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. 1993 Dec;61(12):5231–5236. doi: 10.1128/iai.61.12.5231-5236.1993

Expression of the Salmonella virulence plasmid gene spvB in cultured macrophages and nonphagocytic cells.

J Fierer 1, L Eckmann 1, F Fang 1, C Pfeifer 1, B B Finlay 1, D Guiney 1
PMCID: PMC281306  PMID: 8225598

Abstract

Certain serotypes of salmonellae carry virulence plasmids that greatly enhance the pathogenicity of these bacteria in experimentally infected mice. This phenotype is largely attributable to the 8-kb spv regulon. However, spv genes are not expressed while bacteria grow in vitro. We now show that spvB, which is required for virulence, is expressed rapidly after Salmonella dublin is ingested by cultured J774 and murine peritoneal macrophages and that expression is not affected by the alkalinization of intracellular vesicles. The level of induction of spvB is reduced when macrophages are pretreated with gamma interferon. spvB is also expressed in human and canine epithelial cell lines and a human hepatoma cell line. In all cases, spvB expression is dependent on the spvR gene, just as it is in stationary-phase cultures in vitro. These data suggest that spv virulence genes are expressed by intracellular salmonellae in vivo in response to a signal that is common to the intracellular compartments of cells that are invaded by salmonellae.

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Selected References

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  1. Adams D. O., Hamilton T. A. The cell biology of macrophage activation. Annu Rev Immunol. 1984;2:283–318. doi: 10.1146/annurev.iy.02.040184.001435. [DOI] [PubMed] [Google Scholar]
  2. Alpuche Aranda C. M., Swanson J. A., Loomis W. P., Miller S. I. Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10079–10083. doi: 10.1073/pnas.89.21.10079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barrow P. A., Lovell M. A. Functional homology of virulence plasmids in Salmonella gallinarum, S. pullorum, and S. typhimurium. Infect Immun. 1989 Oct;57(10):3136–3141. doi: 10.1128/iai.57.10.3136-3141.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beninger P. R., Chikami G., Tanabe K., Roudier C., Fierer J., Guiney D. G. Physical and genetic mapping of the Salmonella dublin virulence plasmid pSDL2. Relationship to plasmids from other Salmonella strains. J Clin Invest. 1988 May;81(5):1341–1347. doi: 10.1172/JCI113461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buchmeier N. A., Heffron F. Induction of Salmonella stress proteins upon infection of macrophages. Science. 1990 May 11;248(4956):730–732. doi: 10.1126/science.1970672. [DOI] [PubMed] [Google Scholar]
  6. Chikami G. K., Fierer J., Guiney D. G. Plasmid-mediated virulence in Salmonella dublin demonstrated by use of a Tn5-oriT construct. Infect Immun. 1985 Nov;50(2):420–424. doi: 10.1128/iai.50.2.420-424.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conlan J. W., North R. J. Early pathogenesis of infection in the liver with the facultative intracellular bacteria Listeria monocytogenes, Francisella tularensis, and Salmonella typhimurium involves lysis of infected hepatocytes by leukocytes. Infect Immun. 1992 Dec;60(12):5164–5171. doi: 10.1128/iai.60.12.5164-5171.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fang F. C., Krause M., Roudier C., Fierer J., Guiney D. G. Growth regulation of a Salmonella plasmid gene essential for virulence. J Bacteriol. 1991 Nov;173(21):6783–6789. doi: 10.1128/jb.173.21.6783-6789.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fang F. C., Libby S. J., Buchmeier N. A., Loewen P. C., Switala J., Harwood J., Guiney D. G. The alternative sigma factor katF (rpoS) regulates Salmonella virulence. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11978–11982. doi: 10.1073/pnas.89.24.11978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fields P. I., Swanson R. V., Haidaris C. G., Heffron F. Mutants of Salmonella typhimurium that cannot survive within the macrophage are avirulent. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5189–5193. doi: 10.1073/pnas.83.14.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fierer J., Hatlen L., Lin J. P., Estrella D., Mihalko P., Yau-Young A. Successful treatment using gentamicin liposomes of Salmonella dublin infections in mice. Antimicrob Agents Chemother. 1990 Feb;34(2):343–348. doi: 10.1128/aac.34.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fierer J., Krause M., Tauxe R., Guiney D. Salmonella typhimurium bacteremia: association with the virulence plasmid. J Infect Dis. 1992 Sep;166(3):639–642. doi: 10.1093/infdis/166.3.639. [DOI] [PubMed] [Google Scholar]
  13. Finlay B. B., Falkow S. Salmonella as an intracellular parasite. Mol Microbiol. 1989 Dec;3(12):1833–1841. doi: 10.1111/j.1365-2958.1989.tb00170.x. [DOI] [PubMed] [Google Scholar]
  14. Friedlander A. M. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem. 1986 Jun 5;261(16):7123–7126. [PubMed] [Google Scholar]
  15. Garcia-del Portillo F., Foster J. W., Maguire M. E., Finlay B. B. Characterization of the micro-environment of Salmonella typhimurium-containing vacuoles within MDCK epithelial cells. Mol Microbiol. 1992 Nov;6(22):3289–3297. doi: 10.1111/j.1365-2958.1992.tb02197.x. [DOI] [PubMed] [Google Scholar]
  16. Groisman E. A., Chiao E., Lipps C. J., Heffron F. Salmonella typhimurium phoP virulence gene is a transcriptional regulator. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7077–7081. doi: 10.1073/pnas.86.18.7077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gulig P. A., Caldwell A. L., Chiodo V. A. Identification, genetic analysis and DNA sequence of a 7.8-kb virulence region of the Salmonella typhimurium virulence plasmid. Mol Microbiol. 1992 May;6(10):1395–1411. doi: 10.1111/j.1365-2958.1992.tb00860.x. [DOI] [PubMed] [Google Scholar]
  18. Gulig P. A., Curtiss R., 3rd Plasmid-associated virulence of Salmonella typhimurium. Infect Immun. 1987 Dec;55(12):2891–2901. doi: 10.1128/iai.55.12.2891-2901.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gulig P. A., Doyle T. J. The Salmonella typhimurium virulence plasmid increases the growth rate of salmonellae in mice. Infect Immun. 1993 Feb;61(2):504–511. doi: 10.1128/iai.61.2.504-511.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gulig P. A. Virulence plasmids of Salmonella typhimurium and other salmonellae. Microb Pathog. 1990 Jan;8(1):3–11. doi: 10.1016/0882-4010(90)90003-9. [DOI] [PubMed] [Google Scholar]
  21. Heffernan E. J., Fierer J., Chikami G., Guiney D. Natural history of oral Salmonella dublin infection in BALB/c mice: effect of an 80-kilobase-pair plasmid on virulence. J Infect Dis. 1987 Jun;155(6):1254–1259. doi: 10.1093/infdis/155.6.1254. [DOI] [PubMed] [Google Scholar]
  22. Jones G. W., Rabert D. K., Svinarich D. M., Whitfield H. J. Association of adhesive, invasive, and virulent phenotypes of Salmonella typhimurium with autonomous 60-megadalton plasmids. Infect Immun. 1982 Nov;38(2):476–486. doi: 10.1128/iai.38.2.476-486.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Korpela K., Ranki M., Sukupolvi S., Mäkelä P. H., Rhen M. Occurrence of Salmonella typhimurium virulence plasmid-specific sequences in different serovars of Salmonella. FEMS Microbiol Lett. 1989 Mar;49(1):49–54. doi: 10.1016/0378-1097(89)90340-6. [DOI] [PubMed] [Google Scholar]
  24. Krause M., Fang F. C., Guiney D. G. Regulation of plasmid virulence gene expression in Salmonella dublin involves an unusual operon structure. J Bacteriol. 1992 Jul;174(13):4482–4489. doi: 10.1128/jb.174.13.4482-4489.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Krause M., Roudier C., Fierer J., Harwood J., Guiney D. Molecular analysis of the virulence locus of the Salmonella dublin plasmid pSDL2. Mol Microbiol. 1991 Feb;5(2):307–316. doi: 10.1111/j.1365-2958.1991.tb02111.x. [DOI] [PubMed] [Google Scholar]
  26. Lange R., Hengge-Aronis R. Identification of a central regulator of stationary-phase gene expression in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):49–59. doi: 10.1111/j.1365-2958.1991.tb01825.x. [DOI] [PubMed] [Google Scholar]
  27. Lomholt H., van Alphen L., Kilian M. Antigenic variation of immunoglobulin A1 proteases among sequential isolates of Haemophilus influenzae from healthy children and patients with chronic obstructive pulmonary disease. Infect Immun. 1993 Nov;61(11):4575–4581. doi: 10.1128/iai.61.11.4575-4581.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mahan M. J., Slauch J. M., Mekalanos J. J. Selection of bacterial virulence genes that are specifically induced in host tissues. Science. 1993 Jan 29;259(5095):686–688. doi: 10.1126/science.8430319. [DOI] [PubMed] [Google Scholar]
  29. McFarland W. C., Stocker B. A. Effect of different purine auxotrophic mutations on mouse-virulence of a Vi-positive strain of Salmonella dublin and of two strains of Salmonella typhimurium. Microb Pathog. 1987 Aug;3(2):129–141. doi: 10.1016/0882-4010(87)90071-4. [DOI] [PubMed] [Google Scholar]
  30. Miller S. I., Kukral A. M., Mekalanos J. J. A two-component regulatory system (phoP phoQ) controls Salmonella typhimurium virulence. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5054–5058. doi: 10.1073/pnas.86.13.5054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Montenegro M. A., Morelli G., Helmuth R. Heteroduplex analysis of Salmonella virulence plasmids and their prevalence in isolates of defined sources. Microb Pathog. 1991 Dec;11(6):391–397. doi: 10.1016/0882-4010(91)90035-9. [DOI] [PubMed] [Google Scholar]
  32. Murray H. W. Interferon-gamma, the activated macrophage, and host defense against microbial challenge. Ann Intern Med. 1988 Apr;108(4):595–608. doi: 10.7326/0003-4819-108-4-595. [DOI] [PubMed] [Google Scholar]
  33. Pardon P., Popoff M. Y., Coynault C., Marly J., Miras I. Virulence-associated plasmids of Salmonella serotype Typhimurium in experimental murine infection. Ann Inst Pasteur Microbiol. 1986 Jul-Aug;137B(1):47–60. doi: 10.1016/s0769-2609(86)80093-x. [DOI] [PubMed] [Google Scholar]
  34. Popoff M. Y., Miras I., Coynault C., Lasselin C., Pardon P. Molecular relationships between virulence plasmids of Salmonella serotypes typhimurium and dublin and large plasmids of other Salmonella serotypes. Ann Microbiol (Paris) 1984 May-Jun;135A(3):389–398. doi: 10.1016/s0769-2609(84)80080-0. [DOI] [PubMed] [Google Scholar]
  35. Potter M., O'Brien A. D., Skamene E., Gros P., Forget A., Kongshavn P. A., Wax J. S. A BALB/c congenic strain of mice that carries a genetic locus (Ityr) controlling resistance to intracellular parasites. Infect Immun. 1983 Jun;40(3):1234–1235. doi: 10.1128/iai.40.3.1234-1235.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Riikonen P., Mäkelä P. H., Saarilahti H., Sukupolvi S., Taira S., Rhen M. The virulence plasmid does not contribute to growth of Salmonella in cultured murine macrophages. Microb Pathog. 1992 Oct;13(4):281–291. doi: 10.1016/0882-4010(92)90038-p. [DOI] [PubMed] [Google Scholar]
  37. Roudier C., Fierer J., Guiney D. G. Characterization of translation termination mutations in the spv operon of the Salmonella virulence plasmid pSDL2. J Bacteriol. 1992 Oct;174(20):6418–6423. doi: 10.1128/jb.174.20.6418-6423.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Roudier C., Krause M., Fierer J., Guiney D. G. Correlation between the presence of sequences homologous to the vir region of Salmonella dublin plasmid pSDL2 and the virulence of twenty-two Salmonella serotypes in mice. Infect Immun. 1990 May;58(5):1180–1185. doi: 10.1128/iai.58.5.1180-1185.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Valone S. E., Chikami G. K., Miller V. L. Stress induction of the virulence proteins (SpvA, -B, and -C) from native plasmid pSDL2 of Salmonella dublin. Infect Immun. 1993 Feb;61(2):705–713. doi: 10.1128/iai.61.2.705-713.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Williamson C. M., Baird G. D., Manning E. J. A common virulence region on plasmids from eleven serotypes of Salmonella. J Gen Microbiol. 1988 Apr;134(4):975–982. doi: 10.1099/00221287-134-4-975. [DOI] [PubMed] [Google Scholar]
  41. Williamson C. M., Pullinger G. D., Lax A. J. Identification of an essential virulence region on Salmonella plasmids. Microb Pathog. 1988 Dec;5(6):469–473. doi: 10.1016/0882-4010(88)90008-3. [DOI] [PubMed] [Google Scholar]
  42. van Dissel J. T., Stikkelbroeck J. J., Michel B. C., van den Barselaar M. T., Leijh P. C., van Furth R. Inability of recombinant interferon-gamma to activate the antibacterial activity of mouse peritoneal macrophages against Listeria monocytogenes and Salmonella typhimurium. J Immunol. 1987 Sep 1;139(5):1673–1678. [PubMed] [Google Scholar]

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