The characterization of the immune response of chickens to Salmonella infection is usually limited to the quantification of expression of genes coding for cytokines, chemokines or antimicrobial peptides. However, processes occurring in the cecum of infected chickens are likely to be much more diverse. In this study we have therefore characterized the transcriptome and proteome in the chicken cecum after infection with Salmonella Enteritidis. Using a combination of 454 pyrosequencing, protein mass spectrometry and quantitative real-time PCR, we identified 48 down- and 56 up-regulated chicken genes after Salmonella Enteritidis infection. The most inducible gene was that coding for MMP7, exhibiting a 5952 fold induction 9 days post-infection. An induction of greater than 100 fold was observed for IgG, IRG1, SAA, ExFABP, IL-22, TRAP6, MRP126, IFNγ, iNOS, ES1, IL-1β, LYG2, IFIT5, IL-17, AVD, AH221 and SERPIN B. Since prostaglandin D2 synthase was upregulated and degrading hydroxyprostaglandin dehydrogenase was downregulated after the infection, prostaglandin must accumulate in the cecum of chickens infected with Salmonella Enteritidis. Finally, above mentioned signaling was dependent on the presence of a SPI1-encoded type III secretion system in Salmonella Enteritidis. The inflammation lasted for 2 weeks after which time the expression of the "inflammatory" genes returned back to basal levels and, instead, the expression of IgA and IgG increased. This points to an important role for immunoglobulins in the restoration of homeostasis in the cecum after infection.

Veterinary Research Institute Hudcova 70 Brno 621 00 Czech Republic

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Anonymous. Report of the task force on zoonoses data collection on the analysis of the baseline study on the prevalence of Salmonella in holdings of laying hen flocks of Gallus gallus. EFSA J. 2007;97:1–84.

Lahuerta A, Westrell T, Takkinen J, Boelaert F, Rizzi V, Helwigh B, Borck B, Korsgaard H, Ammon A, Makela P. Zoonoses in the European Union: origin, distribution and dynamics - the EFSA-ECDC summary report 2009. Euro Surveill. 2011;16(13) Available online: http://www.eurosurveillance.org/ViewArticle.aspx?ArticleId=19832. PubMed

Beal RK, Powers C, Wigley P, Barrow PA, Smith AL. Temporal dynamics of the cellular, humoral and cytokine responses in chickens during primary and secondary infection with Salmonella enterica serovar Typhimurium. Avian Pathol. 2004;33:25–33. doi: 10.1080/03079450310001636282. PubMed DOI

Berndt A, Wilhelm A, Jugert C, Pieper J, Sachse K, Methner U. Chicken cecum immune response to Salmonella enterica serovars of different levels of invasiveness. Infect Immun. 2007;75:5993–6007. doi: 10.1128/IAI.00695-07. PubMed DOI PMC

Crhanova M, Hradecka H, Faldynova M, Matulova M, Havlickova H, Sisak F, Rychlik I. Immune response of chicken gut to natural colonization by gut microflora and to Salmonella enterica serovar enteritidis infection. Infect Immun. 2011;79:2755–2763. doi: 10.1128/IAI.01375-10. PubMed DOI PMC

Matulova M, Rajova J, Vlasatikova L, Volf J, Stepanova H, Havlickova H, Sisak F, Rychlik I. Characterization of chicken spleen transcriptome after infection with Salmonella enterica serovar Enteritidis. PLoS One. 2012;7:e48101. doi: 10.1371/journal.pone.0048101. PubMed DOI PMC

Raffatellu M, George MD, Akiyama Y, Hornsby MJ, Nuccio SP, Paixao TA, Butler BP, Chu H, Santos RL, Berger T, Mak TW, Tsolis RM, Bevins CL, Solnick JV, Dandekar S, Bäumler AJ. Lipocalin-2 resistance confers an advantage to Salmonella enterica serotype Typhimurium for growth and survival in the inflamed intestine. Cell Host Microbe. 2009;5:476–486. doi: 10.1016/j.chom.2009.03.011. PubMed DOI PMC

Matulova M, Havlickova H, Sisak F, Rychlik I. Vaccination of chickens with Salmonella Pathogenicity Island (SPI) 1 and SPI2 defective mutants of Salmonella enterica serovar Enteritidis. Vaccine. 2012;30:2090–2097. doi: 10.1016/j.vaccine.2012.01.050. PubMed DOI

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

Wisniewski JR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nat Methods. 2009;6:359–362. doi: 10.1038/nmeth.1322. PubMed DOI

Wisniewski JR, Nagaraj N, Zougman A, Gnad F, Mann M. Brain phosphoproteome obtained by a FASP-based method reveals plasma membrane protein topology. J Proteome Res. 2010;9:3280–3289. doi: 10.1021/pr1002214. PubMed DOI

Hsu JL, Huang SY, Chen SH. Dimethyl multiplexed labeling combined with microcolumn separation and MS analysis for time course study in proteomics. Electrophoresis. 2006;27:3652–3660. doi: 10.1002/elps.200600147. PubMed DOI

Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000;132:365–386. PubMed

Pilousova L, Rychlik I. Retron Se72 utilizes a unique strategy of the self-priming initiation of reverse transcription. Cell Mol Life Sci. 2011;68:3607–3617. doi: 10.1007/s00018-011-0671-0. PubMed DOI PMC

Stanford School of Medicine, Ashley Lab in the Department of Medicine. [ http://ashleylab.stanford.edu/tools_scripts.html]

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

Matulova M, Stepanova H, Sisak F, Havlickova H, Faldynova M, Kyrova K, Volf J, Rychlik I. Cytokine signaling in splenic leukocytes from vaccinated and non-vaccinated chickens after intravenous infection with Salmonella Enteritidis. PLoS One. 2012;7:e32346. doi: 10.1371/journal.pone.0032346. PubMed DOI PMC

McPherson M, Wei B, Turovskaya O, Fujiwara D, Brewer S, Braun J. Colitis immunoregulation by CD8+ T cell requires T cell cytotoxicity and B cell peptide antigen presentation. Am J Physiol Gastrointest Liver Physiol. 2008;295:G485–G492. doi: 10.1152/ajpgi.90221.2008. PubMed DOI PMC

Wei B, McPherson M, Turovskaya O, Velazquez P, Fujiwara D, Brewer S, Braun J. Integration of B cells and CD8+ T in the protective regulation of systemic epithelial inflammation. Clin Immunol. 2008;127:303–312. doi: 10.1016/j.clim.2008.01.001. PubMed DOI PMC

Chowdhury SR, King DE, Willing BP, Band MR, Beever JE, Lane AB, Loor JJ, Marini JC, Rund LA, Schook LB, Van Kessel AG, Gaskins HR. Transcriptome profiling of the small intestinal epithelium in germfree versus conventional piglets. BMC Genomics. 2007;8:215. doi: 10.1186/1471-2164-8-215. PubMed DOI PMC

Zhao SH, Kuhar D, Lunney JK, Dawson H, Guidry C, Uthe JJ, Bearson SM, Recknor J, Nettleton D, Tuggle CK. Gene expression profiling in Salmonella Choleraesuis-infected porcine lung using a long oligonucleotide microarray. Mamm Genome. 2006;17:777–789. doi: 10.1007/s00335-005-0155-3. PubMed DOI

Kitamura D, Kaneko H, Miyagoe Y, Ariyasu T, Watanabe T. Isolation and characterization of a novel human gene expressed specifically in the cells of hematopoietic lineage. Nucleic Acids Res. 1989;17:9367–9379. PubMed PMC

Yamanashi Y, Okada M, Semba T, Yamori T, Umemori H, Tsunasawa S, Toyoshima K, Kitamura D, Watanabe T, Yamamoto T. Identification of HS1 protein as a major substrate of protein-tyrosine kinase(s) upon B-cell antigen receptor-mediated signaling. Proc Natl Acad Sci U S A. 1993;90:3631–3635. doi: 10.1073/pnas.90.8.3631. PubMed DOI PMC

Hao JJ, Carey GB, Zhan X. Syk-mediated tyrosine phosphorylation is required for the association of hematopoietic lineage cell-specific protein 1 with lipid rafts and B cell antigen receptor signalosome complex. J Biol Chem. 2004;279:33413–33420. doi: 10.1074/jbc.M313564200. PubMed DOI

Taniuchi I, Kitamura D, Maekawa Y, Fukuda T, Kishi H, Watanabe T. Antigen-receptor induced clonal expansion and deletion of lymphocytes are impaired in mice lacking HS1 protein, a substrate of the antigen-receptor-coupled tyrosine kinases. EMBO J. 1995;14:3664–3678. PubMed PMC

Defendenti C, Sarzi-Puttini P, Grosso S, Croce A, Senesi O, Saibeni S, Bollani S, Almasio PL, Bruno S, Atzeni F. B lymphocyte intestinal homing in inflammatory bowel disease. BMC Immunol. 2011;12:71. doi: 10.1186/1471-2172-12-71. PubMed DOI PMC

Suzuki K, Meek B, Doi Y, Muramatsu M, Chiba T, Honjo T, Fagarasan S. Aberrant expansion of segmented filamentous bacteria in IgA-deficient gut. Proc Natl Acad Sci U S A. 2004;101:1981–1986. doi: 10.1073/pnas.0307317101. PubMed DOI PMC

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