title: Human granulosa-luteal cells initiate an innate immune response to pathogen-associated molecules.
abstract: The microenvironment of the ovarian follicle is key to the developmental success of the oocyte. Minor changes within the follicular microenvironment can significantly disrupt oocyte development, compromising the formation of competent embryos and reducing fertility. Previously described as a sterile environment, the ovarian follicle of women has been shown to contain colonizing bacterial strains, whereas in domestic species, pathogen-associated molecules are concentrated in the follicular fluid of animals with uterine infection. The aim of this study is to determine whether human granulosa-luteal cells mount an innate immune response to pathogen-associated molecules, potentially disrupting the microenvironment of the ovarian follicle. Human granulosa-luteal cells were collected from patients undergoing assisted reproduction. Cells were cultured in the presence of pathogen-associated molecules (LPS, FSL-1 and Pam3CSK4) for 24h. Supernatants and total RNA were collected for assessment by PCR and ELISA. Granulosa-luteal cells were shown to express the molecular machinery required to respond to a range of pathogen-associated molecules. Expression of TLR4 varied up to 15-fold between individual patients. Granulosa-luteal cells increased the expression of the inflammatory mediators IL1B, IL6 and CXCL8 in the presence of the TLR4 agonist E. coli LPS. Similarly, the TLR2/6 ligand, FSL-1, increased the expression of IL6 and CXCL8. Although no detectable changes in CYP19A1 or STAR expression were observed in granulosa-luteal cells following challenge, a significant reduction in progesterone secretion was measured after treatment with FSL-1. These findings demonstrate the ability of human granulosa-luteal cells to respond to pathogen-associated molecules and generate an innate immune response.
authors: D. H. Barron Reproductive and Perinatal Biology Research ProgramGainesville, Florida, USA Department of Animal SciencesUniversity of Florida, Gainesville, Florida, USA.
authors: Laila A
authors: LA
authors: Ibrahim
authors: D. H. Barron Reproductive and Perinatal Biology Research ProgramGainesville, Florida, USA Department of Obstetrics and GynecologyCollege of Medicine, University of Florida, Gainesville, Florida, USA.
authors: Joseph M
authors: JM
authors: Kramer
authors: D. H. Barron Reproductive and Perinatal Biology Research ProgramGainesville, Florida, USA Department of Obstetrics and GynecologyCollege of Medicine, University of Florida, Gainesville, Florida, USA.
authors: R Stan
authors: RS
authors: Williams
authors: D. H. Barron Reproductive and Perinatal Biology Research ProgramGainesville, Florida, USA Department of Animal SciencesUniversity of Florida, Gainesville, Florida, USA jbromfield@ufl.edu.
authors: John J
authors: JJ
authors: Bromfield
mesh_terms: D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology*; D005260:Female; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects / Q000276:immunology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008184:Luteal Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D009842:Oligopeptides / Q000494:pharmacology*; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000819:agonists
chemical_list: D016207:Cytokines; D004075:Diglycerides; C460864:FSL-1 lipoprotein, synthetic; D008070:Lipopolysaccharides; D009842:Oligopeptides; D051197:Toll-Like Receptor 4