BACKGROUND: In this study we were interested in the colonisation and early immune response of Balb/C mice to infection with Salmonella Enteritidis and isogenic pathogenicity island free mutants. RESULTS: The virulence of S. Enteritidis for Balb/C mice was exclusively dependent on intact SPI-2. Infections with any of the mutants harbouring SPI-2 (including the mutant in which we left only SPI-2 but removed SPI-1, SPI-3, SPI-4 and SPI-5) resulted in fatalities, liver injures and NK cell depletion from the spleen. The infection was of minimal influence on counts of splenic CD4 CD8 T lymphocytes and gammadelta T-lymphocytes although a reduced ability of splenic lymphocytes to respond to non-specific mitogens indicated general immunosuppression in mice infected with SPI-2 positive S. Enteritidis mutants. Further investigations showed that NK cells were depleted also in blood but not in the caecal lamina propria. However, NK cell depletion was not directly associated with the presence of SPI-2 and was rather an indicator of virulence or avirulence of a particular mutant because the depletion was not observed in mice infected with other attenuated mutants such as lon and rfaL. CONCLUSIONS: The virulence of S. Enteritidis for Balb/C mice is exclusively dependent on the presence of SPI-2 in its genome, and a major hallmark of the infection in terms of early changes in lymphocyte populations is the depletion of NK cells in spleen and blood. The decrease of NK cells in circulation can be used as a marker of attenuation of S. Enteritidis mutants for Balb/C mice.

Veterinary Research Institute Hudcova 70 62100 Brno Czech Republic

Zobrazit více v PubMed

Mills DM, Bajaj V, Lee CA. A 40 kb chromosomal fragment encoding Salmonella typhimurium invasion genes is absent from the corresponding region of the Escherichia coli K-12 chromosome. Mol Microbiol. 1995;15:749–759. doi: 10.1111/j.1365-2958.1995.tb02382.x. PubMed DOI

Bajaj V, Lucas RL, Hwang C, Lee CA. Co-ordinate regulation of Salmonella typhimurium invasion genes by environmental and regulatory factors is mediated by control of hilA expression. Mol Microbiol. 1996;22:703–714. doi: 10.1046/j.1365-2958.1996.d01-1718.x. PubMed DOI

Cirillo DM, Valdivia RH, Monack DM, Falkow S. Macrophage-dependent induction of the Salmonella pathogenicity island 2 type III secretion system and its role in intracellular survival. Mol Microbiol. 1998;30:175–188. doi: 10.1046/j.1365-2958.1998.01048.x. PubMed DOI

Blanc-Potard AB, Groisman EA. The Salmonella selC locus contains a pathogenicity island mediating intramacrophage survival. EMBO J. 1997;16:5376–5385. doi: 10.1093/emboj/16.17.5376. PubMed DOI PMC

Morgan E, Campbell JD, Rowe SC, Bispham J, Stevens MP, Bowen AJ, Barrow PA, Maskell DJ, Wallis TS. Identification of host-specific colonization factors of Salmonella enterica serovar Typhimurium. Mol Microbiol. 2004;54:994–1010. doi: 10.1111/j.1365-2958.2004.04323.x. PubMed DOI

Kiss T, Morgan E, Nagy G. Contribution of SPI-4 genes to the virulence of Salmonella enterica. FEMS Microbiol Lett. 2007;275:153–159. doi: 10.1111/j.1574-6968.2007.00871.x. PubMed DOI

Knodler LA, Celli J, Hardt WD, Vallance BA, Yip C, Finlay BB. Salmonella effectors within a single pathogenicity island are differentially expressed and translocated by separate type III secretion systems. Mol Microbiol. 2002;43:1089–1103. doi: 10.1046/j.1365-2958.2002.02820.x. PubMed DOI

Papezova K, Gregorova D, Jonuschies J, Rychlik I. Ordered expression of virulence genes in Salmonella enterica serovar typhimurium. Folia Microbiol (Praha) 2007;52:107–114. doi: 10.1007/BF02932148. PubMed DOI

Kaiser P, Rothwell L, Galyov EE, Barrow PA, Burnside J, Wigley P. Differential cytokine expression in avian cells in response to invasion by Salmonella typhimurium, Salmonella enteritidis and Salmonella gallinarum. Microbiology. 2000;146(Pt 12):3217–3226. PubMed

Zhang S, Adams LG, Nunes J, Khare S, Tsolis RM, Baumler AJ. Secreted effector proteins of Salmonella enterica serotype typhimurium elicit host-specific chemokine profiles in animal models of typhoid fever and enterocolitis. Infect Immun. 2003;71:4795–4803. doi: 10.1128/IAI.71.8.4795-4803.2003. PubMed DOI PMC

Wigley P, Hulme S, Powers C, Beal R, Smith A, Barrow P. Oral infection with the Salmonella enterica serovar Gallinarum 9R attenuated live vaccine as a model to characterise immunity to fowl typhoid in the chicken. BMC Vet Res. 2005;1:2. doi: 10.1186/1746-6148-1-2. PubMed DOI PMC

Geddes K, Cruz F, Heffron F. Analysis of cells targeted by Salmonella type III secretion in vivo. PLoS Pathog. 2007;3:e196. doi: 10.1371/journal.ppat.0030196. PubMed DOI PMC

Hersh D, Monack DM, Smith MR, Ghori N, Falkow S, Zychlinsky A. The Salmonella invasin SipB induces macrophage apoptosis by binding to caspase-1. Proc Natl Acad Sci USA. 1999;96:2396–2401. doi: 10.1073/pnas.96.5.2396. PubMed DOI PMC

Lundberg U, Vinatzer U, Berdnik D, von Gabain A, Baccarini M. Growth phase-regulated induction of Salmonella -induced macrophage apoptosis correlates with transient expression of SPI-1 genes. J Bacteriol. 1999;181:3433–3437. PubMed PMC

Halici S, Zenk SF, Jantsch J, Hensel M. Functional analysis of the Salmonella pathogenicity island 2-mediated inhibition of antigen presentation in dendritic cells. Infect Immun. 2008;76:4924–4933. doi: 10.1128/IAI.00531-08. PubMed DOI PMC

Kirby AC, Yrlid U, Wick MJ. The innate immune response differs in primary and secondary Salmonella infection. J Immunol. 2002;169:4450–4459. PubMed

Hensel M, Shea JE, Waterman SR, Mundy R, Nikolaus T, Banks G, Vazquez-Torres A, Gleeson C, Fang FC, Holden DW. Genes encoding putative effector proteins of the type III secretion system of Salmonella pathogenicity island 2 are required for bacterial virulence and proliferation in macrophages. Mol Microbiol. 1998;30:163–174. doi: 10.1046/j.1365-2958.1998.01047.x. PubMed DOI

Murray RA, Lee CA. Invasion genes are not required for Salmonella enterica serovar typhimurium to breach the intestinal epithelium: evidence that Salmonella pathogenicity island 1 has alternative functions during infection. Infect Immun. 2000;68:5050–5055. doi: 10.1128/IAI.68.9.5050-5055.2000. PubMed DOI PMC

Jiang X, Rossanese OW, Brown NF, Kujat-Choy S, Galan JE, Finlay BB, Brumell JH. The related effector proteins SopD and SopD2 from Salmonella enterica serovar Typhimurium contribute to virulence during systemic infection of mice. Mol Microbiol. 2004;54:1186–1198. doi: 10.1111/j.1365-2958.2004.04344.x. PubMed DOI

Pfeifer CG, Marcus SL, Steele-Mortimer O, Knodler LA, Finlay BB. Salmonella typhimurium virulence genes are induced upon bacterial invasion into phagocytic and nonphagocytic cells. Infect Immun. 1999;67:5690–5698. PubMed PMC

Kaniga K, Trollinger D, Galan JE. Identification of two targets of the type III protein secretion system encoded by the inv and spa loci of Salmonella typhimurium that have homology to the Shigella IpaD and IpaA proteins. J Bacteriol. 1995;177:7078–7085. PubMed PMC

Emoto M, Naito T, Nakamura R, Yoshikai Y. Different appearance of gamma delta T cells during salmonellosis between Ityr and Itys mice. J Immunol. 1993;150:3411–3420. PubMed

Grillon C, Monsigny M, Kieda C. Changes in the expression of lectins in human T lymphocyte membrane upon mitogenic stimulation. Carbohydr Res. 1991;213:283–292. doi: 10.1016/S0008-6215(00)90615-3. PubMed DOI

Harrington L, Srikanth CV, Antony R, Shi HN, Cherayil BJ. A role for natural killer cells in intestinal inflammation caused by infection with Salmonella enterica serovar Typhimurium. FEMS Immunol Med Microbiol. 2007;51:372–380. doi: 10.1111/j.1574-695X.2007.00313.x. PubMed DOI PMC

Lapaque N, Walzer T, Meresse S, Vivier E, Trowsdale J. Interactions between human NK cells and macrophages in response to Salmonella infection. J Immunol. 2009;182:4339–4348. doi: 10.4049/jimmunol.0803329. PubMed DOI

Perona-Wright G, Mohrs K, Szaba FM, Kummer LW, Madan R, Karp CL, Johnson LL, Smiley ST, Mohrs M. Systemic but not local infections elicit immunosuppressive IL-10 production by natural killer cells. Cell Host Microbe. 2009;6:503–512. doi: 10.1016/j.chom.2009.11.003. PubMed DOI PMC

Agaugue S, Marcenaro E, Ferranti B, Moretta L, Moretta A. Human natural killer cells exposed to IL-2, IL-12, IL-18, or IL-4 differently modulate priming of naive T cells by monocyte-derived dendritic cells. Blood. 2008;112:1776–1783. doi: 10.1182/blood-2008-02-135871. PubMed DOI

Methner U, Barrow PA, Gregorova D, Rychlik I. Intestinal colonisation-inhibition and virulence of Salmonella phoP, rpoS and ompC deletion mutants in chickens. Vet Microbiol. 2004;98:37–43. doi: 10.1016/j.vetmic.2003.10.019. PubMed DOI

Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA. 2000;97:6640–6645. doi: 10.1073/pnas.120163297. PubMed DOI PMC

Rychlik I, Karasova D, Sebkova A, Volf J, Sisak F, Havlickova H, Kummer V, Imre A, Szmolka A, Nagy B. Virulence potential of five major pathogenicity islands (SPI-1 to SPI-5) of Salmonella enterica serovar Enteritidis for chickens. BMC Microbiol. 2009;9:268. doi: 10.1186/1471-2180-9-268. PubMed DOI PMC

Methner U, al Shabibi S, Meyer H. Experimental oral infection of specific pathogen-free laying hens and cocks with Salmonella enteritidis strains. Zentralbl Veterinarmed B. 1995;42:459–469. PubMed

Faldyna M, Leva L, Knotigova P, Toman M. Lymphocyte subsets in peripheral blood of dogs--a flow cytometric study. Vet Immunol Immunopathol. 2001;82:23–37. doi: 10.1016/S0165-2427(01)00337-3. PubMed DOI

Karasova D, Sebkova A, Vrbas V, Havlickova H, Sisak F, Rychlik I. Comparative analysis of Salmonella enterica serovar Enteritidis mutants with a vaccine potential. Vaccine. 2009;27:5265–5270. doi: 10.1016/j.vaccine.2009.06.060. PubMed DOI

Overbergh L, Giulietti A, Valckx D, Decallonne R, Bouillon R, Mathieu C. The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression. J Biomol Tech. 2003;14:33–43. PubMed PMC

Immune protection of chickens conferred by a vaccine consisting of attenuated strains of Salmonella Enteritidis, Typhimurium and Infantis

Composition of Gut Microbiota Influences Resistance of Newly Hatched Chickens to Salmonella Enteritidis Infection

phoP, SPI1, SPI2 and aroA mutants of Salmonella Enteritidis induce a different immune response in chickens

Gene expression in the chicken caecum in response to infections with non-typhoid Salmonella

Vaccination of chickens with SPI1-lon and SPI1-lon-fliC mutant of Salmonella enterica Serovar Enteritidis

Cytokine signaling in splenic leukocytes from vaccinated and non-vaccinated chickens after intravenous infection with Salmonella enteritidis

SPI-1-encoded type III secretion system of Salmonella enterica is required for the suppression of porcine alveolar macrophage cytokine expression