| 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).
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