The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1997/8/569/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 4, August 18, 1997 569-580

Article

Murine Salmonellosis Studied by Confocal Microscopy: Salmonella typhimurium Resides Intracellularly Inside Macrophages and Exerts a Cytotoxic Effect on Phagocytes In Vivo

Agneta Richter-Dahlfors*, Alison M.J. Buchan{ddagger}, and B. Brett Finlay*

From the * Biotechnology Laboratory and {ddagger} Department of Physiology, University of British Columbia, Vancouver, B.C., Canada V6T 1Z3

Salmonella typhimurium is considered a facultative intracellular pathogen, but its intracellular location in vivo has not been demonstrated conclusively. Here we describe the development of a new method to study the course of the histopathological processes associated with murine salmonellosis using confocal laser scanning microscopy of immunostained sections of mouse liver. Confocal microscopy of 30-µm-thick sections was used to detect bacteria after injection of ~100 CFU of S. typhimurium SL1344 intravenously into BALB/c mice, allowing salmonellosis to be studied in the murine model using more realistic small infectious doses. The appearance of bacteria in the mouse liver coincided in time and location with the infiltration of neutrophils in inflammatory foci. At later stages of disease the bacteria colocalized with macrophages and resided intracellularly inside these macrophages. Bacteria were cytotoxic for phagocytic cells, and apoptotic nuclei were detected immunofluorescently, whether phagocytes harbored intracellular bacteria or not. These data argue that Salmonella resides intracellularly inside macrophages in the liver and triggers cell death of phagocytes, processes which are involved in disease. This method is also applicable to other virulence models to examine infections at a cellular and subcellular level in vivo.

Address correspondence to Dr. B. Brett Finlay, Biotechnology Laboratory, University of British Columbia, Room 237-6174 University Boulevard, Vancouver, B.C., Canada V6T 1Z3. Phone: 604-822-2210; FAX: 604-822-9830; E-mail: bfinlay{at}unixg.ubc.ca

A. Richter-Dahlfors is supported by a postdoctoral fellowship from the Wenner-Gren Foundations, Stockholm, Sweden, and the work was supported by an operating grant to B.B. Finlay from the Medical Research Council of Canada. B.B. Finlay is an MRC Scientist and a Howard Hughes International Scholar.

1 Abbreviations used in this paper: CLSM, confocal laser scanning microscopy; TxR, Texas red.


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