Stimulation of the secretion of latent cysteine proteinase activity by tumor necrosis factor alpha and interleukin-1

Arthritis Rheum. 1993 Jun;36(6):772-80. doi: 10.1002/art.1780360606.


Objective: Cultured synovial fibroblast-like cells from 3 patients with rheumatoid arthritis (RA) and 3 patients with osteoarthritis (OA) were evaluated for their potential to secrete cysteine proteinases spontaneously and after stimulation by tumor necrosis factor alpha (TNF alpha) or interleukin-1 (IL-1).

Methods: Culture media and cell lysates were analyzed before and after high performance liquid chromatography (HPLC) using the enzymatic substrate, Z-Phe-Arg-AMC, and by immunoblotting with anti-cathepsin B antiserum. Immunolocalization of cathepsin B was studied on cell monolayers.

Results: Latent cysteine proteinase activity was found to be secreted spontaneously by cultured synovial fibroblast-like cells. This activity was increased after treatment with either TNF alpha or IL-1. Stimulated protease activity was eluted by HPLC at a peak coincident with that of purified cathepsin B. By immunoblot, cell supernatants contained a 43-kd form of cathepsin B, while cell lysates contained a 30-kd form, consistent, respectively, with cathepsin B before and after cleavage of its propeptide. An intracellular increase in cathepsin B after treatment with TNF alpha was also seen with immunohistochemical studies.

Conclusion: TNF alpha (in the 6 cases studied) and IL-1 (in 4 cases) stimulated the secretion of a latent cysteine proteinase activity from synovial fibroblast-like cells, which appears to represent primarily cathepsin B.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arthritis, Rheumatoid / pathology
  • Cathepsin B / chemistry
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Cysteine Endopeptidases / metabolism*
  • Enzyme Activation / drug effects
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Interleukin-1 / pharmacology*
  • Osteoarthritis / pathology
  • Synovial Membrane / cytology
  • Synovial Membrane / enzymology
  • Tumor Necrosis Factor-alpha / pharmacology*


  • Interleukin-1
  • Tumor Necrosis Factor-alpha
  • Cysteine Endopeptidases
  • Cathepsin B