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Infection and Immunity, November 1999, p. 5690-5698, Vol. 67, No. 11
Departments of Microbiology and Immunology
and the Biotechnology Laboratory, Vancouver, British Columbia,
Canada V6T 1Z3
Received 12 April 1999/Returned for modification 23 June
1999/Accepted 25 August 1999
Survival and growth of salmonellae within host cells are important
aspects of bacterial virulence. We have developed an assay to identify
Salmonella typhimurium genes that are induced inside Salmonella-containing vacuoles within macrophage and
epithelial cells. A promoterless luciferase gene cassette was inserted
randomly into the Salmonella chromosome, and the resulting
mutants were screened for genes upregulated in intracellular bacteria
compared to extracellular bacteria. We identified four genes in
S. typhimurium that were upregulated upon bacterial
invasion of both phagocytic and nonphagocytic cells. Expression of
these genes was not induced by factors secreted by host cells or media
alone. All four genes were induced at early time points (2 to 4 h)
postinvasion and continued to be upregulated within host cells at later
times (5 to 7 h). One mutant contained an insertion in the
ssaR gene, within Salmonella pathogenicity
island 2 (SPI-2), which abolished bacterial virulence in a murine
typhoid model. Two other mutants contained insertions within SPI-5, one
in the sopB/sigD gene and the other in a downstream gene,
pipB. The insertions within SPI-5 resulted in the
attenuation of S. typhimurium in the mouse model. The
fourth mutant contained an insertion within a previously undescribed region of the S. typhimurium chromosome, iicA
(induced intracellularly A). We detected no effect on virulence as a
result of this insertion. In conclusion, all but one of the genes
identified in this study were virulence factors within pathogenicity
islands, illustrating the requirement for specific gene expression
inside mammalian cells and indicating the key role that virulence
factor regulation plays in Salmonella pathogenesis.
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Salmonella typhimurium Virulence Genes Are Induced
upon Bacterial Invasion into Phagocytic and Nonphagocytic
Cells
*
Corresponding author. Mailing address: Biotechnology
Laboratory, Room 237, 6174 University Blvd., Vancouver, BC, Canada V6T 1Z3. Phone: (604) 822-2210. Fax: (604) 822-9830. E-mail:
bfinlay{at}unixg.ubc.ca.
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