Rheumatoid arthritis synovial macrophages express the Fas-associated death domain-like interleukin-1beta-converting enzyme-inhibitory protein and are refractory to Fas-mediated apoptosis

Arthritis Rheum. 2001 Jan;44(1):21-30. doi: 10.1002/1529-0131(200101)44:1<21::AID-ANR4>3.0.CO;2-8.


Objective: The chronic inflammation and progressive joint destruction observed in rheumatoid arthritis (RA) are mediated in part by macrophages. A paucity of apoptosis has been observed in RA synovial tissues, yet the mechanism remains unknown. The present study sought to characterize the expression of Fas, Fas ligand (FasL), and Fas-associated death domain-like interleukin-1beta-converting enzyme-inhibitory protein (FLIP), and to quantify the apoptosis induced by agonistic anti-Fas antibody, using mononuclear cells (MNC) isolated from the peripheral blood (PB) and synovial fluid (SF) of RA patients.

Methods: The expression of Fas, FasL, and FLIP and apoptosis induced by agonistic anti-Fas antibody in MNC from the PB and SF of RA patients were determined by flow cytometry. Immunohistochemistry employing a monospecific anti-FLIP antibody was performed on RA and osteoarthritis (OA) synovial tissue.

Results: CD14-positive monocyte/macrophages from normal and RA PB and from RA SF expressed equivalent levels of Fas and FasL. Furthermore, unlike the CD14-positive PB monocytes, RA SF monocyte/macrophages were resistant to the addition of agonistic anti-Fas antibody. In contrast, both CD14-positive PB and SF monocyte/macrophages were sensitive to apoptosis mediated by a phosphatidylinositol 3-kinase inhibitor. Intracellular staining of the caspase 8 inhibitor, FLIP, in CD14-positive SF monocyte/macrophages revealed a significant up-regulation of FLIP compared with normal and RA PB monocytes. Immunohistochemical analysis of synovial tissue from RA and OA patients revealed increased FLIP expression in the RA synovial lining compared with the OA synovial lining. Furthermore, FLIP expression was observed in the CD68positive population in the RA synovial lining. Forced reduction of FLIP by a chemical inhibitor resulted in RA SF macrophage apoptosis that was enhanced by agonistic anti-Fas antibody, indicating that FLIP is necessary for SF macrophage survival.

Conclusion: These data suggest that up-regulation of FLIP in RA macrophages may account for their persistence in the disease. Thus, the targeted suppression of FLIP may be a potential therapeutic strategy for the amelioration of RA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Apoptosis / drug effects
  • Arthritis, Rheumatoid / pathology*
  • Carrier Proteins / analysis*
  • Caspase 8
  • Caspase 9
  • Caspases / biosynthesis
  • Cysteine Proteinase Inhibitors / biosynthesis
  • Fas-Associated Death Domain Protein
  • Fluorescent Antibody Technique
  • Macrophages / chemistry*
  • Monocytes / cytology
  • Serpins / biosynthesis*
  • Synovial Membrane / cytology
  • Viral Proteins*
  • fas Receptor / analysis


  • Adaptor Proteins, Signal Transducing
  • Carrier Proteins
  • Cysteine Proteinase Inhibitors
  • Fas-Associated Death Domain Protein
  • Serpins
  • Viral Proteins
  • fas Receptor
  • interleukin-1beta-converting enzyme inhibitor
  • Caspase 8
  • Caspase 9
  • Caspases