W. Hell et al., CLEAVAGE OF TUMOR-NECROSIS-FACTOR-ALPHA BY LEGIONELLA EXOPROTEASE, APMIS. Acta pathologica, microbiologica et immunologica Scandinavica, 101(2), 1993, pp. 120-126
The role of the major secretory protein of Legionella pneumophila, a z
inc protease, in Legionella infection is not known. Since an important
step of the host reaction in Legionnaires' disease is the production
of tumor necrosis factor-alpha (TNF-alpha) by alveolar macrophages, we
studied the interaction of Legionella protease and U-937 cells with r
espect to TNF-alpha. The Legionella protease was purified by fractiona
ted precipitation, gel filtration and hydrophobic interaction chromato
graphy. The purified enzyme was added to U-937 cells, a promyelocytic
cell line. In the supernatants of PMA-treated U-937 cells we found low
concentrations of TNF-alpha after incubation with protease. Therefore
we pursued the hypothesis of direct enzymatic degradation of TNF-alph
a by Legionella protease. Enzymatic cleavage of TNF-alpha was proven b
y SDS-PAGE, ELISA and TNF-alpha bioassay with L-929 cells. The degrada
tion of TNF-alpha by the Legionella protease was shown in all three sy
stems. Enzymatic degradation of TNF-alpha might be important for the p
athogenesis of Legionnaires' disease.