interleukin-8 and Enterobacteriaceae-Infections

A new type of CXCL8, named CXCL8_L1b, was identified in this research. Comparison of amino acid sequences of Japanese flounder CXCL8_L1b and CXCL8_L1a (BAB86884.1) showed only 41.2% identity. Transcripts of CXCL8_L1a were highly detected in spleen, kidney, gill and liver, while transcripts of CXCL8_L1b only were detected highly in spleen and kidney of apparently healthy fish. In fish challenged with E. tarda, transcripts of CXCL8_L1a were significantly increased at day 6, while no significant increase was detected in the mRNA level of CXCL8_L1b. On the other hand, fish infected by S. iniae significantly increased both transcripts of CXCL8_L1a and CXCL8_L1b at days 1 and 3. In VHSV-infected fish, only the transcripts of CXCL8_L1b were significantly induced at day 6. LPS and poly I:C stimulation of PBLs induced a high level of CXCL8_L1a transcripts, while CXCL8_L1b transcripts were significantly increased only post poly I:C treatment. To evaluate the chemotactic activity of CXCL8_L1a and CXCL8_L1b, Japanese flounder were intramuscularly injected with recombinant plasmids pCI-CXCL8_L1a and pCI-CXCL8_L1b. H & E staining showed that injections of both pCI-CXCL8_L1a and pCI-CXCL8_L1b caused strong immune responses in the form of intermuscular cell infiltration and capillary congestion. Injection of pCI-CXCL8_L1a and pCI-CXCL8_L1b significantly induced the expressions of genes related to inflammatory response such as IL-6 and CD8α on day 1 post-injection. The transcripts of IgM only significantly increased on day 7 post-injection of pCI-CXCL8_L1b.

Topics: Amino Acid Sequence; Animals; Chemotaxis; Edwardsiella tarda; Enterobacteriaceae Infections; Fish Proteins; Flounder; Immunity; Immunoglobulin M; Interleukin-6; Interleukin-8; Molecular Sequence Data; Novirhabdovirus; Protein Isoforms; Rhabdoviridae Infections; Sequence Alignment

The prone position (PP) has proven beneficial in patients with severe lung injury subjected to mechanical ventilation (MV), especially in those with lobar involvement. We assessed the impact of PP on unilateral pneumonia in rabbits subjected to MV.. After endobronchial challenge with Enterobacter aerogenes, adult rabbits were subjected to either "adverse" (peak inspiratory pressure = 30 cm H2O, zero end-expiratory pressure; n = 10) or "protective" (tidal volume = 8 ml/kg, 5 cm H2O positive end-expiratory pressure; n = 10) MV and then randomly kept supine or turned to the PP. Pneumonia was assessed 8 h later. Data are presented as median (interquartile range).. Compared with the supine position, PP was associated with significantly lower bacterial concentrations within the infected lung, even if a "protective" MV was applied (5.93 [0.34] vs. 6.66 [0.86] log10 cfu/g, respectively; P = 0.008). Bacterial concentrations in the spleen were also decreased by the PP if the "adverse" MV was used (3.62 [1.74] vs. 6.55 [3.67] log10 cfu/g, respectively; P = 0.038). In addition, the noninfected lung was less severely injured in the PP group. Finally, lung and systemic inflammation as assessed through interleukin-8 and tumor necrosis factor-α measurement was attenuated by the PP.. The PP could be protective if the host is subjected to MV and unilateral bacterial pneumonia. It improves lung injury even if it is utilized after lung injury has occurred and nonprotective ventilation has been administered.

Topics: Animals; Endpoint Determination; Enterobacter aerogenes; Enterobacteriaceae Infections; Hemodynamics; Inflammation; Interleukin-8; Lung; Lung Compliance; Male; Pneumonia, Bacterial; Positive-Pressure Respiration; Prone Position; Pulmonary Gas Exchange; Rabbits; Respiration, Artificial; Supine Position; Tumor Necrosis Factor-alpha

Cronobacter sakazakii is a Gram-negative opportunistic pathogen that causes necrotizing enterocolitis, sepsis, and meningitis in premature infants. The genetic basis of C. sakazakii virulence is poorly understood. In this study, the putative LysR-type transcriptional regulator (LTTR) gene (ESA_01081 homolog) was characterized as a possible regulator for C. sakazakii pathogenesis. An in-frame deletion mutant of the ESA_01081 homolog and its cognate complementation strain were constructed and characterized for pathogenesis (adhesion/invasion potentials to human intestinal cells and in vivo rat pup challenge assay), biofilm formation, resistance to oxidative stress and induction of IL-8 secretion. LTTR-deficient C. sakazakii ATCC 29544 exhibits significantly attenuated phenotypes in all the properties tested, except adhesion. Our data strongly suggest that the putative gpESA_01081 homolog, plays an important regulatory role in diverse biological processes including the virulence of C. sakazakii. This is the first report of a functional global regulator in C. sakazakii.

Topics: Animals; Bacterial Adhesion; Bacterial Proteins; Biofilms; Caco-2 Cells; Computational Biology; Cronobacter sakazakii; Enterobacteriaceae Infections; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genes, Regulator; Genetic Complementation Test; Humans; Interleukin-8; Oxidative Stress; Phenotype; Rats; Rats, Sprague-Dawley; Sequence Deletion; Transcription Factors

Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli are noninvasive attaching and effacing (A/E) bacterial pathogens that cause intestinal inflammation and severe diarrheal disease. These pathogens utilize a type III secretion system to deliver effector proteins into host epithelial cells, modulating diverse cellular functions, including the release of the chemokine interleukin-8 (IL-8). While studies have implicated the effectors NleE (non-locus of enterocyte effacement [LEE]-encoded effector E) and NleH1 in suppressing IL-8 release, by preventing NF-κB nuclear translocation, the impact of these effectors only partially replicates the immunosuppressive actions of wild-type EPEC, suggesting another effector or effectors are involved. Testing an array of EPEC mutants, we identified the non-LEE-encoded effector C (NleC) as also suppressing IL-8 release. Infection by ΔnleC EPEC led to exaggerated IL-8 release from infected Caco-2 and HT-29 epithelial cells. NleC localized to EPEC-induced pedestals, with signaling studies revealing NleC inhibits both NF-κB and p38 mitogen-activated protein kinase (MAPK) activation. Using Citrobacter rodentium, a mouse-adapted A/E bacterium, we found that ΔnleC and wild-type C. rodentium-infected mice carried similar pathogen burdens, yet ΔnleC strain infection led to worsened colitis. Similarly, infection with ΔnleC C. rodentium in a cecal loop model induced significantly greater chemokine responses than infection with wild-type bacteria. These studies thus advance our understanding of how A/E pathogens subvert host inflammatory responses.

Topics: Animals; Bacterial Adhesion; Caco-2 Cells; Chemokines; Citrobacter rodentium; Colitis; Enterobacteriaceae Infections; Enteropathogenic Escherichia coli; Epithelial Cells; Escherichia coli Proteins; Fluorescent Antibody Technique; HT29 Cells; Humans; Interleukin-8; Mice; Mice, Inbred C57BL; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Polymerase Chain Reaction

Enteropathogenic Escherichia coli (EPEC) and the murine pathogen Citrobacter rodentium belong to the attaching and effacing (A/E) family of bacterial pathogens. These noninvasive bacteria infect intestinal enterocytes using a type 3 secretion system (T3SS), leading to diarrheal disease and intestinal inflammation. While flagellin, the secreted product of the EPEC fliC gene, causes the release of interleukin 8 (IL-8) from epithelial cells, it is unclear whether A/E bacteria also trigger epithelial inflammatory responses that are FliC independent. The aims of this study were to characterize the FliC dependence or independence of epithelial inflammatory responses to direct infection by EPEC or C. rodentium. Following infection of Caco-2 intestinal epithelial cells by wild-type and DeltafliC EPEC, a rapid activation of several proinflammatory genes, including those encoding IL-8, monocyte chemoattractant protein 1, macrophage inflammatory protein 3alpha (MIP3alpha), and beta-defensin 2, occurred in a FliC-dependent manner. These responses were accompanied by mitogen-activated protein kinase activation, as well as the Toll-like receptor 5 (TLR5)-dependent activation of NF-kappaB. At later infection time points, a subset of these proinflammatory genes (IL-8 and MIP3alpha) was also induced in cells infected with DeltafliC EPEC. The nonmotile A/E pathogen C. rodentium also triggered similar innate responses through a TLR5-independent but partially NF-kappaB-dependent mechanism. Moreover, the EPEC FliC-independent responses were increased in the absence of the locus of enterocyte effacement-encoded T3SS, suggesting that translocated bacterial effectors suppress rather than cause the FliC-independent inflammatory response. Thus, we demonstrate that infection of intestinal epithelial cells by A/E pathogens can trigger an array of proinflammatory responses from epithelial cells through both FliC-dependent and -independent pathways, expanding our understanding of the innate epithelial response to infection by these pathogens.

Topics: Animals; Caco-2 Cells; Chemokine CCL20; Citrobacter rodentium; Dendritic Cells; Enterobacteriaceae Infections; Enteropathogenic Escherichia coli; Epithelial Cells; Escherichia coli Proteins; Flagellin; Gene Expression Regulation; Humans; Inflammation; Interleukin-8; Mice; Mice, Inbred C3H; Mutation; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Time Factors; Toll-Like Receptor 5

Chemokines are important mediators for innate immunity involved in recruitment, activation and adhesion of a variety of leukocyte types to inflammatory foci. While almost all chemokines have been identified from mammals, only a handful of fish chemokines have been identified. Here we report molecular cloning, sequence analysis, and expression of a channel catfish gene resembling interleukin-8 (IL-8). The gene has two alternatively spliced transcripts encoding 114 and 111 amino acids, respectively. The gene has four exons and three introns, typical of the CXC chemokine gene organization. In spite of the structural conservation through evolution, the piscine IL-8 genes showed a much greater sequence divergence than their counterparts among mammals. RT-PCR indicated that both spliced forms were expressed. Expression of the IL-8 like gene was up-regulated 3-5-fold in channel catfish and blue catfish after infection with pathogenic bacteria Edwardsiella ictaluri.

Topics: Animals; Base Sequence; Catfishes; Cloning, Molecular; DNA, Complementary; Edwardsiella ictaluri; Enterobacteriaceae Infections; Gene Expression; Humans; Interleukin-8; Molecular Sequence Data; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Nucleic Acid

The involvement of the proinflammatory cytokines, interleukin 8 (IL-8) and 6 (IL-6), was studied during the first 72 h of acute invasive gastroenteritis. Study population included 33 infants and young children aged six months to six years and seven age-matched controls. As a group, patients with acute invasive gastroenteritis had an increased serum level of IL-8 and IL-6 as compared with healthy controls (p < 0.002 and p < 0.001, respectively). Subjects were then divided into two groups based on stool cultures (proven and non-proven bacterial cultures). Patients with bacterial-proven acute invasive gastroenteritis tended to have increased IL-8 serum concentrations (p < 0.07) as compared with those with non-proven bacterial etiologies and IL-6 levels were only detected in subjects with positive bacterial cultures (p < 0.05). When dividing each sub-group into early and late blood drawing with respect to disease onset, no statistical differences were found in each group but subjects with bacterial-proven etiologies had significant higher IL-6 levels as compared with non-proven etiologies at the two time points (p < 0.019 and p < 0.015, respectively). In conclusion, the proinflammatory cytokines, IL-6 and IL-8, are involved in acute invasive gastroenteritis. The difference in IL-6, and to a lesser degree IL-8, between proven and non-proven bacterial etiologies, needs further investigation.

Topics: Child; Child, Preschool; Enterobacteriaceae; Enterobacteriaceae Infections; Gastroenteritis; Humans; Infant; Interleukin-6; Interleukin-8; Leukocyte Count; Neutrophils; Time Factors

Urinary tract epithelial cells (T 24/83) are able to express interleukin (IL)-6, IL-8, platelet-derived growth factor (PDGF) and tumour necrosis factor-alpha, but not IL-1 beta, IL-2, IL-4 and IL-10 in response to an infection with uropathogenic bacteria. The process of cytokine secretion is time dependent, with a significant increase in the cytokine activity after 60 min. The expression of virulence factors of the bacteria does not seem to play a role. The interaction between bacterial products (e.g. lipopolysaccharide) and/or bacterial adhesion mediated by adhesins and specific receptor molecules of cell surfaces may be responsible for the activity of mediator protein expression in the epithelial cells. The release of PDGF and IL-8 was found to be higher when due to Escherichia coli HB 101 (rough form) than that caused by other bacterial strains. Citrobacter CB 3009 provoked the highest level of IL-6. The PDGF level correlated significantly with IL-6 and IL-8 values (P<0.001). There was a significant correlation between the time-dependent release of IL-6 and IL-8 (P<0.05). In epithelial cytokine response to bacterial infection, the reaction of the epithelial cells may modify themselves (e.g. internalization of bacteria) and the immuno-regulatory processes that are caused by infection and responsible for parenchymal injury.

Topics: Citrobacter; Cytokines; Enterobacteriaceae Infections; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Escherichia coli Infections; Humans; Interleukin-6; Interleukin-8; Platelet-Derived Growth Factor; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha; Urinary Bladder Neoplasms; Urinary Tract Infections; Urogenital System; Urothelium; Virulence

Enteropathogenic Escherichia coli (EPEC) is an extracellular bacterial pathogen that infects the human intestinal epithelium and is a major cause of infantile diarrhea in developing countries. EPEC belongs to the group of attaching and effacing (A/E) pathogens. It uses a type III secretion system to deliver proteins into the host cell that mediate signal transduction events in host cells. We used gene array technology to study epithelial cell responses to EPEC infection at the level of gene expression. We found that EPEC induces the expression of several genes in infected HeLa cells by a lipopolysaccharide (LPS)-independent mechanism, including cytokines and early growth response factor 1 (Egr-1). The transcription factor Egr-1 is an immediate-early-induced gene that is activated in most cell types in response to stress. EPEC-induced upregulation of egr-1 is mediated by the activation of the MEK/extracellular signal-regulated kinase signal transduction pathway and is dependent on the type III secretion system. egr-1 is also induced during infection of mice by the A/E pathogen Citrobacter rodentium, suggesting that both Egr-1 and the activation of this mitogen-activated protein kinase signal transduction pathway may play a role in disease.

Topics: Animals; Blotting, Northern; Citrobacter freundii; DNA-Binding Proteins; Early Growth Response Protein 1; Enterobacteriaceae Infections; Enzyme Activation; Epithelial Cells; Escherichia coli Infections; Gene Expression; HeLa Cells; Humans; Immediate-Early Proteins; Interleukin-8; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Phosphorylation; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors

Human intestinal epithelial cells up-regulate the expression of an inflammatory gene program in response to infection with a spectrum of different strains of enteroinvasive bacteria. The conserved nature of this program suggested that diverse signals, which are activated by enteroinvasive bacteria, can be integrated into a common signaling pathway that activates a set of proinflammatory genes in infected host cells. Human intestinal epithelial cell lines, HT-29, Caco-2, and T84, were infected with invasive bacteria that use different strategies to induce their uptake and have different intracellular localizations (i.e., Salmonella dublin, enteroinvasive Escherichia coli, or Yersinia enterocolitica). Infection with each of these bacteria resulted in the activation of TNF receptor associated factors, two recently described serine kinases, I kappa B kinase (IKK) alpha and IKK beta, and increased NF-kappa B DNA binding activity. This was paralleled by partial degradation of I kappa B alpha and I kappa B epsilon in bacteria-infected Caco-2 cells. Mutant proteins that act as superrepressors of IKK beta and I kappa B alpha inhibited the up-regulated transcription and expression of downstream targets genes of NF-kappa B that are key components of the epithelial inflammatory gene program (i.e., IL-8, growth-related oncogene-alpha, monocyte chemoattractant protein-1, TNF-alpha, cyclooxygenase-2, nitric oxide synthase-2, ICAM-1) activated by those enteroinvasive bacteria. These studies position NF-kappa B as a central regulator of the epithelial cell innate immune response to infection with enteroinvasive bacteria.

Topics: Caco-2 Cells; Chemokine CCL2; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; DNA-Binding Proteins; Enterobacteriaceae Infections; Enzyme Activation; Gene Expression Regulation; Gene Expression Regulation, Bacterial; Genes, Reporter; Growth Substances; HT29 Cells; Humans; I-kappa B Kinase; I-kappa B Proteins; Immunity, Innate; Intercellular Adhesion Molecule-1; Intercellular Signaling Peptides and Proteins; Interleukin-8; Intestinal Mucosa; Intracellular Fluid; Isoenzymes; Membrane Proteins; NF-kappa B; NF-KappaB Inhibitor alpha; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Prostaglandin-Endoperoxide Synthases; Protein Serine-Threonine Kinases; Signal Transduction; Tumor Necrosis Factor-alpha