[8, 9] The compound PGE2 is an arachidonic acid-derived lipid mediator generated in abundance at sites of infection and inflammation as a result of the rapid up-regulation of cyclooxygenase-2 and microsomal PGE synthase-1 enzymes.[10] It is also an important hormonal regulator of reproduction that is generated in the uterus where it is involved in early and late processes ranging from implantation of the fertilized egg to parturition.[11]

PGE2 is a highly potent modulator of innate and adaptive immunity that influences cell behavior through the ligation of its four distinct G-protein-coupled E-prostanoid (EP) receptors, numbered EP1-4.[12, 13] Both EP2 and EP4 are potent immunoregulatory receptors that share the capacity to increase intracellular concentrations of cyclic adenosine monophosphate (cAMP) within seconds to minutes of PGE2 binding.[13, 14] PGE2-dependent increases in cAMP have been shown to impair the phagocytic ability of different macrophage MAPK Inhibitor Library manufacturer types against a range of pathogens,[15-18] and it can be suggested that such effects might have evolved to limit the extent of host inflammatory responses or trigger the resolution of inflammation. However, in clinical situations such as pregnancy and the puerperium, where local and systemic PGE2 levels are elevated for physiological reasons,[19-21] the immunosuppressive effects of PGE2 might be maladaptive, particularly when an opportunistic this website pathogen such as C. sordellii gains access

to the normally uninfected uterus (or surrounding soft tissue). The purpose of this study was to address the question of whether PGE2 and cAMP-signaling cascades could regulate the phagocytosis of C. sordellii by human macrophages

and to determine the involvement and Amine dehydrogenase relative importance of EP2 and EP4 receptors in such regulation. A better understanding of endogenous regulators of innate immunity will enhance efforts to develop better preventive and therapeutic options against reproductive tract infections. Phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 cells (a human macrophage-like cell line) were used in this study. These cells were obtained from the American Type Culture Collection (ATCC, TIB-202; Manassas, VA, USA) and cultured in RPMI 1640 (Invitrogen, Carlsbad, CA, USA) supplemented with 1% antibiotic-antimycotic (Invitrogen) and 10% charcoal-/dextran-treated fetal bovine serum (FBS; HyClone, Waltham, MA, USA), referred to as RPMI +/+. Cells were passaged every 2–4 days and were used through the 10th passage, at which time a new culture was started. THP-1 cells were matured into macrophages by culturing with 100 nm PMA (Sigma-Aldrich, St. Louis, MO, USA) in RPMI +/+ for 24 hr at 37°C with 5% CO2. Cells were detached from the flask with non-enzymatic cell dissociation solution (Sigma-Aldrich) and gentle scraping. Phorbol-12-myristate-13-acetate-activated THP-1 cells were used for all experiments presented here, unless otherwise noted.