title: Toll-like receptor expression and function in the COV434 granulosa cell line.
abstract: OBJECTIVE
  Bacterial infections perturb ovarian follicle function, despite the lack of immune cells such as macrophages within healthy ovarian follicles. This study examined whether the granulosa cells that line ovarian follicles could, like macrophages, use Toll-like receptors (TLRs) to detect pathogen-associated molecular patterns (PAMPs) and initiate inflammation.
  
  
  METHODS
  The COV434 human granulosa and THP-1 macrophage cell lines were used to determine the expression of TLRs and measure the production of cytokines, chemokines and estradiol in response to the PAMPs lipopolysaccharide, peptidoglycan, lipoteichoic acid and flagellin from bacteria.
  
  
  RESULTS
  The THP-1 and granulosa cells expressed mRNA for TLR1-10 and TLR4-10, respectively. The supernatants of THP-1 cells accumulated IL-1β, IL-6, IL-8 and CCL5 in response to PAMPs. Treatment of granulosa cells with PAMPs increased expression of IL1B mRNA after 3 hr, but did not change the accumulation of IL-1β, IL-6, IL-8, CCL5 or estradiol. Granulosa cells produced IL-8 constitutively, and this was reduced using chemical inhibitors for p38 and JNK mitogen-activated protein kinases.
  
  
  CONCLUSIONS
  The COV434 human granulosa cell line expresses TLRs and constitutively secretes IL-8 but only mounts an inflammatory response to PAMPs at the transcriptional level.
authors.affiliation: Institute of Life Science, School of Medicine, Swansea University, UK.
authors.forename: Jennifer Claire
authors.initials: JC
authors.lastname: Price
authors.forename: James
authors.initials: J
authors.lastname: Cronin
authors.forename: Iain Martin
authors.initials: IM
authors.lastname: Sheldon
mesh_terms: D002460:Cell Line; D003470:Culture Media; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D004958:Estradiol / Q000096:biosynthesis / Q000276:immunology; D005260:Female; D005408:Flagellin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D006107:Granulosa Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D010457:Peptidoglycan / Q000494:pharmacology; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000276:immunology; D013682:Teichoic Acids / Q000494:pharmacology; D051193:Toll-Like Receptors / Q000235:genetics* / Q000276:immunology; D014158:Transcription, Genetic / Q000187:drug effects* / Q000276:immunology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D003470:Culture Media; D016207:Cytokines; D008070:Lipopolysaccharides; D010457:Peptidoglycan; D020033:Protein Isoforms; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D013682:Teichoic Acids; D051193:Toll-Like Receptors; D005408:Flagellin; D004958:Estradiol; C009900:lipoteichoic acid; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4