title: [The activation of TRIF-dependent signaling pathway in THP-1 cells induced by β₂ GPI/anti-β₂ GPI antibodies complex].
abstract: OBJECTIVE
  To observe whether the TIR-domain-containing adaptor inducing interferon-β (TRIF) is activated in THP-1 cells treated with β₂ GPI/anti-β₂ GPI complex and investigate the roles of TRIF-dependent signaling pathway of Toll-like receptor 4 (TLR4) in antiphospholipid syndrome (APS).
  
  
  METHODS
  The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with β₂ GPI/anti-β₂ GPI complex. And the level of TRIF mRNA in THP-1 cells was detected by Real-time PCR (RT-PCR), TRIF protein expression was investigated by western blotting, respectively. Furthermore, whether TLR4 inhibitor, TAK-242, could interrupt the expression of TRIF as well as some inflammatory cytokines such as IL-6, IL-8 and TNF-α in THP-1 cells stimulated with β₂ GPI/anti-β₂ GPI complex was also investigated.
  
  
  RESULTS
  Both mRNA and protein levels of TRIF could be significantly increased in THP-1 cells with treatment of β₂ GPI/anti-β₂ GPI complex (100 mg/L). The expression of TRIF was shown in a manner of time-dependence, with the maximal levels at 1 hour (mRNA) and 2 hour (protein) stimulation respectively. The β₂ GPI/anti-β₂ GPI complex-induced TRIF and inflammatory cytokines including IL-6, IL-8 and TNF-α expression in THP-1 cells could be inhibited by TAK-242 (5 μmol/L).
  
  
  CONCLUSIONS
  TRIF-dependent signaling pathway of Toll-like receptor 4 is involved in the activation of THP-1 cells induced by β₂ GPI/anti-β₂GPI complex, suggesting that TRIF may play an important role in the pathogenesis of antiphospholipid syndrome.
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China. xiehongxian007@163.com
authors.forename: Hong-xiang
authors.initials: HX
authors.lastname: Xie
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.forename: Hai-bo
authors.initials: HB
authors.lastname: Wang
authors.forename: Dong-dong
authors.initials: DD
authors.lastname: Chen
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Xian-mei
authors.initials: XM
authors.lastname: Zhang
authors.forename: Long-fei
authors.initials: LF
authors.lastname: Xia
authors.forename: Yuan
authors.initials: Y
authors.lastname: Mu
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000502:physiology*; D000936:Antigen-Antibody Complex / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics; D016209:Interleukin-8 / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D015398:Signal Transduction / Q000502:physiology*; D051197:Toll-Like Receptor 4 / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D053482:beta 2-Glycoprotein I / Q000276:immunology*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D000936:Antigen-Antibody Complex; D015850:Interleukin-6; D016209:Interleukin-8; D012333:RNA, Messenger; C470409:TICAM1 protein, human; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D053482:beta 2-Glycoprotein I