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Infection and Immunity, December 2002, p. 7126-7135, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.7126-7135.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genetic Requirements for Salmonella-Induced Cytopathology in Human Monocyte-Derived Macrophages

Sara H. Browne, Marc L. Lesnick, and Donald G. Guiney*

Department of Medicine, School of Medicine, University of California at San Diego, La Jolla, California 92093-0640

Received 10 May 2002/ Returned for modification 9 July 2002/ Accepted 9 September 2002

Infection of human macrophages with Salmonella enterica serovar Typhimurium or Salmonella enterica serovar Dublin produces delayed cytotoxicity characterized by cell detachment and associated apoptosis. Using a site-specific mutant in the SpvB active site, we verify that the ADP-ribosylation activity of SpvB is required for delayed cytotoxicity in human macrophages infected with Salmonella. SipB and the type III protein secretion system (TTSS) encoded by Salmonella pathogenicity island 1 (SPI1) are not involved, whereas the SPI2 TTSS is absolutely required for SpvB-dependent cytotoxicity. Furthermore, we show that infection of macrophage cultures with wild-type or sipB mutant bacteria led to a complete loss of polymerized actin in over half of the cells after 24 h. In contrast, macrophages infected with the spvB or SPI2 (ssaV or ssaJ) mutant strain retained normal F-actin filaments, despite similar numbers of intracellular bacteria. We conclude that SpvB and a functional SPI2 TTSS are essential for Salmonella-induced delayed cytotoxicity of human macrophages.

* Corresponding author. Mailing address: 9500 Gilman Dr., MC 0640, University of California at San Diego, La Jolla, CA 92093-0640. Phone: (858) 534-6030. Fax: (858) 534-6020. E-mail: dguiney{at}ucsd.edu.

Editor: D. L. Burns

Infection and Immunity, December 2002, p. 7126-7135, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.7126-7135.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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