Proteolytic enzymes and amylase induce cytokine production in human peripheral blood mononuclear cells in vitro

Cancer Biother. Fall 1994;9(3):253-63. doi: 10.1089/cbr.1994.9.253.


In vitro treatment of human peripheral blood mononuclear cells (PBMNC) with proteolytic enzymes (bromelain, papain) and amylase leads to the production of large amounts of tumor necrosis factor-alpha (TNF-alpha), interleukin-1-beta (IL-1 beta), and interleukin-6 (IL-6) in a time and dose dependent manner. Increased TNF-alpha and IL-6 production was already found after 4-6 hours of incubation, and plateau levels were reached after 12-16 hours. Plateau levels up to 1500 pg TNF-alpha/ml/10(6) PBMNC, 13000 pg IL-1 beta/ml/10(6) PBMNC, and 23000 pg IL-6/ml/10(6) PBMNC were observed. Control cultures contained below 35 pg/ml/10(6) PBMNC of TNF-alpha, IL-1 beta or IL-6. In contrast to TNF-alpha which was undetectable after more than 24 hours, peak levels of IL-1 beta and IL-6 were still present at 24 hours. After incubation of the enzyme solution for some hours at 56 degrees C the cytokine inducing capacity disappeared. Neutralization experiments with inactivating antibodies, radioimmunoassay, and western blotting after electrophoretic separation showed that the TNF-like activity found in the lytic assay was due to TNF-alpha. Interferon-alpha (IFN-alpha) and Interferon-gamma (IFN-gamma), which had no effect alone, synergistically increased TNF-alpha production when applied together with the enzymes. A commercial mixture of these enzymes (Wobenzym), which was also investigated, showed a similar concentration and time dependence, as well as synergism with the interferons. A synergistic effect on TNF-alpha production was also found with the enzymes and phorbol ester (PMA).

MeSH terms

  • Amylases / pharmacology*
  • Bromelains / pharmacology*
  • Cells, Cultured
  • Cytokines / biosynthesis
  • Cytokines / genetics
  • Drug Synergism
  • Gene Expression Regulation / drug effects
  • Humans
  • Interferons / biosynthesis
  • Interferons / genetics
  • Interleukin-1 / biosynthesis*
  • Interleukin-1 / genetics
  • Interleukin-6 / biosynthesis*
  • Interleukin-6 / genetics
  • Leukocytes, Mononuclear / drug effects*
  • Leukocytes, Mononuclear / metabolism
  • Papain / pharmacology*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tumor Necrosis Factor-alpha / biosynthesis*
  • Tumor Necrosis Factor-alpha / genetics


  • Cytokines
  • Interleukin-1
  • Interleukin-6
  • Tumor Necrosis Factor-alpha
  • Bromelains
  • Interferons
  • Amylases
  • Papain
  • Tetradecanoylphorbol Acetate