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Bactericidal properties of group IIa secreted phospholipase A₂ against Pseudomonas aeruginosa clinical isolates

¹INSERM CPTP-U563, CHU Purpan, 1 Place du Dr Baylac, TSA 40031, 31059 Toulouse Cedex 9, France ^2,4Laboratoire de Bactériologie-Hygiène² and INSERM U583⁴, CHU Rangueil, 1 Avenue Jean Poulhes, TSA 50032, 31059 Toulouse Cedex 9, France ³Laboratoire de Microbiologie, Faculté de Pharmacie, 27 Chemin des Maraîchers, 31400 Toulouse, France

Correspondence Anne Dubouix dubouix.a{at}chu-toulouse.fr

Received April 25, 2003
Accepted August 11, 2003

It has been shown that human group IIa secreted phospholipase A₂ (sPLA₂), found at high levels in inflammatory fluids, displays direct bactericidal properties against Gram-positive bacteria, while activity against Gram-negative bacteria requires the complement system or additional co-factors produced by neutrophils. Pseudomonas aeruginosa, an increasingly prevalent opportunistic human pathogen, is the most common Gram-negative rod found in cystic fibrosis lung infections, where it is associated with an inflammatory environment. Because murine intestinal group II sPLA₂ produced by Paneth cells has been shown to be directly bactericidal against Gram-negative bacteria, IIa sPLA₂ activity against P. aeruginosa clinical isolates was evaluated and provides the first evidence that the enzyme can be fully bactericidal in a concentration- and time-dependent manner against Gram-negative rods. Furthermore, it was demonstrated that these bactericidal properties were unaffected by high protein and salt concentrations, as observed in cystic fibrosis secretions, and that bacterial killing paralleled phospholipid hydrolysis. Finally, no cytotoxicity was observed when IIa sPLA₂ was incubated with human pulmonary cells, highlighting its potential use to synergize bactericidal antibiotics by promoting sublethal alterations of the bacterial cell wall.

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