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Infection and Immunity, May 1999, p. 2371-2376, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Pseudomonas aeruginosa Hemolytic Phospholipase C Suppresses Neutrophil Respiratory Burst Activity

Lance S. Terada,* Kristine A. Johansen, Sogol Nowbar, Adriana I. Vasil, and Michael L. Vasil

University of Colorado Health Sciences Center, Denver, Colorado 80262

Received 3 December 1998/Returned for modification 21 January 1999/Accepted 24 February 1999

Pseudomonas aeruginosa is a persistent pathogen in the airways of patients with cystic fibrosis or bronchiectasis from other causes and appears to have evolved strategies to survive the inflammatory response of the host. We hypothesized that the secreted hemolytic phospholipase C (PLC) of P. aeruginosa (PlcHR) would decrease neutrophil respiratory burst activity. We found that while intact wild-type P. aeruginosa cells stimulated moderate respiratory burst activity from human neutrophils, an isogenic mutant pseudomonas (Delta HR strain) containing a targeted deletion of the plcHR operon induced a much more robust oxidative burst from neutrophils. In contrast, a second pseudomonas mutant (Delta N) containing a disruption in the gene encoding the nonhemolytic PLC (PlcN) was not different from the wild type in stimulating neutrophil O2·- production. Readdition of purified PlcHR to the Delta HR strain suppressed neutrophil O2·- production to levels stimulated by wild-type bacteria. Interestingly, purified PlcHR decreased phorbol myristate acetate (PMA)- but not formyl methionyl-leucyl-proline (fMLP)-induced respiratory burst activity, suggesting interference by PlcHR with a protein kinase C (PKC)-specific signaling pathway. Accordingly, the PKC inhibitor bisindolylmaleimide inhibited the oxidative burst induced by either PMA or intact pseudomonas, but not by fMLP, whereas the p38 kinase inhibitor SB-203580 fully inhibited the respiratory burst induced by fMLP or the PlcHR-replete wild-type bacteria, but not PMA or the PlcHR-deficient Delta HR bacterial mutant. We conclude that expression of PlcHR by P. aeruginosa suppresses bacterium-induced neutrophil respiratory burst by interfering with a PKC-dependent, non-p38 kinase-dependent pathway.

* Corresponding author. Present address: Dallas VAMC and UT Southwestern, Pulmonary and Critical Care (111F), 4500 S. Lancaster Rd., Dallas, TX 75216. Phone: (214) 857-1475. Fax: (214) 857-0340. E-mail: lterada{at}aol.com.

Infection and Immunity, May 1999, p. 2371-2376, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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