Unique genome-wide transcriptome profiles of chicken macrophages exposed to Salmonella-derived endotoxin

Departments of Animal Science, Iowa State University, Ames, Iowa 50011, USA.
BMC Genomics (Impact Factor: 3.99). 10/2010; 11(1):545. DOI: 10.1186/1471-2164-11-545
Source: PubMed


Macrophages play essential roles in both innate and adaptive immune responses. Bacteria require endotoxin, a complex lipopolysaccharide, for outer membrane permeability and the host interprets endotoxin as a signal to initiate an innate immune response. The focus of this study is kinetic and global transcriptional analysis of the chicken macrophage response to in vitro stimulation with endotoxin from Salmonella typhimurium-798.
The 38535-probeset Affymetrix GeneChip Chicken Genome array was used to profile transcriptional response to endotoxin 1, 2, 4, and 8 hours post stimulation (hps). Using a maximum FDR (False Discovery Rate) of 0.05 to declare genes as differentially expressed (DE), we found 13, 33, 1761 and 61 DE genes between endotoxin-stimulated versus non-stimulated cells at 1, 2, 4 and 8 hps, respectively. QPCR demonstrated that endotoxin exposure significantly affected the mRNA expression of IL1B, IL6, IL8, and TLR15, but not IL10 and IFNG in HD 11 cells. Ingenuity Pathway Analysis showed that 10% of the total DE genes were involved in inflammatory response. Three, 9.7, 96.8, and 11.8% of the total DE inflammatory response genes were significantly differentially expressed with endotoxin stimulation at 1, 2, 4 and 8 hps, respectively. The NFKBIA, IL1B, IL8 and CCL4 genes were consistently induced at all times after endotoxin treatment. NLRC5 (CARD domain containing, NOD-like receptor family, RCJMB04_18i2), an intracellular receptor, was induced in HD11 cells treated with endotoxin.
As above using an in vitro model of chicken response to endotoxin, our data revealed the kinetics of gene networks involved in host response to endotoxin and extend the known complexity of networks in chicken immune response to Gram-negative bacteria such as Salmonella. The induction of NFKBIA, IL1B, IL8, CCL4 genes is a consistent signature of host response to endotoxin over time. We make the first report of induction of a NOD-like receptor family member in response to Salmonella endotoxin in chicken macrophages.

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Available from: Michael J Wannemuehler, Oct 07, 2015
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    • "Microarray technology is a powerful tool which has been used in the chicken to analyze global gene expression profiles during Salmonella infection. Most of the studies pertain to the serovar Enteritidis, some of these studies were performed in vitro on macrophages or cells from the granulosa (Chiang et al., 2008; Ciraci et al., 2010; Tsai et al., 2010; Zhang et al., 2008). Gene expression profiling was studied in vivo on whole organs such as the jejunum (Schokker et al., 2010, 2012; van Hemert et al., 2007), ceca (Higgins et al., 2011) and spleen (Zhou and Lamont, 2007) during the first days of infection. "
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    ABSTRACT: Infection of chicken with Salmonella may lead to a carrier-state characterized by the persistence of bacteria in the ceca for a long period of time and result in their excretion in feces This excretion is the source of contamination of their congeners and food During infection, enterocytes are the primary target cells for Salmonella, the producers of soluble factors which launch immune response and cells which are reciprocally responsive to surrounding immune cells This study used microarrays to compare the gene expression profile during carrier-state of enterocytes purified from infected and control chicks which are either resistant or susceptible to Salmonella Enteritidis carrier-state In total, we identified 271 genes significantly differentially expressed with an absolute fold change greater than 15 A global analysis determined interaction networks between differentially regulated genes Using an a priori approach, our analyses focused on differentially expressed genes which were transcriptionally linked to cytokines playing a major role in the fate of the immune response The expression of genes transcriptionally linked to type I interferon and TGF-β was down-regulated in infected chicks from both lines Gene expression linked to the Th1 axis suggests the latter is inhibited in both lines Finally, the expression of genes linked to IL-4, IL-5 and IL-13 indicates that susceptibility to carrier-state could be associated with a Th2 bias Overall, these results highlight that the response to Salmonella during the acute phase and carrier-state is different and that enterocytes play a central role in this response
    Veterinary Immunology and Immunopathology 05/2014; 159(1). DOI:10.1016/j.vetimm.2014.03.001 · 1.54 Impact Factor
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    • "NLRC5 is a positive IFN mediator in response to viral infection [96], while it attenuates the antiviral response to VSV [97]. In chickens, the inhibitory effects of NLRC5 on inflammatory pathways are well understood [98,99]. The expression of chicken NLRC5 is greatly increased in LPS-treated HD11 cells, but not in poly(I:C)-treated HD11 cells. "
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    ABSTRACT: : Similar to mammals, several viral-sensing pattern recognition receptors (PRR) have been identified in birds including Toll-like receptors (TLR) and retinoic acid-inducible gene I (RIG-I)-like receptors (RLR). Avian TLR are slightly different from their mammalian counterparts, including the pseudogene TLR8, the absence of TLR9, and the presence of TLR1La, TLR1Lb, TLR15, and TLR21. Avian TLR3 and TLR7 are involved in RNA virus recognition, especially highly pathogenic avian influenza virus (HPAIV), while TLR15 and TLR21 are potential sensors that recognize both RNA viruses and bacteria. However, the agonist of TLR15 is still unknown. Interestingly, chickens, unlike ducks, geese and finches, lack RIG-I, however they do express melanoma differentiation-associated gene 5 (MDA5) which functionally compensates for the absence of RIG-I. Duck RIG-I is the cytosolic recognition element for HPAIV recognition, while chicken cells sense HPAIV through MDA5. However, the contributions of MDA5 and RIG-I to IFN-beta induction upon HPAIV infection is different, and this may contribute to the chicken's susceptibility to highly pathogenic influenza. It is noteworthy that the interactions between avian DNA viruses and PRR have not yet been reported. Furthermore, the role for avian Nod-like receptors (NLR) in viral immunity is largely unknown. In this review, recent advances in the field of viral recognition by different types of PRR in birds are summarized. In particular, the tissue and cellular distribution of avian PRR, the recognition and activation of PRR by viruses, and the subsequent expression of innate antiviral genes such as type I IFN and proinflammatory cytokines are discussed.
    Veterinary Research 09/2013; 44(1):82. DOI:10.1186/1297-9716-44-82 · 2.82 Impact Factor
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    • "Although the number (44) of genes differentially expressed at a significance level of q value <0.05 in this study was low, it is similar to that found in some related studies on response to bacteria or bacterial components in chicken tissues or cells. In an in vitro study in which chicken macrophage cells were stimulated with Salmonella endotoxin, Ciraci et al. (2010) reported that only 13 genes were differentially expressed at a significance level of q value < 0.05 at 1-h poststimulation and that 33 genes were differentially expressed at 2-h poststimulation . In the same study, 1,761 genes were differentially expressed at 4-h poststimulation. "
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    ABSTRACT: Salmonella enterica serovar Enteritidis is an enteric bacterium that can contaminate chicken eggs and meat, resulting in production losses and consumer illness. To provide insight into the systemic metabolic effects of S. Enteritidis infection, liver samples were harvested 10 days post infection from broiler hens. Hepatic global gene expression levels were assessed using a chicken 44K Agilent microarray. Forty-four genes were differentially expressed at a significance level of q-value < 0.05. One hundred eighty-three genes were differentially expressed at a suggestive significance level of q-value < 0.1. A predominance of down-regulation existed among significantly differentially expressed genes. Cell cycle and metabolism networks were created from the differentially expressed genes. Mitochondria-mediated apoptosis, electron transport, peptidase activity, vein constriction, cell differentiation, IL-2 signaling, Jak-Stat signaling, B-cell receptor signaling, GDP/GTP exchange, and protein recycling were among the functions of the differentially expressed genes that were down-regulated in response to S. Enteritidis. The effects of S. Enteritidis infection on the liver transcriptome profiles of broilers reflect a predominance of down-regulation of genes with cell cycle and metabolic functions. The most pronounced response was the down-regulation of genes that function in metabolic pathways, inflammation, and mitochondria-mediated apoptosis. These results provide insight into important systemic metabolic mechanisms that are active in the chicken liver in response to S. Enteritidis infection at 10 days post infection. © 2012 Wiley Periodicals, Inc.
    genesis 05/2013; 51(5). DOI:10.1002/dvg.22351 · 2.02 Impact Factor
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