-like interacting CLARP kinase (RICK), were all expressed by isolated trophoblast cells. Following exposure to the NOD2 ligand, MDP, trophoblast cells generated a pro-inflammatory cytokine response. This response was confirmed to be specific, as an NOD2-deficient trophoblast cell line failed to respond to MDP unless transfected with NOD2. These findings suggest that, through the expression and function of NOD proteins, first trimester trophoblast cells are able to recognize and respond to invasive intracellular pathogens that may have evaded other forms of pattern recognition.