Virginia Research Day 2022

Medical Student Research Biomedical

08 Do Cytokine Profiles In Maternal Mouse Macrophages And Placentas Influence Formation Of Neural Tube Defects In QAC Exposed Mice?

Justin Kula; Dylan Davis; Terry C. Hrubec Corresponding author: jkula@vcom.edu

VCOM Virginia

for NTD. Macrophages will be collected from maternal bone marrow. All embryos from each litter will be examined for the presence of NTDs, and embryologic age staged by number of somites. Monocytes from bone marrow will be differentiated to macrophages, which are then primed with either prostaglandin E2 (for measurement of the alternative pathway) or interferon gamma (for measurement of the canonical pathway) and then challenged with LPS. The supernatants of stimulated cells will be collected, and concentrations of IL-6, TNF- and both pro-inflammatory and anti- inflammatory IL-10 will be determined by ELISA. We expect mice exposed to QACs will have an increase in pro-inflammatory cytokines and a decrease in anti-inflammatory cytokines in both BMDMs and the placenta leading to an increased rate of NTD development.

mice alters the cytokine profile to induce a proinflammatory state both systemically and in the microenvironment of the fetus leading to an increased rate of NTD. To investigate this, the cytokine profile in the placenta and BMDMs isolated from mice exposed to QACs will be analyzed and associated with neural tube development. Mice from a QAC-free colony were used to ensure no prior QAC exposure. Mice were divided into three groups of 12 mice per group. Control mice received no exposure, dosed mice received a single oral dose of QACs on day 7 of gestation, and the third group received ambient exposure to quat disinfectants from normal use of the disinfectant in the mouse room. Exposure to QACs on days 7 and 8 of gestation is the critical exposure window for formation of neural tube defects. We will sacrifice the mice on gestational day 10.5. The uterus will be removed and the embryos separated from the placentas. Placenta explants will be cultured for cytokine analysis and the embryos examined

Neural tube defects (NTD) are a common birth defect, affecting 1-5 in 1,000 births worldwide. Quaternary ammonium compounds (QACs) are a common ingredient in disinfectants and cleaning supplies. In mice, QAC exposure causes NTD and alters cytokine production in near term placentas and in the vaginal mucosa. QACs also increase inflammatory cytokines in an immortalized macrophage cell line. The maternal immune system plays an integral role in development of the fetus and can both cause or prevent birth defects. The placenta represents maternal-fetal interface and analysis of cytokines at this level will allow us to determine if QACs cause a proinflammatory state in fetal environment. Bone marrow derived macrophages (BMDMs) represent the maternal systemic immune response to QAC exposure. This study will examine the role of maternal systemic inflammation, and of inflammation at the maternal fetal interface in the production of QAC induced NTDs. We hypothesize that exposure to QACs in pregnant

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