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Infection and Immunity, March 1999, p. 1125-1130, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Role of Listeria monocytogenes Exotoxins Listeriolysin and Phosphatidylinositol-Specific Phospholipase C in Activation of Human Neutrophils

Ulf Sibelius,1 Eva-Cathrin Schulz,1 Frank Rose,1 Katja Hattar,1 Thomas Jacobs,2 Siegfried Weiss,2 Trinad Chakraborty,3 Werner Seeger,1 and Friedrich Grimminger1,*

Department of Internal Medicine1 and Institute of Medical Microbiology,3 Justus-Liebig-University, Giessen, and Division of Cell Biology and Immunology, National Center for Biotechnology, Braunschweig,2 Germany

Received 20 August 1998/Returned for modification 2 October 1998/Accepted 2 December 1998

Polymorphonuclear leukocytes (PMN) are essential for resolution of infections with Listeria monocytogenes. The present study investigated the role of the listerial exotoxins listeriolysin (LLO) and phosphatidylinositol-specific phospholipase C (PlcA) in human neutrophil activation. Different Listeria strains, mutated in individual virulence genes, as well as purified LLO were used. Coincubation of human neutrophils with wild-type L. monocytogenes provoked PMN activation, occurring independently of phagocytosis events, with concomitant elastase secretion, leukotriene generation, platelet-activating factor (PAF) synthesis, respiratory burst, and enhanced phosphoinositide hydrolysis. Degranulation and leukotriene formation were noted to be solely dependent on LLO expression, as these features were absent when the LLO-defective mutant EGD- and the avirulent strain L. innocua were used. These effects were fully reproduced by a recombinant L. innocua strain expressing LLO (INN+) and by the purified LLO molecule. LLO secretion was also required for PAF synthesis. However, wild-type L. monocytogenes was more potent in eliciting PAF formation than mutants expressing LLO, suggesting the involvement of additional virulence factors. This was even more obvious for phosphoinositide hydrolysis and respiratory burst: these events were provoked not only by INN+ but also by the LLO-defective mutant EGD- and by a recombinant L. innocua strain producing listerial PlcA. We conclude that human neutrophils react to extracellularly provided listerial exotoxins by rapid cell activation. Listeriolysin is centrally involved in triggering degranulation and lipid mediator generation, and further virulence factors such as PlcA apparently contribute to trigger neutrophil phosphoinositide hydrolysis and respiratory burst. In this way, listerial exotoxins may influence the host defense against infections with L. monocytogenes.

* Corresponding author. Mailing address: Department of Internal Medicine, Klinikstraße 36, D-35392 Giessen, Germany. Phone: 641-99-42351. Fax: 641-99-42359. E-mail: Friedrich.Grimminger{at}innere.med.uni-giessen.de.

Infection and Immunity, March 1999, p. 1125-1130, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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