title: Induction of cyclooxygenase (COX)-2 in human vaginal epithelial cells in response to TLR ligands and TNF-α.
abstract: OBJECTIVE
  Mucosal inflammation caused by infections of the female lower genital tract is considered to be an important cofactor for HIV transmission. We hypothesize that COX-2, a key inflammation-related enzyme, is involved in these responses and is upregulated by microbial ligands and pro-inflammatory cytokines.
  
  
  METHODS
  Human vaginal epithelial cells (VK-2/E6E7) and ectocervical biopsy tissues were stimulated with TLR ligands and the cytokine TNF-α, used as surrogates of vaginal infections, and assessed for COX-2 expression and activity by microarray, real-time RT-PCR, immunoblotting, immunohistochemistry, and ELISA.
  
  
  RESULTS
  TLR agonists and TNF-α induce transcriptional and translational expression of COX-2 in vaginal cells. TLR ligands, MALP2, Pam3CSK4, LTA, and imiquimod induced high epithelial COX-2 expression, while zymosan and poly dI:dC induced very low enzyme expression. Induced mRNA and protein expression correlated with increased COX-2 activity, which led to increased levels of PGE(2) in the cell culture supernatant. These cell-based findings were confirmed in primary cervicovaginal tissue explants.
  
  
  CONCLUSIONS
  Induction of COX-2 expression and activity and the consequent increased levels of prostaglandins are common inflammatory pathways in human cervicovaginal epithelial cells and tissues in response to diverse TLR ligands and pro-inflammatory cytokines. These findings are relevant to the understanding of genital mucosal inflammation, its potential treatment, and its possible relationship with increased tissue susceptibility to HIV-1 infection.
authors.affiliation: Department of Obstetrics and Gynecology, CONRAD, Eastern Virginia Medical School, Norfolk, VA, USA.
authors.forename: Theresa
authors.initials: T
authors.lastname: Joseph
authors.forename: Irina A
authors.initials: IA
authors.lastname: Zalenskaya
authors.forename: Nazita
authors.initials: N
authors.lastname: Yousefieh
authors.forename: Suzanne D
authors.initials: SD
authors.lastname: Schriver
authors.forename: Lyn C
authors.initials: LC
authors.lastname: Cote
authors.forename: Neelima
authors.initials: N
authors.lastname: Chandra
authors.forename: Gustavo F
authors.initials: GF
authors.lastname: Doncel
mesh_terms: D000634:Aminoquinolines / Q000494:pharmacology; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000096:biosynthesis* / Q000235:genetics; D015232:Dinoprostone / Q000096:biosynthesis; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D006801:Humans; D000077271:Imiquimod; D008024:Ligands; D055666:Lipopeptides / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D011089:Polydeoxyribonucleotides / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D013682:Teichoic Acids / Q000494:pharmacology; D051193:Toll-Like Receptors / Q000819:agonists / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015854:Up-Regulation; D014621:Vagina / Q000166:cytology / Q000378:metabolism; D015054:Zymosan / Q000494:pharmacology
chemical_list: D000634:Aminoquinolines; D008024:Ligands; D055666:Lipopeptides; D008070:Lipopolysaccharides; C508075:Pam(3)CSK(4) peptide; D011089:Polydeoxyribonucleotides; D012333:RNA, Messenger; D013682:Teichoic Acids; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; C031156:poly d(I-C); C009900:lipoteichoic acid; D015054:Zymosan; C106335:macrophage stimulatory lipopeptide 2; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D015232:Dinoprostone; D000077271:Imiquimod