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Infection and Immunity, January 1999, p. 213-219, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Influence of the Salmonella typhimurium
Pathogenicity Island 2 Type III Secretion System on Bacterial Growth in
the Mouse
Jacqueline E.
Shea,
Carmen R.
Beuzon,
Colin
Gleeson,
Rosanna
Mundy, and
David W.
Holden*
Department of Infectious Diseases, Imperial
College School of Medicine, Hammersmith Hospital, London W12 0NN,
United Kingdom
Received 8 September 1998/Returned for modification 6 October
1998/Accepted 20 October 1998
We have investigated the in vivo growth kinetics of a
Salmonella typhimurium strain (P11D10) carrying a mutation
in ssaJ, a Salmonella pathogenicity island 2 (SPI2) gene encoding a component of a type III secretion system
required for systemic growth in mice. Similar numbers of mutant and
wild-type cells were recovered from the spleens and livers of BALB/c
mice up to 8 h after inoculation by the intraperitoneal route.
Thereafter, the numbers of wild-type cells continued to increase
logarithmically in these organs, whereas those of P11D10 remained
relatively static for several days before being cleared. Gentamicin
protection experiments on spleen cell suspensions recovered from
infected mice showed that viable intracellular wild-type bacteria
accumulated over time but that intracellular P11D10 cells did not.
Infection experiments were also performed with wild-type and P11D10
cells carrying the temperature-sensitive plasmid pHSG422 to distinguish
between bacterial growth rates and killing in vivo. At 16 h
postinoculation there were 10-fold more wild-type cells than mutant
cells in the spleens of infected mice, but the numbers of cells of both
strains carrying the nonreplicating plasmid were very similar, showing
that there was little difference in the degree of killing sustained by
the two strains and that the SPI2 secretion system must be required for
bacterial replication, rather than survival, in vivo. The SPI2 mutant
phenotype in mice is similar to that of strains carrying mutations in
the Salmonella virulence plasmid spv genes. To
determine if these two sets of genes interact together, a double mutant
strain carrying SPI2 and spv mutations was constructed and
compared with strains carrying single mutations in terms of virulence
attenuation. These experiments failed to provide any evidence showing
that the SPI2 and spv gene products interact together as
part of the same virulence mechanism.
*
Corresponding author. Mailing address: Department of
Infectious Diseases, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Rd., London W12 0NN, United Kingdom. Phone: 44-181-383-3487. Fax: 44-181-383-2078. E-mail:
dholden{at}rpms.ac.uk.
Infection and Immunity, January 1999, p. 213-219, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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