Effect of Pseudomonas aeruginosa exotoxin A on endotoxin-induced tumour necrosis factor production in murine lung

The ability of several Pseudomonas aeruginosa exo-enzymes, including exotox in A (ETA), to induce inflammation and their influence on endotoxin-induced tumour necrosis factor (TNF) production in murine lung were evaluated. Int ratracheal administration of lipopolysaccharide (LPS; 0.1-10 mu g/mouse), 2 (-1) LD50 of P. aeruginosa alkaline protease (7.5 mu g/mouse) and elastase (1.2 mu g/mouse) elevated total cell number and the percentage of neutrophi ls in broncho-alveolar lavage fluid (BALF), whereas ETA (0.1 mu g/mouse) di d not. LPS induced TNF production in BALF in a dose-dependent manner, where as the P. aeruginosa exo-enzymes did not, When ETA was inoculated into the respiratory tract before LPS, production of TNF in BALF was significantly s uppressed in a dose-dependent manner. ETA also suppressed TNF production by alveolar macrophages (AMs) stimulated with LPS in vitro, Flow cytometric a nalysis showed that ETA markedly reduced the expression of CD14 and CD11c/C D18 on the surface of AMs, ETA also depressed partially the expression of T NF-alpha mRNA in AMs, These findings suggest that ETA regulates TNF product ion in murine lung by suppressing LPS receptor expression, mRNA expression and protein synthesis and/or secretion of TNF.