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Research Paper |
Department of Infectious Diseases, Centre for Molecular Microbiology and Infection, Imperial College School of Medicine, Armstrong Road, London SW7 2AZ, UK1
Author for correspondence: David W. Holden. Tel: +44 20 7594 3073. Fax: +44 20 7594 3076. e-mail: d.holden{at}ic.ac.uk
Intracellular pathogens have developed different mechanisms which enable their survival and replication within the host cells. Some survive and replicate within a membrane-bound vacuole modified by the bacteria to support microbial growth (e.g. Salmonella enterica serovar Typhimurium), whereas others escape from the vacuole into the host cell cytosol, where they proliferate (e.g. Listeria monocytogenes). In this study a Salmonella strain carrying a mutation in sifA which is released from the vacuole was used to analyse Salmonella survival and replication within the cytosol of several cell lines. It was found that Salmonella replicates within the cytosol of epithelial cells at a higher rate than that achieved when replicating within the vacuole, but is defective for replication in the cytosol of fibroblasts or macrophages. Using an aroC purD double mutant strain which does not replicate within host cells, it was shown that Salmonella encounters a killing activity within the cytosol of macrophages. Furthermore, in vitro experiments using cytosol extracted from either infected or uninfected macrophages suggested that this activity is activated upon Salmonella infection.
Keywords: intracellular replication, type III secretion system, antimicrobial activity, host defence, intracellular pathogen
Abbreviations: FCS, fetal calf serum; GFP, green fluorescent protein; i.p., intraperitoneal(ly); PFA, paraformaldehyde; SCV, Salmonella-containing vacuole; SPI-2, Salmonella pathogenicity island 2; TRSC, Texas red sulfonyl chloride; TTSS, type III secretion system
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