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J Med Microbiol 55 (2006), 495-504; DOI: 10.1099/jmm.0.46390-0
© 2006 Society for General Microbiology
ISSN 1473-5644

Clostridium perfringens phospholipase C-induced platelet/leukocyte interactions impede neutrophil diapedesis

A. E. Bryant1,2, C. R. Bayer1,2, M. J. Aldape2, R. J. Wallace1, R. W. Titball3 and D. L. Stevens1,4

1 Infectious Diseases Section, Veterans Affairs Medical Center, Building 45, 500 West Fort Street, Boise, ID 83702, USA

2 University of Idaho, Moscow, ID, USA

3 Defence Science and Technology Laboratory, Porton Down, Salisbury, UK

4 University of Washington, Seattle, WA, USA

Correspondence
A. E. Bryant
amy.bryant{at}mindspring.com

Received 29 October 2005
Accepted 11 January 2006

Clostridium perfringens gas gangrene is a fulminant necrotizing infection in which inflammatory cells are notably absent from infected tissues but are often massed within adjacent vessels. It has been shown that C. perfringens phospholipase C (PLC) stimulates formation of large intravascular platelet/leukocyte complexes and that PLC-induced activation of platelet gpIIbIIIa plays a major role. In vivo, such aggregates contribute to microvascular thrombosis and ischaemic necrosis of tissue. However, the effects of adherent platelets on neutrophil diapedesis have not been established. The present work investigated (1) the contribution of platelet P-selectin (CD62P) to PLC-induced cellular complex formation and (2) the effects of platelet adhesion on neutrophil diapedesis. The effects of anti-gpIIbIIIa and anti-CD62P strategies on PLC-induced complex formation were measured by flow cytometry and followed by light microscopy. Both platelet gpIIbIIIa and CD62P contributed to the formation of platelet/leukocyte complexes. Specifically, gpIIbIIIa mediated the formation of large platelet/platelet aggregates that were tethered to the leukocyte principally via CD62P. Neutrophil diapedesis, quantified by a transendothelial cell migration assay and visualized by electron microscopy, was significantly reduced (>60 %) by the adherence of large platelet aggregates. It was concluded that the absence of a tissue inflammatory response in C. perfringens gas gangrene is due, in part, to impaired neutrophil mobility caused by large aggregates of adherent platelets induced by PLC. Further, an adjunctive immunotherapeutic strategy targeting both gpIIbIIIa and CD62P may improve the tissue inflammatory response, prevent vascular occlusion, maintain tissue viability, and reduce the need for radical amputation in patients with clostridial gas gangrene.

Abbreviations: MFI, mean fluorescence intensity; PE, phycoerythrin; PLC, phospholipase C; PMNL, polymorphonuclear leukocytes.




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