Food Safety Research Information Office
Title:A Systems Biology Approach to Understanding the Salmonella-Host Interactome in Poultry and Swine
Objective:

Objective 1: Conduct research on the differential host-pathogen interactions of Salmonella in human, chicken, and swine intestinal key host immune cells using emerging genomic technologies.

Objective 2: Analyze and characterize both host and Salmonella proteins that are modulated in expression during infection using quantitative proteomics.

Objective 3: Research on the molecular and cellular details of the host-microbe interactions will be used to identify virulence-associated microbial genes and host defense strategies. Identify potential intervention targets (e.g., host kinases) for Salmonella infections in food animals.

Objective 4: Develop strategies for the reduction of foodborne pathogens by targeting the host innate immune system (by identifying the use of immunomodulatory antimicrobial or host-defense peptides) and targeting and identifying virulence factors. Sub-objective 4A: Molecular characterization of anti-infectives that target the host innate immune system to facilitate pathogen-specific immune responses. Sub-objective 4B: Develop a high-throughput assay to screen a series of commercial libraries of small molecules for their ability to inhibit virulence factors produced by S. typhimurium.

More Info:
Approach:

Objective 1: Utilize deep sequencing and mutagenesis technologies to dissect the differential host-pathogen interactions of Salmonella in human, chicken, and swine intestinal epithelial cells and macrophages. Specifically, we will determine the differential transcriptome of S. Typhimurium in mammalian versus chicken epithelial cells and develop gene-deletion mutants of S. Typhimurium to elucidate the differential mechanisms of intestinal pathogenicity of S. Typhimurium in humans compared to chickens and swine.

Objective 2: A newly described technique of purifying live Salmonella expressing green fluorescent protein from either infected tissues or cell cultures using flow cytometry will be used to analyze and characterize both host and Salmonella proteins that are modulated in expression during infection. Quantitative proteomics including conventional two-dimensional electrophoresis, difference gel electrophoresis (DIGE), and mass spectrometry will be used to facilitate accomplishment of this objective.

Objective 3: Using a matched comparative model (Salmonella characterized by contrasting degrees of pathogenicity and/or gene-deletion mutants of S. Typhimurium), newly developed peptide arrays will be used for studying the kinome of chicken and swine intestinal epithelial cells and macrophages. Cell lysates will be analyzed on a kinomics array containing 1,024 peptides derived from known phosphorylation sites annotated with reported upstream kinases. In addition, reverse chemical genetics will be used to identify host kinases that are essential in controlling intracellular Salmonella infections. This procedure will enable us to identify a class of kinases using selective chemical inhibitors of kinases with relevant biological activities to control in vitro and in vivo infections.

Objective 4: We will develop a high-throughput assay to search for inhibitors of the Type 3 secretion systems. We will screen a series of commercial libraries of small molecules for their ability to inhibit type 3 secretion by S. Typhimurium. A systems approach will be employed to understand and characterize the host-pathogen interactions that are manipulated in food animals using novel therapeutic approaches with BT peptides and CpG oligonucleotides without engendering antimicrobial resistance. Microarray analyses of avian and porcine peripheral blood granulocytes and monocytes following treatment with BT peptides or CpG oligonucleotides will be performed. Using InnateDB, bioinformatic interrogation of gene ontology, signaling pathways and transcription factor binding sites will be undertaken; confirmation will be achieved experimentally by qRT-PCR and inhibitor studies of in vitro functional biological assays, and followed up by direct biochemical confirmation. Collectively, these will lead to substantial advances in understanding the complexity of signaling pathways and transcription factors involved in the responses to BT peptides and CpG modulation.

Funding Source:United States Department of Agriculture (USDA), Agricultural Research Service (ARS)
Type:Appropriated
Start Date:2011
End Date:2016
Project Number:6202-32000-031-00
Accession Number:420971
Institutions:USDA/ARS - Southern Plains Area
Investigators:Genovese, Kenneth
He, Haiqi
Kogut, Michael
Swaggerty, Christina
Project Reports:2013 Annual Report
2012 Annual Report
2011 Annual Report
Published Journal
Articles USDA
ARS (NP 108):
Fate of Salmonella Senftenberg in broiler chickens evaluated by challenge experiments
Kallapura G, Kogut MH, Morgan MJ, Pumford NR, Bielke LR, Wolfenden AD, Faulkner OB, Latorre JD, Menconi A, Hernandez-Velasco X, Kuttappan VA, Hargis BM, Tellez G.
Avian Pathol. 2014 Jun 17. [Epub ahead of print]
Selection for pro-inflammatory mediators yields chickens with increased resistance against Salmonella enterica serovar enteritidis
Swaggerty CL, Pevzner IY, Kogut MH.
Poult Sci. 2014 Mar;93(3):535-44.
Obligate brood parasites show more functionally effective innate immune responses: An eco-immunological hypothesis
Hahn DC, Summers S, Genovese KJ, He LH, Kogut MH.
J Evol Biol. 2013 Dec;40(4):554-61.
Combined CpG and poly I:C stimulation of monocytes results in unique signaling activation not observed with the individual ligands
Arsenault RJ, Kogut MH, He H.
Cell Signal. 2013 Nov;25(11):2246-54.
What's so special about chicken immunology? Preface
Lowenthal JW, Bean AG, Kogut MH.
Dev Comp Immunol. 2013 Nov;41(3):307-9.
The avian heterophil
Genovese KJ, He H, Swaggerty CL, Kogut MH.
Dev Comp Immunol. 2013 Nov;41(3):334-40.
Modulation of chicken intestinal gene expression by small cationic peptides as feed additives during the first week post-hatch
Kogut MH, Genovese KJ, He H, Swaggerty CL, Jiang Y.
Clin Vaccine Immunol. 2013 Sep;20(9):1440-8.
The gut microbiota and host innate immunity: Roles in host metabolism and metablic diseases
Kogut MH.
J Appl Poult Res. 2013 Sep;22(3):637-46.
Modulation of chicken intestinal immune gene expression by small cationic peptides as feed additives during the first week posthatch
Kogut MH, Genovese KJ, He H, Swaggerty CL, Jiang Y.
Clin Vaccine Immunol. 2013 Sep;20(9):1440-8.
Enhanced innate immune responses in a brood parasitic cowbird species: Degranulation and oxidative burst
Hahn DC, Summers SG, Genovese KJ, He H, Kogut MH.
Avian Dis. 2013 Jun;57(2):285-9.
Salmonella enterica Typhimurium infection causes metabolic changes in chicken muscle involving AMPK, fatty acid and insulin/mTOR signaling
Arsenault RJ, Napper S, Kogut MH .
Vet Res. 2013 May 17;44:35.
Nitric oxide as a biomarker of intracellular Salmonella viability and identification of the bacteriostatic activity of protein kinase A inhibitor H-89
He LH, Genovese KJ, Swaggerty CL, Nisbet DJ, Kogut MH.
PLoS One. 2013 Mar 15;8(3):e58873.
A comparative study on invasion, survival, modulation of oxidative burst, and nitric oxide responses of macrophages (HD11), and systemic infection in chickens by prevalent poultry Salmonella serovars
He H, Genovese KJ, Swaggerty CL, Nisbet DJ, Kogut MH.
Foodborne Pathog Dis. 2012 Dec;9(12):1104-10.
Gene Expression Analysis of Toll-Like Receptor Pathways in Heterophils from Genetic Chicken Lines that Differ in Their Susceptibility to Salmonella enteritidis
Kogut MH, Chiang HI, Swaggerty CL, Pevzner IY, Zhou H.
Front Genet. 2012 Jul 4;3:121.
Loxoribine pretreatment reduces Salmonella Enteritidis organ invasion in 1-day-old chickens
Swaggerty CL, He H, Genovese KJ, Duke SE, Kogut MH.
Poult Sci. 2012 Apr;91(4):1038-42.
Co-stimulation with TLR3 and TLR21 ligands synergistically up-regulates Th1-cytokine IFN-γ and regulatory cytokine IL-10 expression in chicken monocytes
He H, Genovese KJ, Swaggerty CL, MacKinnon KM, Kogut MH.
Dev Comp Immunol. 2012 Apr;36(4):756-60.
Neutrophil function of neonatal foals is enhanced in vitro by CpG oligodeoxynucleotide stimulation
Bordin AI, Liu M, Nerren JR, Buntain SL, Brake CN, Kogut MH, Cohen ND.
Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):290-7.
Full text available in the NAL Digital Collections.
Bacterial toll-like receptor agonists induce sequential NF-κB-mediated leukotriene B4 and prostaglandin E2 production in chicken heterophils
Kogut MH, He H, Genovese KJ.
Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):159-70.
BT cationic peptides: small peptides that modulate innate immune responses of chicken heterophils and monocytes
Kogut MH, Genovese KJ, He H, Swaggerty CL, Jiang YW.
Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):151-8.
Systemic response to Campylobacter jejuni infection by profiling gene transcription in the spleens of two genetic lines of chickens
Li X, Swaggerty CL, Kogut MH, Chiang HI, Wang Y, Genovese KJ, He H, McCarthy FM, Burgess SC, Pevzner IY, Zhou H.
Immunogenetics. 2012 Jan;64(1):59-69.
Effects of avian triggering receptor expressed on myeloid cells (TREM-A1) activation on heterophil functional activities
Kogut MH, Genovese KJ, Nerren JR, He H.
Dev Comp Immunol. 2012 Jan;36(1):157-65.
Chicken-specific peptide arrays for kinome analysis: Flight for the flightless
Arsenault RJ, Kogut MH.
Curr Top Biotechnol. 2012;7:79-89.
Full text available in the NAL Digital Collections.
Electron-beam Irradiation Inactivation of Salmonella: Effects on Innate Immunity and Induction of Protection Against Salmonella enterica serovar Typhimurium Challenge of Chickens
Kogut MH, McReynolds JL, He H, Genovese KJ, Jesudhasan PR, Davidson MA, Cepeda MA, Pillai SD.
Procedia Vaccinol. 2012;6:47-63.
Non-Journal Publications:
Systemic response to host microflora in gnotobiotic pigs - (Abstract Only)
Genovese, K.J., Harvey, R.B., He, L.H., Nisbet, D.J. 2013. Systemic response to host microflora in gnotobiotic pigs. The Gut Microbiota: The Effector/Regulatory Immune Network. February 10-15, Taos, NM. p. 52(1030).
Food Safety Categories:Methodology
Pathogen Biology
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