interleukin-8 and Enterocolitis--Pseudomembranous

The interleukin 8 gene single-nucleotide polymorphism rs4073/-251T >A predisposes to Clostridium difficile infection (CDI), but this association has not been independently validated. In this study, we were unable to replicate this association in either a white cohort or by meta-analysis, suggesting that rs4073/-251T >A is unlikely to constitute a major risk factor for CDI.

Topics: Adult; Aged; Anti-Bacterial Agents; Case-Control Studies; Clostridioides difficile; Diarrhea; Enterocolitis, Pseudomembranous; Feces; Female; Genetic Predisposition to Disease; Humans; Interleukin-8; Male; Polymorphism, Single Nucleotide; Promoter Regions, Genetic

Topics: Bacterial Toxins; Cholera; Cyclic AMP; Diarrhea; Enterocolitis, Pseudomembranous; Escherichia coli Infections; Guanylate Cyclase; Humans; Interleukin-8; Nutrition Disorders; Platelet Activating Factor; Population Growth; Signal Transduction

Clostridium difficile is the most common etiological agent of antibiotic-associated diarrhea in hospitalized and non-hospitalized patients. This study investigated the secretion of membrane vesicles (MVs) from C. difficile and determined the expression of pro-inflammatory cytokine genes and cytotoxicity of C. difficile MVs in epithelial cells in vitro. C. difficile ATCC 43255 and two clinical isolates secreted spherical MVs during in vitro culture. Proteomic analysis revealed that MVs of C. difficile ATCC 43255 contained a total of 262 proteins. Translation-associated proteins were the most commonly identified in C. difficile MVs, whereas TcdA and TcdB toxins were not detected. C. difficile ATCC 43255-derived MVs stimulated the expression of pro-inflammatory cytokine genes, including interleukin (IL)-1β, IL-6, IL-8, and monocyte chemoattractant protein-1 in human colorectal epithelial Caco-2 cells. Moreover, these extracellular vesicles induced cytotoxicity in Caco-2 cells. In conclusion, C. difficile MVs are important nanocomplexes that elicit a pro-inflammatory response and induce cytotoxicity in colonic epithelial cells, which may contribute, along with toxins, to intestinal mucosal injury during C. difficile infection.

Topics: Bacterial Proteins; Bacterial Toxins; Caco-2 Cells; Cell Culture Techniques; Chemokine CCL2; Clostridioides difficile; Colon; Cytokines; Enterocolitis, Pseudomembranous; Enterotoxins; Epithelial Cells; Flow Cytometry; Gene Expression Regulation; Hep G2 Cells; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Intestinal Mucosa; Microscopy, Electron, Transmission; Proteomics

Clostridium difficile infection (CDI) is the most common cause of infectious diarrhea and represents an important burden for healthcare worldwide. Symptoms of severe CDI include watery, foul-smelling diarrhea, peripheral leucocytosis, increased C-reactive protein (CRP), acute renal failure, hypotension and pseudomembranous colitis. Recent studies indicate that the main cause of CDI is dysbiosis, an imbalance in the normal gut microbiota. The restoration of a healthy gut microbiota composition via fecal microbiota transplantation (FMT) recently became more popular. The aim of the present study was to assess the effect of FMT on the healing of CDI and to analyze the changes in the level of pro-inflammatory markers (C-reactive protein, fecal calprotectin) and pro-inflammatory cytokines. Eighteen patients with CDI were included in our study (6 males and 12 females) with recurrent and/or severe CDI. The FMT was performed in 17 patients using colonoscopy, including 16 patients receiving a one-time FMT and 1 patient who needed 2 additional FMTs. One patient was treated with a single round of FMT using push-and-pull enteroscopy. In all CDI patients, before and 3 weeks after FMT, the following parameters were analyzed: C-reactive protein, fecal calprotectin, and plasma interleukin (IL)-6, IL-8 and IL-12, and tumor necrosis factor-alpha (TNF-α). In addition, the plasma level of LL-37, a cathelicidine peptide was assessed by fluorescence-activated cell sorting (FACS) before and 3 months after FMT. Finally, in 7 patients a microbiome analysis was performed by sequencing of 16SrRNA in stool probes obtained before and 3 weeks after FMT. The healing rate of CDI was 94%. In all successfully treated patients no recurrent CDI was observed during follow-up (16 months). The serum level of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-8 and IL-12) significantly decreased after FMT. Similarly, CRP and fecal calprotectin normalized after FMT. 3 months after FMT a significant increase of LL-37 in the plasma of successfully treated patients was monitored. The sequencing analysis demonstrated an elevated abundance of beneficial bacterial species such as Lactobacillaceae, Ruminococcaceae, Desulfovibrionaceae, Sutterellaceae and Porphyromonodacea after FMT. No serious side effects were observed. We concluded that FMT represented a very effective and safe treatment of recurrent and/or severe CDI and led to favorable shifts in the composition of gut microbiome.

Topics: Aged; Anti-Bacterial Agents; C-Reactive Protein; Clostridioides difficile; Clostridium Infections; Colonoscopy; Diarrhea; Enterocolitis, Pseudomembranous; Fecal Microbiota Transplantation; Feces; Female; Gastrointestinal Microbiome; Humans; Inflammation; Interleukin-6; Interleukin-8; Male; Treatment Outcome; Tumor Necrosis Factor-alpha

The efficacy of oral tigecycline treatment (2 mg/kg of body weight for 7 days) of Clostridium difficile infection (CDI) was evaluated in the gnotobiotic pig model, and its effect on human gut microflora transplanted into the gnotobiotic pig was determined. Tigecycline oral treatment improved survival, clinical signs, and lesion severity and markedly decreased concentrations of Firmicutes but did not promote CDI. Our data showed that oral tigecycline treatment has a potential beneficial effect on the treatment of CDI.

Topics: Administration, Oral; Animals; Anti-Bacterial Agents; Clostridioides difficile; Drug Administration Schedule; Enterocolitis, Pseudomembranous; Fluoroquinolones; Germ-Free Life; Humans; Interleukin-8; Microbiota; Minocycline; Pyrimidinones; Swine; Tigecycline; Vancomycin

To identify specific fecal biomarkers for symptomatic Clostridium difficile infection and predictors of poor outcomes.. We enrolled 65 children with positive C difficile testing (cases) and 37 symptomatic controls. We also analyzed stool samples from colonized and non-colonized asymptomatic children. We performed enzyme immunoassays to determine fecal interleukin (IL)-8, lactoferrin, and phosphorylated-p38 protein concentrations, and quantitative polymerase chain reaction to determine IL-8 and chemokine ligand (CXCL)-5 RNA relative transcript abundances, and C difficile bacterial burden.. Of 68 asymptomatic controls, 16 were colonized with C difficile. Phosphorylated-p38 was specific for C difficile infection but lacked sensitivity. Fecal cytokines were elevated in samples from symptomatic children, whether cases or controls. In children with C difficile infection, fecal CXCL-5 and IL-8 messenger RNA abundances at diagnosis correlated with persistent diarrhea after 5 days of C difficile infection therapy and with treatment with vancomycin. When children with concomitant viral gastroenteritis were excluded, these correlations persisted. Time-to-diarrhea resolution was significantly longer in patients with elevated fecal cytokines at diagnosis. A logistic regression model identified high CXCL-5 messenger RNA abundance as the only predictor of persistent diarrhea. Conversely, fecal C difficile bacterial burden was not different in symptomatic and asymptomatic children and did not correlate with any clinical outcome measure.. Fecal inflammatory cytokines may be useful in distinguishing C difficile colonization from disease and identifying children with C difficile infection likely to have prolonged diarrhea.

Topics: Biomarkers; Case-Control Studies; Child; Enterocolitis, Pseudomembranous; Feces; Female; Humans; Interleukin-8; Lactoferrin; Male; p38 Mitogen-Activated Protein Kinases; Prospective Studies

C. difficile is a Gram-positive spore-forming anaerobic bacterium that is the leading cause of nosocomial diarrhea in the developed world. The pathogenesis of C. difficile infections (CDI) is driven by toxin A (TcdA) and toxin B (TcdB), secreted factors that trigger the release of inflammatory mediators and contribute to disruption of the intestinal epithelial barrier. Neutrophils play a key role in the inflammatory response and the induction of pseudomembranous colitis in CDI. TcdA and TcdB alter cytoskeletal signaling and trigger the release of CXCL8/IL-8, a potent neutrophil chemoattractant, from intestinal epithelial cells; however, little is known about the surface receptor(s) that mediate these events. In the current study, we sought to assess whether toxin-induced CXCL8/IL-8 release and barrier dysfunction are driven by the activation of the P2Y6 receptor following the release of UDP, a danger signal, from intoxicated Caco-2 cells. Caco-2 cells express a functional P2Y6 receptor and release measurable amounts of UDP upon exposure to TcdA/B. Toxin-induced CXCL8/IL-8 production and release were attenuated in the presence of a selective P2Y6 inhibitor (MRS2578). This was associated with inhibition of TcdA/B-induced activation of NFκB. Blockade of the P2Y6 receptor also attenuated toxin-induced barrier dysfunction in polarized Caco-2 cells. Lastly, pretreating mice with the P2Y6 receptor antagonists (MSR2578) attenuated TcdA/B-induced inflammation and intestinal permeability in an intrarectal toxin exposure model. Taken together these data outline a novel role for the P2Y6 receptor in the induction of CXCL8/IL-8 production and barrier dysfunction in response to C. difficile toxin exposure and may provide a new therapeutic target for the treatment of CDI.

Topics: Animals; Apyrase; Caco-2 Cells; Clostridioides difficile; Disease Models, Animal; Enterocolitis, Pseudomembranous; Enterotoxins; Humans; Inflammation; Interleukin-8; Intestinal Mucosa; Male; Mice; NF-kappa B; Purinergic P2 Receptor Antagonists; Receptors, Purinergic P2; Signal Transduction

Neutrophil recruitment coordinated by intestinal interleukin (IL)-8 secretion is a key component in the pathogenesis of Clostridium difficile infection (CDI). We hypothesized that a common single-nucleotide polymorphism (SNP) in the -251 region of the IL-8 gene promoter may be predictive of recurrent CDI.. This was a prospective cohort study of hospitalized adult patients with CDI who were admitted to a large, university-affiliated medical center from 2007 through 2008. Patients were monitored for 3 months after diagnosis of CDI and assessed for recurrent CDI (defined as a return of diarrhea that required treatment after initial symptom resolution). DNA was isolated from blood samples, and genetic sequencing was performed using polymerase chain reaction and pyrosequencing. The association between IL-8 genotype and recurrent CDI was assessed using univariate and multivariate statistics.. Ninety-six patients with a mean (± standard deviation) age of 61 ± 16 years (54% of whom were female and 63% of whom were white) were identified. The overall incidence of recurrent CDI was 24%. IL-8 allele frequency was similar to previously reported findings (for A/A, 27%; for A/T, 53%; and for T/T, 20%). The incidence of recurrent CDI was 38% in patients with the A/A allele and 19% in all other patients (relative risk, 2.1; 95% confidence interval, 1.04-4.13) (P = .043).. This study indicates that a common SNP in the IL-8 gene promoter is an independent predictor of recurrent CDI. Our results could offer risk stratification for patients at high risk for recurrent CDI.

Topics: Aged; Clostridioides difficile; Cohort Studies; Enterocolitis, Pseudomembranous; Female; Gene Frequency; Genetic Predisposition to Disease; Hospitalization; Humans; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Prospective Studies; Recurrence; Sequence Analysis, DNA

This study was undertaken to evaluate the gene expression profile in monocytes from three patients with reactive arthritis (ReA) in remission in order to identify candidate genes accounting for a potential susceptibility to ReA. Gene expression analyses revealed eight differentially expressed mRNA transcripts in monocytes of ReA patients. The major part of genes encoded cytokines, growth factors and chemokines. There was a remarkably high proportion of proangiogenic factors, in particular IP10, ENA-78, and IL-8 accounting for a genetically determined susceptibility to ReA at the host cell level.

Topics: Arthritis, Reactive; Case-Control Studies; Chemokine CXCL5; Chlamydia Infections; Chlamydia trachomatis; Clostridioides difficile; Enterocolitis, Pseudomembranous; Gene Expression Profiling; Genetic Predisposition to Disease; Humans; Interleukin-8; Prohibitins; Receptors, Cytokine; RNA, Messenger; Yersinia enterocolitica; Yersinia Infections

Topics: Aged, 80 and over; Anti-Bacterial Agents; Enterocolitis, Pseudomembranous; Erythromycin; Humans; Interleukin-8

Previous studies have shown that failure to produce serum antibodies to C. difficile (CD) toxin A is associated with more severe and recurrent C. difficile-associated diarrhea (CDAD); and that presence of AA genotype in the interleukin (IL)-8 gene promoter -251 position is associated with increased susceptibility to CDAD. This study examined the relationship between serum immunoglobulin G antibodies to CD toxin A and the presence of IL-8 AA genotype in hospitalized patients with CDAD.. At enrollment, blood for host IL-8 genotype, serum for CD anti-toxin A antibody, and stool for IL-8 by enzyme-linked immunosorbent assay were obtained in CDAD patients and in CD-toxin-negative asymptomatic controls.. Nine of 24 (37.5%) CDAD and 3 of 20 (15%) controls were CD anti-toxin A positive (P = .095). Eleven of 24 (45.8%) CDAD subjects were positive for AA genotype compared with 5 of 20 (25.0%) controls (P = .0019). One of 11 (9.1%) CDAD with AA genotype were positive for anti-toxin A antibodies compared with 8 of 13 (61.5%) non-AA genotype CDAD (P < .0001). Fecal IL-8 concentration for the single antibody-positive CDAD subject with AA genotype was lower than the median level of 822 microg/mL seen in 10 anti-toxin A antibody-negative subjects with CDAD.. This study provided evidence that host susceptibility to C. difficile diarrhea is related both to a defective humoral immune response to CD toxin A and host IL-8 AA genotype.

Topics: Aged; Antibodies, Anti-Idiotypic; Bacterial Toxins; Clostridioides difficile; Diarrhea; DNA; Enterocolitis, Pseudomembranous; Enterotoxins; Enzyme-Linked Immunosorbent Assay; Follow-Up Studies; Glucosyltransferases; Humans; Immunoglobulin G; Interleukin-8; Middle Aged; Odds Ratio; Phenotype; Polymerase Chain Reaction; Polymorphism, Genetic; Prognosis; Retrospective Studies

Mucosal interleukin 8 (IL-8) and neutrophil recruitment are central to the pathogenesis of Clostridium difficile (CD) toxin-induced diarrhea (CDD). We hypothesized that like other inflammatory mucosal infections, susceptibility to CDD would relate to genetically determined variations in the production of IL-8.. Fecal IL-8 production and single nucleotide polymorphism (SNP) frequency in the -251 region of the IL-8 gene were determined in hospitalized patients: 42 with CDD, 42 with CD-negative diarrhea, and 41 without diarrhea. Cases and controls were matched by age, length of hospital stay, comorbidity, and receipt of antibiotics.. An association was found between the IL-8 -251 A/A allele and occurrence of CDD, 39%versus 16% (OR = 3.26, 95% CI 1.09-9.17) and 17% (OR = 5.50, 95% CI 1.22-24.8) for the two control groups. Comparing results by IL-8 genotype for the CDD cases, median and mean fecal IL-8 levels were significantly higher for the -251 A/A genotype (p = 0.03 for median and 0.001 for mean).. These studies indicate a common SNP in the IL-8 gene is associated with increased susceptibility to CDD and with increased fecal IL-8 in diarrheal stools.

Topics: Clostridioides difficile; Diarrhea; Enterocolitis, Pseudomembranous; Feces; Female; Humans; Interleukin-8; Male; Middle Aged; Polymorphism, Genetic; Polymorphism, Single Nucleotide; Promoter Regions, Genetic

Clostridium difficile colitis causes striking leukocytosis. We examined the possibility that toxins A or B, or other nontoxin products of C. difficile, act as superantigens, thereby stimulating leukocytosis. Our results failed to show major histocompatibility complex class II-dependent T lymphocyte proliferation, the hallmark of superantigen activity. Elevated white blood cell counts in C. difficile colitis are probably due to increased generation of cytokines such as interleukin-6 (IL-6) or IL-8.

Topics: Animals; Bacterial Proteins; Bacterial Toxins; Clostridioides difficile; Enterocolitis, Pseudomembranous; Enterotoxins; Humans; Interleukin-6; Interleukin-8; Leukocytosis; Mice; Superantigens

Clostridium difficile toxin A causes acute neutrophil infiltration and intestinal mucosal injury. In cultured cells, toxin A inactivates Rho proteins by monoglucosylation. In monocytes, toxin A induces IL-8 production and necrosis by unknown mechanisms. We investigated the role of mitogen-activated protein (MAP) kinases in these events. In THP-1 monocytic cells, toxin A activated the 3 main MAP kinase cascades within 1 to 2 minutes. Activation of p38 was sustained, whereas stimulation of extracellular signal-regulated kinases and c-Jun NH(2)-terminal kinase was transient. Rho glucosylation became evident after 15 minutes. IL-8 gene expression was reduced by 70% by the MEK inhibitor PD98059 and abrogated by the p38 inhibitor SB203580 or by overexpression of dominant-negative mutants of the p38-activating kinases MKK3 and MKK6. SB203580 also blocked monocyte necrosis and IL-1beta release caused by toxin A but not by other toxins. Finally, in mouse ileum, SB203580 prevented toxin A-induced neutrophil recruitment by 92% and villous destruction by 90%. Thus, in monocytes exposed to toxin A, MAP kinase activation appears to precede Rho glucosylation and is required for IL-8 transcription and cell necrosis. p38 MAP kinase also mediates intestinal inflammation and mucosal damage induced by toxin A.

Topics: Animals; Bacterial Toxins; Cell Line; Clostridioides difficile; Enteritis; Enterocolitis, Pseudomembranous; Enterotoxins; Enzyme Activation; Gene Expression Regulation; Glycosylation; Interleukin-8; MAP Kinase Signaling System; Mice; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinases; Monocytes; Neutrophil Infiltration; Neutrophils; p38 Mitogen-Activated Protein Kinases; rho GTP-Binding Proteins

Twenty-two patients with Clostridium difficile colitis as determined by positive enzyme immunoassay for toxin A were evaluated for fecal inflammatory markers and their relationship to the severity of illness. Fourteen of 22 specimens were positive for fecal lactoferrin (FLF), with titers from 1:50 to 1:800. Nine of 10 stools tested had ratios of interleukin-1beta (IL-1beta) to IL-1 receptor antagonist (IL-1ra) of >0.01. Seventeen of 22 specimens also had elevated IL-8 concentrations, and 12 of 14 had elevated IL-1beta concentrations. A review of the 18 available patient records revealed that fecal IL-8 concentrations, IL-1beta/IL-1ra ratios, and FLF titers were significantly higher in patients with moderate to severe disease than in patients with mild disease. These findings suggest that the proinflammatory effects of C. difficile may directly influence clinical characteristics of human disease.

Topics: Adult; Aged; Aged, 80 and over; Child; Clostridioides difficile; Enterocolitis, Pseudomembranous; Feces; Female; Humans; Interleukin-1; Interleukin-8; Lactoferrin; Male; Middle Aged; Receptors, Interleukin-1