interleukin-8 and Helicobacter-Infections

The review deals with the current aspects of prevention of non-cardia gastric cancer (GC). Helicobacter pylori is the most common cause of non-cardia GC. The Correa cascade remains a major pattern of the pathogenesis of non-cardia GC as before. The key moments in gastric carcinogenesis are H. pylori infection; genes associated with cell recognition of bacteria; an immune response and the activation of an inflammatory response. The prevention of GC requires H. pylori eradication as primary prevention in combination with screening for this pathology as secondary prevention of gastric malignancies. Standard three-component therapy is a first-line major regimen for H. pylori eradication.

Topics: Antibodies, Bacterial; Drug Therapy, Combination; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-8; Polymorphism, Genetic; Receptors, Interleukin-1; Risk Factors; Stomach Neoplasms

Helicobacter pylori (H. pylori) is a major etiological factor in the development of gastric cancer. Large-scale epidemiological studies have confirmed the strong association between H. pylori infection and both cancer development and progression. Interleukin-8 (IL-8) is overexpressed in gastric mucosa exposed to H. pylori. The expression of IL-8 directly correlates with a poor prognosis in gastric cancer. IL-8 is multifunctional. In addition to its potent chemotactic activity, it can induce proliferation and migration of cancer cells. In this review, we focus on recent insights into the mechanisms of IL-8 signaling associated with gastric cancer. The relationship between IL-8 and H. pylori is discussed. We also summarize the current therapeutics against IL-8 in gastric cancer.

Topics: Animals; Antineoplastic Agents; Cell Transformation, Neoplastic; Drug Design; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Molecular Targeted Therapy; Prognosis; Receptors, Interleukin-8; Risk Factors; Signal Transduction; Stomach Neoplasms

It remains controversial regarding the association between interleukin-8 (IL-8) gene -251 T/A polymorphism and peptic ulcer disease (PUD) risk. Thus, a large-scale meta-analysis evaluating the precise association between this gene variant and PUD risk is required. We searched the PubMed, Embase, Web of Science, and Google Scholar until April 25, 2012. Additionally, hand searching of the references of identified articles were performed. All the statistical tests were performed using Stata 11.0. A total of eight studies (3105 subjects) were included in this meta-analysis. Overall, no significant association was found between IL-8 gene -251 T/A polymorphism and PUD risk (for A allele vs. T allele: OR = 1.17, 95% CI = 0.97-1.41, p = 0.094; for A/A vs. T/T: OR = 1.33, 95% CI = 0.94-1.90, p = 0.108; for A/A vs. A/T+T/T: OR = 1.22, 95% CI =0.97-1.52, p = 0.083; for A/A+A/T vs. T/T: OR = 1.26, 95% CI = 0.95-1.67, p = 0.113). However, in the subgroup analyses by ethnicity, H. pylori infection and the subtype of PUD, significant associations were found between IL-8 gene -251 T/A polymorphism and PUD risk in Asians, H. pylori+, duodenal ulcer disease (DUD) and gastric ulcer disease (GUD), respectively. In summary, the present meta-analysis suggests that IL-8 gene -251 T/A polymorphism is associated with increased PUD risk among Asians, and especially for the subgroups of H. pylori+, DUD and GUD.

Topics: Alleles; Asian People; Case-Control Studies; Databases, Bibliographic; Duodenal Ulcer; Gene Frequency; Haplotypes; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Risk; Stomach Ulcer

Potential functional allele A/T single nucleotide polymorphism (SNP) of Interleukin 8 (IL-8) promoter -251 has been implicated in gastric cancer risk.. We aimed to explore the role of A/T SNP of IL-8 -251 in the susceptibility to gastric cancer through a systematic review and meta-analysis. Each initially included article was scored for quality appraisal. Desirable data were extracted and registered into databases. Eighteen studies were ultimately eligible for the meta-analysis of IL-8 - 251 A/T SNP. We adopted the most probably appropriate genetic model (codominant model). Potential sources of heterogeneity were sought out via stratification and sensitivity analyses, and publication biases were estimated.. Between IL-8 -251 AA genotype with gastric cancer risk, statistically significant association could be noted with overall gastric cancer, evidently noted in Asians, witnessed in high quality subgroup, and apparently noted in intestinal-type gastric cancer.. Our meta-analysis indicates that IL-8 -251 AA genotype is associated with the overall risk of developing gastric cancer and may seem to be more susceptible to overall gastric cancer in Asian populations. IL-8 -251 AA genotype is more associated with the intestinal-type gastric cancer. IL-8 -251 AA genotype is not associated with Helicobacter Pylori infection status in our meta-analysis.. The analyses suggest that IL-8 -251 AA genotype may be an important biomarker of gastric cancer susceptibility for Asians, especially for Chinese Han population, the assumption that needs to be further confirmed in future well-designed studies in China.

Topics: Antigens, Bacterial; Asian People; Bacterial Proteins; China; Gene Frequency; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Odds Ratio; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Risk Factors; Stomach Neoplasms

Genetic epidemiology is an important discipline that is helping to unravel the aetiology and pathogenesis of complex human diseases. In the context of gastrointestinal malignancy, the paradigm model of host genetic influence on disease outcome is H. pylori-associated gastric adenocarcinoma. This cancer represents a classic example of an inflammation-induced malignancy and highlights the importance of host genetics in disease development. This chapter gives an insight into how genetic epidemiology can play an important role in the development of gastric cancer. Increasing our understanding of host genetics in cancer development may allow particularly susceptible individuals to be targeted for screening or treatment to reduce risk of future malignant transformation.

Topics: Adenocarcinoma; Cyclooxygenase 2; Gastroenteritis; Gastrointestinal Neoplasms; Genes, MHC Class II; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Stomach Neoplasms; Tumor Necrosis Factor-alpha

CD74 is a protein whose initial role in antigen presentation was recognized two decades ago. Recent studies have revealed that it has additional functions as a receptor for macrophage migration inhibitory factor and as a receptor for an important human pathogen, Helicobacter pylori (H pylori). The role of CD74 as a receptor is important because after binding of migration inhibitory factor or H pylori, NF-kappaB and Erk1/2 activation occurs, along with the induction of proinflammatory cytokine secretion. This review provides an up-to-date account of the functions of CD74 and how it might be involved in inflammation and cancer within the gastrointestinal tract.

Topics: Antigen Presentation; Antigens, Differentiation, B-Lymphocyte; Gastrointestinal Neoplasms; Helicobacter Infections; Helicobacter pylori; Histocompatibility Antigens Class II; Humans; Inflammation; Interleukin-8; Macrophage Migration-Inhibitory Factors; Protein Isoforms; Signal Transduction

Recently, the role of serine proteinases in the pathogenesis of inflammation and autoimmune diseases via interaction with the proteinase-activated receptor (PAR) has attracted attention. Activation of PAR has a pro-inflammatory effect through the overproduction of inflammatory cytokines such as interleukin (IL)-6 and IL-8. PAR(2) activation in human esophageal epithelial cells by trypsin induces NFkappaB- and AP-1-dependent IL-8 production in association with activation of p38 MAPK and ERK1/2, suggesting that esophageal inflammation may be induced by PAR(2) activation via reflux of trypsin. It has been also proposed that Helicobacter pylori (H. pylori) induces PAR expression in the gastric epithelial cells and H. pylori-derived serine proteinase promotes IL-8 production via PAR in the epithelial cells. In addition, an increase of PAR-dependent IL-8 production has been observed in H. pylori-infected human gastric mucosa, suggesting an important role for PAR(2) in the modulation of gastric inflammation associated with H. pylori. Recent studies have strongly indicated that tryptase and PAR are implicated in the pathogenesis of inflammatory bowel disease and experimental colitis. We demonstrated that anti-tryptase therapy may become a new therapeutic strategy in human ulcerative colitis. Thus, the role of PAR in the gastrointestinal tract has been gradually clarified, but further investigations are needed because the receptor has a variety of functions.

Topics: Animals; Drug Delivery Systems; Gastroenteritis; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Protease Inhibitors; Receptor, PAR-2; Trypsin

Helicobacter pylori infects almost 50% of the world population and is the major cause of gastroduodenal diseases. H. pylori colonizes the gastric mucosa, activates Toll-like and Nod-like receptors, and usually elicits a T helper 1 (Th1) type of immune response, fully polarized in peptic ulcer patients. Among several bacterial factors, the neutrophil-activating protein represents a key factor driving Th1 inflammation. A complex and fascinating balance between H. pylori and host factors takes part in the gastric niche and allows the majority of infected individuals to be without any symptom during their entire life. Novel insights into the innate and adaptive responses against H. pylori, dealing with regulatory T cells and cytokines, CTLA-4 molecule, cholesterol glucosylation, and immune evasion have been elucidated during the past year and are discussed for the development of an effective vaccine.

Topics: Animals; Bacterial Vaccines; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Male; Mice; Th1 Cells; Th2 Cells

Oxygen radicals are supposed to be involved in inflammation and cell proliferation. Helicobacter pylori induces decrease in antioxidant defense factors, such as GSH, mucus and constitutive nitric oxide (NO), gastric mucosal injury and inflammation. Inflammation and injury might be caused by oxidant-mediated expression of inflammatory cytokine interleukin-8 (IL-8) and inflammatory enzymes such as cyclooxtgenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), which were mediated by oxidant-sensitive transcription factors such as nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), possibly with mitogen activated protein kinase (MAPK) activation. H. pylori-induced alterations in protein expression demonstrate the involvement of oxidative stress in the pathogenesis of H. pylori-induced gastric diseases. The differentially expressed genes and proteins may be useful as prognostic indices for gastric diseases associated with H. pylori infection. In conclusion, oxygen radicals are produced in gastric epithelial cells infected with H. pylori, which may reduce the antioxidant defense mechanism and turn on the expression of inflammatory genes, adhesion molecules and mediators stimulating cell proliferation, as well as defensive molecular chaperones in gastric epithelial cells.

Topics: Animals; Cyclooxygenase 2; Gastric Mucosa; Gene Expression Regulation; Helicobacter Infections; Humans; Interleukin-8; Membrane Proteins; Oxidative Stress; Reactive Oxygen Species; Stomach Diseases

Topics: Apoptosis; Cell Proliferation; Epithelial Cells; Gastric Mucosa; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Oxidative Stress

Topics: Bacterial Proteins; Cells, Cultured; Chaperonin 60; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Interleukin-8; Recombinant Proteins; Toll-Like Receptors

Topics: Alleles; Gastritis, Atrophic; Helicobacter Infections; Humans; Interleukin-8; Polymorphism, Genetic; Transcription, Genetic

Topics: Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-8; Neutrophil Infiltration; Stomach Ulcer

Topics: Duodenal Ulcer; Gastric Acid; Gastrins; Gastroesophageal Reflux; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8

Helicobacter pylori(H. pylori) is an important pathogenic factor for gastroduodenal ulcers and gastric cancer. The level of gastric acid output may influence the outcome of those diseases. With low acid output, H. pylori can spread to the corpus of the stomach, resulting in progression to atrophic gastritis. It may cause an increased risk of gastric cancer and ulcer. In contrast, with high output, H. pylori is confined in the gastric antrum, which develops antrum-predominant gastritis. This may contribute to an increased risk of duodenal ulcer. It is well known that inflammatory cytokines including interleukin (IL)-1 beta, IL-8 and tumor necrosis factor alpha modulate gastric acid secretion. Therefore, the host immune response by the cytokines may cause these disparate pathways in gastric acid secretion.

Topics: Gastric Acid; Gastritis, Atrophic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Peptic Ulcer; Tumor Necrosis Factor-alpha

H. pylori colonisation of the stomach causes the recruitment of the inflammatory cells by the adherence of the bacteria with the epithelium and the release of factors of virulence either to the contact (oipA or other soluble factors) or in the cell by translocation (CagA). Such contact triggers interleukin 8 expression in the epithelial cell and attracts lymphocytes and monocytes into the chorion. Bacterial lipopolysaccharide and urease support the activation of these inflammatory cells. The lymphocytes produce pro-inflammatory cytokines, which direct the immune response towards the Th1 pathway. The variability of the inflammatory response depends on hereditary factors of the host such as the interleukin 1 genotypes, which determine the level of the pro-inflammatory cytokine expression, and of bacterial factors such as the cag pathogenicity island, the lipopolysaccharide and the vacuolating toxin, vacA. The mucosal inflammation provokes apoptosis and atrophy of the epithelial cells through the effect of pro-inflammatory cytokines and free radicals. Epithelial proliferation is a consequence of excessive apoptosis caused by the infection. It is stimulated by the expression of inducible cyclo-oxygenase and inducible nitric oxide synthase. The development of atrophic gastritis towards cancer is supported by nitric oxide which has a mutagenic effect on DNA and inhibits p53 protein and by the bacterium itself which decreases DNA mismatch repairing activity. The gastritis induced by Helicobacter pylori changes acid secretion according to the prevalent location of the gastritis in the antrum or in the gastric body. Prevalent gastritis in the gastric body causes hypochlorhydria by reducing the release of histamin from ECL cells and inhibiting the parietal cells through the effect of tumor necrosis factor and interleukin 1-beta. Hypochlorhydria is more marked among patients having a pro-inflammatory genotype for interleukin 1-beta and those infected by bacteria with virulence factors. In the event of antrum predominant gastritis, the pro-inflammatory cytokines cause a reduction of somatostatin and gastrin releases from the D and the G cells, respectively. The result of all is increased maximal acid output and the meal-stimulated acid secretion.

Topics: Apoptosis; Atrophy; Cytokines; DNA Damage; DNA Repair; Gastric Acid; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Lymphocytes

Topics: Animals; Chronic Disease; Gastric Mucosa; Gastritis; Helicobacter Infections; Humans; Interleukin-8; Microcirculation; Neutrophil Activation

Topics: Animals; Autoimmunity; Bacterial Adhesion; Chaperonin 60; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8

Topics: Adhesins, Bacterial; Animals; Bacterial Outer Membrane Proteins; Bacterial Proteins; Carrier Proteins; Evolution, Molecular; Genome, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mutation; Peptic Ulcer

Topics: Animals; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillus; Peptic Ulcer; Probiotics

Gastric inflammation is a highly complex biochemical protective response to cellular/tissue injury. When this process occurs in an uncontrolled manner, the result is excessive cellular/tissue damage that results chronic inflammation and destruction of normal tissue. Current evidence suggests that Helicobacter pylori (H. pylori) infection and nonsteroidal anti-inflammatory drug (NSAID) ingestion are major causative factors in the pathogenesis of gastric mucosal injury in humans. In response to H. pylori infection or NSAID, neutrophils are recruited to the site of inflammation and generate reactive oxygen and nitrogen species and proteases. However, neutrophils are not able to kill the bacteria that live in the gastric mucus, and compounds produced by activated neutrophils themselves may be potentially harmful for normal tissue. It has been shown that leukocyte-vascular endothelial cell interaction is regulated by various cell adhesion molecules, and that this interaction is directly or indirectly modified by many factors, the origin of which is H. pylori and NSAIDs. This review describes the potential role of neutrophils and neutrophil-associated inflammation for gastric oxidative stress and injury induced by H. pylori and/or NSAID.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Adhesion Molecules; Chloramines; Endothelium, Vascular; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Neutrophils

Evidence is accumulating that interleukin-8 (IL-8), discovered as a potent neutrophil chemotactic factor, plays a crucial role in establishing acute neutrophil-mediated inflammation by exerting various activities against non-leukocytic as well as leukocytic cells. Various types of cells can rapidly produce a large amount of IL-8 upon stimulation with inflammatory stimuli, such as lipopolysaccharide, IL-1, and tumor necrosis factor (TNF). However, IL-8 production is tightly regulated at several levels, particularly at the transcriptional levels to prevent aberrant production. Our previous experiments demonstrated that IL-8 gene transcription requires the cooperative activation of a transcription factor, NF-kappa B, with an additional transcription factor, AP-1 or NF-IL6, depending on types of cells and stimuli. In addition, we recently observed that infection with Helicobacter pylori or cytomegalovirus activated NF-kappa B, therapy inducing IL-8 protein secretion as well as IL-8 gene transcription. Moreover, IL-8, in turn, enhanced cytomegalovirus replication and infectious virion production. Collectively, these results suggest the potential involvement of IL-8 in the pathology of bacterial or viral infection.

Topics: Cytomegalovirus; Cytomegalovirus Infections; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Transcription Factors; Transcription, Genetic; Virus Replication

Inflammatory response to Helicobacter pylori is characterized by infiltration of neutrophils, monocytes, and lymphocytes into the gastric mucosa. Interleukin-8 (IL-8), a prototype of the CXC-chemokine subfamily, may be a key modulator in inducing neutrophil migration and activation in H. pylori-infected gastric mucosa. IL-8 is produced by gastric epithelial cells in response to H. pylori infection, and IL-8 expression is induced by local production of proinflammatory cytokines and attachment of H. pylori organisms to the gastric epithelial cell surface. Multiple genes in the H. pylori cag pathogenicity island seem to be involved in inducing the epithelial IL-8 response to H. pylori attachment. Activation of the transcription factor, nuclear factor kappaB (NF-kappaB), is associated with this IL-8 response. Reactive oxygen intermediates whose production is increased in H. pylori-infected gastric mucosa may also modulate IL-8 expression in the gastric mucosa. Recent reports also suggest that the local production of CC-chemokines, another chemokine subfamily, is important in H. pylori-associated gastritis.

Topics: Antigens, Bacterial; Bacterial Proteins; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Reactive Oxygen Species

Infection of Helicobacter pylori causes chronic gastritis and plays an important role in the pathogenesis of gastroduodenal ulceration. H. pylori has also been suggested to be involved in the genesis of adenocarcincoma and MALT lymphoma of the stomach. H. pylori infection is associated with increased gastric epithelial proliferation, which can be reversed by a successful eradication of the organism. Although the mechanisms of increased gastric epithelial proliferation is not known, the enhanced epithelial proliferation is important in developing gastric carcinoma. Whether or not H. pylori de nove stimulates gastric epithelial proliferation is controversial, but gastric infection with H. pylori activates a mucosal inflammatory response by consisting of large numbers of polymorphonuclear and mononuclear cells, that also includes expression of various cytokines including interleukin-8. We review the mechanisms of H. pylori in enhanced gastric epithelial cell proliferation and cytokines in patients with H. pylori infection.

Topics: Animals; Cytokines; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Neutrophils; Peptic Ulcer; Stomach Neoplasms

The gastroduodenal response to chronic Helicobacter pylori infection is characterized by the infiltration of plasma cells, lymphocytes, neutrophils and monocytes into the mucosa. Eradication studies have shown that this inflammatory response represents a specific reaction to the presence of H. pylori. As well as stimulating specific local T and B cell responses and a systemic antibody response, H. pylori infection also induces a local pro-inflammatory cytokine response. Interleukin-8 (IL-8), which is expressed and secreted by gastric epithelial cells, may be an important host mediator inducing neutrophil migration and activation. IL-8 mRNA and protein secretion in gastric epithelial cell lines can be up-regulated by the cytokines tumour necrosis factor-alpha and IL-1 and also by type I strains of H. pylori (expressing the vacuolating toxin and cytotoxin-associated protein, CagA). The gastric epithelium thus plays an active role in mucosal defence. Neutrophil activation and the production of reactive oxygen metabolites will be induced directly by bacterial factors and indirectly via host-derived cytokines, products of complement activation and bioactive lipids. Strain variation in the induction of both IL-8 from epithelial cells and the oxidative burst in neutrophils may be an important factor determining the extent of mucosal injury. There is now increasing evidence from both in vivo and in vitro studies that type I strains induce an enhanced inflammatory response and mucosal damage. An understanding of the bacterial mediators of mucosal inflammation is important in elucidating the role of chronic H. pylori infection in the pathogenesis of gastroduodenal disease.

Topics: Chronic Disease; Cytokines; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Mucosal; Interleukin-8; Neutrophil Activation; Peptic Ulcer

Helicobacter pylori infection is characterized by an inflammatory response in the gastric epithelium, the intensity of which appears to be type-strain specific. Infections caused by Type 1 H. pylori organisms, i.e., those expressing VacA (the cytotoxin) and CagA (the cytotoxin-associated protein), are associated with a strong polymorph mucosal infiltration in vivo, and with increased secretion of interleukin-8 by epithelial cells. The inflammatory potential of Type II strains (non-cytotoxic, VacA- and CagA-negative) is probably less pronounced. The small urease subunit, porins, and other substances produced by H. pylori show neutrophil chemotactic activities in vitro. These bacterial components promote the adhesion of polymorphs to endothelial cells and stimulate polymorphs to generate oxygen reactive metabolites. This can severely damage the gastroduodenal mucosa.

Topics: Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8

To review the role of interleukin (IL)-8 in the immunopathology of Helicobacter pylori-associated gastroduodenal disease.. Literature review.. In H. pylori infection, IL-8 secretion by the gastric mucosa is increased, particularly in patients with active neutrophilic gastritis. Immunoreactive IL-8 is evident in the epithelium of histologically normal gastric mucosa but epithelial IL-8 expression is increased in H. pylori-associated chronic gastritis. Gastric epithelial cell lines constitutively express IL-8 messenger (m)RNA and IL-8 message and protein secretion can be upregulated by the cytokines tumour necrosis factor-alpha, IL-1 alpha and IL-1 beta. H. pylori also directly induces epithelial IL-8 expression in a strain-specific manner. Cytotoxic strains expressing the CagA protein upregulate IL-8 mRNA and IL-8 protein secretion.. IL-8 is an important chemotactic and activating factor for neutrophils. The secretion of IL-8 by epithelial cells is probably a key factor in host defences at mucosal sites, permitting a rapid polymorph response against infectious agents. If defence mechanisms fail and chronic infection results, continued upregulation of IL-8 and neutrophil activation could lead to mucosal damage and increased free radical formation. Mucosal IL-8 production in H. pylori infection may be an important factor in the immunopathogenesis of peptic ulcer disease and also be of relevance to gastric carcinogenesis.

Topics: Duodenal Diseases; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Neutrophil Activation; Stomach Ulcer

Topics: Anti-Inflammatory Agents; Brassica; Cytokines; Gastrins; Gastritis, Atrophic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-18; Interleukin-8; Pepsinogen A; Tumor Necrosis Factor-alpha; Urea

Helicobacter pylori is remarkable for its genetic variation; yet, little is known about its genetic changes during early stages of human infection, as the bacteria adapt to their new environment. We analyzed genome and methylome variations in a fully virulent strain of H pylori during experimental infection.. We performed a randomized Phase I/II, observer-blind, placebo-controlled study of 12 healthy, H pylori-negative adults in Germany from October 2008 through March 2010. The volunteers were given a prophylactic vaccine candidate (n = 7) or placebo (n = 5) and then challenged with H pylori strain BCM-300. Biopsy samples were collected and H pylori were isolated. Genomes of the challenge strain and 12 reisolates, obtained 12 weeks after (or in 1 case, 62 weeks after) infection were sequenced by single-molecule, real-time technology, which, in parallel, permitted determination of genome-wide methylation patterns for all strains. Functional effects of genetic changes observed in H pylori strains during human infection were assessed by measuring release of interleukin 8 from AGS cells (to detect cag pathogenicity island function), neutral red uptake (to detect vacuolating cytotoxin activity), and adhesion assays.. The observed mutation rate was in agreement with rates previously determined from patients with chronic H pylori infections, without evidence of a mutation burst. A loss of cag pathogenicity island function was observed in 3 reisolates. In addition, 3 reisolates from the vaccine group acquired mutations in the vacuolating cytotoxin gene vacA, resulting in loss of vacuolization activity. We observed interstrain variation in methylomes due to phase variation in genes encoding methyltransferases.. We analyzed adaptation of a fully virulent strain of H pylori to 12 different volunteers to obtain a robust estimate of the frequency of genetic and epigenetic changes in the absence of interstrain recombination. Our findings indicate that the large amount of genetic variation in H pylori poses a challenge to vaccine development. ClinicalTrials.gov no: NCT00736476.

Topics: Antigens, Bacterial; Bacterial Adhesion; Bacterial Proteins; Bacterial Vaccines; Biopsy; DNA Methylation; Epigenesis, Genetic; Gene Expression Regulation, Bacterial; Genome, Bacterial; Genomic Islands; Genotype; Germany; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Mutation; Phenotype; Polymorphism, Single Nucleotide; Time Factors; Virulence

Helicobacter pylori infection of the lining of the stomach induces an array of inflammatory cytokine production leading to gastritis and peptic ulcer disease. The aim of this study was to investigate the effect of curcumin on the production of interleukin (IL)-8, IL-1beta, tumor necrosis factor (TNF)-alpha and cyclooxygenase (COX)-2 in gastric mucosa from H. pylori-infected gastritis patients. Patients were randomly assigned to receive either OAM (Omeprazole, Amoxicillin and Metronidazole) treatment or a course of curcumin. Gastric biopsies were collected before and after treatment and were examined for the level of inflammatory cytokines mRNA by semi-quantitative reverse transcription polymerase chain reaction. The eradication rate of H. pylori in patients that received OAM treatment was significantly higher than the patients that received curcumin (78.9% versus 5.9%). The levels of IL-8 mRNA expression in the OAM group significantly decreased after treatment, but no changes of other cytokines were found. This emphasizes an important role of IL-8 in H. pylori infection. The decreases of cytokine production were not found in the curcumin group. We concluded that curcumin alone may have limited anti-bactericidal effect on H. pylori, and on the production of inflammatory cytokines. Nevertheless, other studies have reported that patients treated with curcumin had relieved symptoms. Further investigation should be carried out as the use of curcumin in combination with therapeutic regimens may be beneficial as an alternative treatment.

Topics: Adult; Amoxicillin; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Bacterial Load; Curcumin; Drug Therapy, Combination; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Metronidazole; Middle Aged; Omeprazole

Helicobacter pylori (H. pylori) is associated with chronic gastritis and gastric carcinogenesis. The effects of nonsteroidal anti-inflammatory drugs (NSAIDs), which exert chemopreventive effects on several cancers, on H. pylori-induced gastritis remain unknown. We investigated the effects of NSAIDs on gastric inflammation and the kinetics of gastric epithelial cells in H. pylori-induced gastritis.. Patients with rheumatoid arthritis or osteoarthritis who took NSAIDs for more than 1 month and complained of dyspeptic symptoms were recruited for this study. Patients not on any NSAIDs were included as non-NSAID user controls. All patients underwent diagnostic testing for H. pylori infection, esophagogastroduodenoscopy, and gastric biopsies. Neutrophil infiltration into gastric mucosa, expression of inducible nitric oxide synthase (iNOS), and apoptosis and proliferation of gastric epithelial cells were evaluated by immunohistochemistry. In an in vitro study, the effects of NSAIDs on production of interleukin (IL)-8 induced by H. pylori in a gastric epithelial cell line (AGS) were determined.. Numbers of neutrophils infiltrating the gastric mucosa, iNOS-expressing inflammatory cells and apoptotic cells, and proliferating cells in gastric epithelium were higher in H. pylori-positive groups than H. pylori-negative groups. Among H. pyloripositive groups, these parameters were lower in NSAID users than in non-NSAID users. NSAIDs inhibited the production of IL-8 induced by H. pylori in AGS cells.. These findings suggest that long-term use of NSAIDs normalizes the kinetics of gastric epithelial cells in patients with H. pylori infection by attenuating gastric mucosal inflammation, which may result in prevention of the gastric carcinogenesis associated with H. pylori infection.

Topics: Adult; Aged; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Cell Line, Tumor; Epithelial Cells; Female; Gastric Mucosa; Gene Expression Regulation, Enzymologic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Infiltration; Nitric Oxide Synthase Type II; Osteoarthritis; Time Factors

Even with the most effective treatment, Helicobacter pylori eradication is difficult in some patients. Therefore, patients sometimes require acid-suppressive therapy without H. pylori eradication. It has been reported that ranitidine inhibits neutrophil activation, whereas famotidine does not. However, few studies have been published concerning the activation of neutrophils before and after treatment using clinical doses of histamine-2 receptor antagonists in patients with H. pylori infection.. To examine the effects of neutrophil activation after treatment with three different histamine-2 receptor antagonists.. This prospective, open-label, randomised, parallel-group study was conducted. Thirty patients with H. pylori infection were enrolled. These subjects were randomly assigned to receive one of the following treatments: (a) 150 mg ranitidine, (b) 20mg famotidine, or (c) 10 mg lafutidine b.d., for 4 weeks. Before and after histamine-2 receptor antagonist treatment, histological findings, myeloperoxidase activity, and interleukin-8 in the gastric mucosa were evaluated.. On the basis of the histological findings between before and after histamine-2 receptor antagonist treatment, no significant differences were found in any groups. Similarly, there were no significant differences in myeloperoxidase activity or interleukin-8 levels.. In patients with H. pylori, when used at clinical doses, any histamine-2 receptor antagonists can be used without concerning about inhibition of neutrophil activation.

Topics: Acetamides; Adult; Dyspepsia; Famotidine; Female; Helicobacter Infections; Helicobacter pylori; Histamine H2 Antagonists; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Activation; Peroxidase; Piperidines; Pyridines; Ranitidine

Helicobacter pylori infection is an important risk factor for gastric diseases. Some probiotics are useful for suppressing H. pylori infection. Bifidobacterium bifidum YIT 4007 can improve the experimental gastric injury in rats and the disease stages on the gastric mucosa in peptic ulcer patients. We evaluated the fermented milk using a clone (BF-1) having the stronger ability to survive in the product than this parent strain to clarify the in vitro suppressive effect of BF-1 on H. pylori and the in vivo efficacy of BF-1 fermented milk on H. pylori and gastric health. In the mixed culture assay of BF-1 and H. pylori, the number of pathogens was decreased such that it was not detected after 48 h in the Brucella broth with a decrease in pH values. In the cell culture experiment with human gastric cells, the H. pylori infection-induced IL-8 secretion was suppressed by the preincubation of BF-1. In a human study of 12-wk ingestion (BF-1 group, n = 40; placebo group, n = 39) with a randomized double-blind placebo-control design, the H. pylori urease activity and gastric situation were evaluated using a urea breath test (UBT) and the serum pepsinogen (PG) levels as biomarkers for inflammation or atrophy, respectively. In the H. pylori-positive subjects, the difference (DeltaUBT) of the UBT value from the baseline value in the BF-1 group (n = 34) was lower than that in the placebo group (n = 35) at 8 wk. The baseline UBT values showed a negative correlation with DeltaUBT values at 8 and 12 wk in the BF-1 group but not in the placebo. In the PG-positive subjects classified by the PG test method, the BF-1 group was lower in DeltaUBT values than the placebo group at 8 and 12 wk. In the active gastritis class by PG levels, the BF-1 group was lower in their DeltaUBT values than the placebo at 8 and 12 wk. The PG I levels in the BF-1 group were lower than the placebo at 12 wk. The PG II levels in the BF-1 group did not change during the ingestion period, but the placebo was increased. The PG I/II ratios slightly decreased from baseline at 12 and 20 wk in the BF-1 and placebo groups. These patterns were also observed in the H. pylori-positive subjects. The improving rates of upper gastrointestinal symptomatic subjects and total symptom numbers in the BF-1 group were higher than those in the placebo. These results indicate that BF-1 fermented milk may affect H. pylori infection or its activity, gastric mucosal situation, and the emergence of upper gastrointestinal symp

Topics: Animals; Bifidobacterium; Biomarkers; Breath Tests; Cell Line; Double-Blind Method; Female; Fermentation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Milk; Pepsinogen A; Probiotics; Treatment Outcome; Urease

The role of teprenone in Helicobacter pylori-associated gastritis has yet to be determined. To investigate the effect of teprenone on inflammatory cell infiltration, and on H. pylori colonization of the gastric mucosa in H. pylori-infected patients, we first compared the effect of teprenone with that of both histamine H2 receptor antagonists (H2-RA) and sucralfate on the histological scores of H. pylori gastritis. We then examined its in vitro effect on H. pylori-induced interleukin (IL)-8 production in MKN28 gastric epithelial cells.. A total of 68 patients were divided into three groups, each group undergoing a 3-month treatment with either teprenone (150 mg/day), H2-RA (nizatidine, 300 mg/day), or sucralfate (3 g/day). All subjects underwent endoscopic examination of the stomach before and after treatment. IL-8 production in MKN28 gastric epithelial cells was measured by enzyme-linked immunosorbent assay (ELISA).. Following treatment, the teprenone group showed a significant decrease in both neutrophil infiltration and H. pylori density of the corpus (before vs. after: 2.49 +/- 0.22 vs. 2.15 +/- 0.23, p =.009; 2.36 +/- 0.25 vs. 2.00 +/- 0.24, p =.035, respectively), with no significant differences seen in either the sucralfate or H2-RA groups. Teprenone inhibited H. pylori-enhanced IL-8 production in MKN28 gastric epithelial cells in vitro, in a dose-dependent manner.. Teprenone may modify corpus H. pylori-associated gastritis through its effect on neutrophil infiltration and H. pylori density, in part by its inhibition of IL-8 production in the gastric mucosa.

Topics: Anti-Ulcer Agents; Biopsy; Cell Line; Diterpenes; Epithelial Cells; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Histamine H2 Antagonists; Humans; Interleukin-8; Male; Middle Aged; Neutrophil Infiltration; Nizatidine; Pepsinogen A; Pepsinogen C; Sucralfate; Urease

Inducible nitric oxide synthase (iNOS) and interleukin 8 (IL-8) are positive in approximately 50% of Helicobacter pylori-related diseases but it is not clear whether oxidative stress is also present in H. pylori asymptomatic humans. Our aim was to study the expression of iNOS, superoxide dismutase, catalase and IL-8 production in H. pylori-infected asymptomatic humans, and to investigate the effect of eradication of H. pylori.. Biopsies of corpus and antrum of asymptomatic H. pylori positive and negative humans served for determination of the gastritis score and H. pylori status; iNOS was measured by reverse transcriptase polymerase chain reaction and immunohistochemistry and superoxide dismutase and catalase by immunohistochemistry. IL-8 in biopsies was assessed by enzyme-linked immunosorbent assay.. Immunostaining of iNOS, catalase and superoxide dismutase was significantly associated with H. pylori infection and was localized to inflammatory cells. IL-8 concentrations were greater in the H. pylori positive than H. pylori negative group and decreased after bacterial eradication. A decrease in staining for iNOS and catalase was observed after H. pylori eradication.. INOS and antioxidant enzymes are present in gastric biopsies of asymptomatic H. pylori positive humans. Eradication caused a significant decrease in staining for iNOS and catalase. These results indicate that oxidative stress occurs in asymptomatic patients and can be modulated by H. pylori eradication.

Topics: Adult; Amoxicillin; Anti-Bacterial Agents; Anti-Ulcer Agents; Biomarkers; Biopsy; Catalase; Clarithromycin; Drug Therapy, Combination; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Omeprazole; Oxidative Stress; Superoxide Dismutase

Production of cytokines along with increased activity of nitric oxide synthase has been implicated as one of the contributing mechanisms of Helicobacter pylori-mediated gastroduodenal diseases. We aimed to evaluate the effect of rebamipide in treating Helicobacter pylori-associated duodenal ulcers in terms of cytokine production and nitrosative damage of the gastric mucosa. In patients with duodenal ulcers, rebamipide or placebo were given randomly after eradication. Mucosal cytokine production was measured by enzyme linked immunoassay, and nitrotyrosine immunoexpression was measured by immunohistochemistry. The inflammatory activity and degree of neutrophil infiltration were graded accordingly. The mucosal production of RANTES, interleukin-8, and TNF-alpha showed a significant decrease after eradication in patients with rebamipide after-treatment. The nitrotyrosine immunoreactivity of gastric epithelium was significantly decreased in the rebamipide group. Rebamipide treatment after eradication resulted in a significant reduction in chemokine production along with nitrotyrosine immunoexpression in Helicobacter pylori-associated duodenal ulcers.

Topics: Adult; Alanine; Antioxidants; Chemokine CCL5; Cytokines; Drug Therapy, Combination; Duodenal Ulcer; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Male; Quinolones; Tumor Necrosis Factor-alpha; Tyrosine

There is accumulating evidence for the role of Helicobacter pylori in the development of gastric cancer as well as of lymphomas that arise in mucosa-associated lymphoid tissue (MALT). We reported recently that gastric cancer patients show high prevalence of cagA-positive H. pylori and express gastrin and gastrin receptors enabling them to stimulate tumour growth in autocrine fashion.. Since the H. pylori infection is considered to be more strongly associated with MALT lymphoma than with gastric cancer, we decided to determine the gastrin and its receptors' mRNA expression and gastrin content in this tumour as well as the release of this hormone both into plasma and gastric lumen. Twenty MALT lymphoma patients were compared with 100 age- and gender-matched controls with similar dyspeptic symptoms.. The overall H. pylori seropositivity in MALT lymphoma was about 90% and CagA positivity was 70%, compared to 56% and 33%, respectively, in controls. The serum gastrin in MALT lymphoma was about sixfold higher than in controls while gastric luminal gastrin in these patients was over 70 times higher than in controls. Gastrin content in tumour was about 10-fold higher than in antral mucosa. Gastrin and gastrin-receptor (CCKB-receptor) mRNA were detected by reverse transcriptase-polymerase chain reaction in cancer tissue whilst in the fundic and antral mucosa, only enhanced expression of CCKB-receptor mRNA and gastrin mRNA was detected, respectively. Histamine stimulation in MALT lymphoma induced acid secretion that was only about 30% of control value due to atrophic gastritis. This study confirms an important role of CagA-positive H. pylori in the pathogenesis of MALT lymphoma and shows that this lymphoma is capable of synthesizing and releasing potent growth promoting gastrin, possibly due to the action on G-cells of H. pylori-originated Nalpha-methyl histamine and cytokines (tumour necrosis factor alpha and interleukin-8).. Gastric MALT lymphoma is closely linked to CagA-positive H. pylori infection. Gastrin and its receptors may be implicated in the pathogenesis of gastric lymphoma.

Topics: Adult; Aged; Antigens, Bacterial; Bacterial Proteins; Cytokines; Female; Gastric Acid; Gastric Mucosa; Gastrins; Helicobacter Infections; Helicobacter pylori; Histamine; Humans; Interleukin-8; Lymphoma, B-Cell, Marginal Zone; Male; Middle Aged; Radioimmunoassay; Receptors, Cholecystokinin; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms

The purpose of the present study was to examine the association between interleukin-8 (IL-8) in the gastric body due to Helicobacter pylori infection and histological gastritis, as well as elucidating the effect of acid secretion inhibitors on H. pylori associated body gastritis in duodenal ulcer patients.. Twenty H. pylori-negative patients, 20 H. pylori-positive patients with chronic gastritis without peptic ulceration, and 20 H. pylori-positive duodenal ulcer patients (DU) were studied. Four biopsy samples were taken, each from the greater curvature of the antrum and body of the stomach. Biopsies were histologically investigated by ELISA to determine the density of H. pylori, the degree of neutrophil infiltration and the IL-8 concentration in the mucosa.. In the gastric mucosa of H. pylori-negative subjects, no IL-8 and hardly any neutrophil infiltration were observed. In contrast, enhanced IL-8 production and increased neutrophil infiltration were present in those infected with H. pylori. In H. pylori-positive patients, a significant correlation was observed between the IL-8 concentration and the degree of neutrophil infiltration, but no correlation was found in the body mucosa of those with DU. Twelve of 20 DU patients demonstrated hardly any neutrophil infiltration, despite the increased mucosal IL-8 content in the body. The administration of omeprazole in DU patients markedly increased mucosal neutrophil infiltration even though it did not cause any significant change in the H. pylori density and IL-8 concentration in the body. Although the effect of omeprazole was transient, a significant increase in neutrophil infiltration continued in comparison with the status before omeprazole administration in those subsequently undergoing maintenance treatment with H2-blockers.. In H. pylori-positive chronic gastritis, IL-8 concentration is enhanced in the mucosa of the body, and is associated with increased neutrophil infiltration. However, in DU patients, despite increases in body IL-8 concentration, neutrophil infiltration is reduced and the gastritis may be localized in the antrum.

Topics: Adult; Aged; Anti-Ulcer Agents; Duodenal Ulcer; Enzyme Inhibitors; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Omeprazole; Proton Pump Inhibitors; Stomach

Helicobacter pylori (Hp) is one of the most prevalent pathogenic microorganisms in the world, which is related to gastric ulcer.. To observe the effect of lansoprazole and omeprazole combined with antibiotics on gastric juice pH and inflammatory factors in elderly patients with Hp positive gastric ulcer.. This study was a prospective observation study.. This study was performed in Department of Gastroenterology, First Affiliated Hospital of Soochow University.. One hundred and ten elder patients with Hp positive gastric ulcer admitted to our hospital from January 2019 to May 2020.. The control group was treated with omeprazole combined with antibiotics, and the observation group was treated with lansoprazole combined with antibiotics.. The level of gastric juice pH, interleukin-1 (IL-1), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) and heat shock protein-70 (HSP-70).. The changes of gastric juice pH value, IL-1, IL-8, TNF-α and HSP-70 levels before and after treatment were detected in the two groups. The total effective rate, Hp eradication rate, mature type of regenerated mucosal tissue surrounding ulcer and adverse reaction rate were statistically analyzed.. The total effective rate and Hp eradication rate in the observation group were higher than those in the control group, while the adverse reaction rate in the observation group was lower than that in the control group (P < .05). After treatment, the pH value of gastric juice and HSP-70 in the observation group were higher than those in the control group, while the IL-1, IL-8 and TNF-α were lower than those in the control group (P < .05). The mature type of regenerated mucosal tissue structure around ulcer in the observation group was better than that in the control group (P < .05).. The overall effect of lansoprazole combined with antibiotics in the treatment of Hp positive gastric ulcer in the elderly is better than that of omeprazole combined with antibiotics.

Topics: Aged; Anti-Bacterial Agents; Anti-Infective Agents; Anti-Ulcer Agents; Drug Therapy, Combination; Gastric Juice; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Interleukin-1; Interleukin-8; Lansoprazole; Omeprazole; Prospective Studies; Stomach Ulcer; Tumor Necrosis Factor-alpha; Ulcer

The role of inflammatory cytokines, such as tumor necrosis-α (TNF-α) and IL-8, in gastric carcinogenesis has been investigated, but their impact remains to be further elucidated.. In this study, we measured the serum concentrations of these cytokines and H. pylori serostatus in dyspeptic patients, presenting with normal mucosa (NM = 53), chronic gastritis (CG = 94), and gastric cancer (GC = 82), by ELISA.. Moderate levels of TNF-α were detected in the NM group (19.9 ± 19.5 pg/ml), which were nearly doubled in patients with CG (35.7 ± 28.0 pg/ml) and drastically declined in GC patients (1.8 ± 5.9 pg/ml). The serum levels of IL-8, however, were not statistically different amongst these three groups.. TNF-α serum concentration seemed to undergo up- and downregulation, when moving from NM to CG and from CG to GC, respectively. If confirmed in a prospective study, this cytokine can behave as a serum indicator of gastric inflammation and malignant transformation.

Topics: Cytokines; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Prospective Studies; Stomach Neoplasms; Tumor Necrosis Factor-alpha

Helicobacter pylori infection is considered as the major risk factor for gastric adenocarcinoma. Today, the increasing emergence of antibiotic-resistant strains has drastically decreased the eradication rate of H. pylori infection. This study was aimed to investigate the inhibitory and modulatory effects of live and pasteurized Lactobacillus crispatus strain RIGLD-1 on H. pylori adhesion, invasion, and inflammatory response in AGS cell line.. The probiotic potential and properties of L. crispatus were evaluated using several functional and safety tests. Cell viability of AGS cells exposed to varying concentrations of live and pasteurized L. crispatus was assessed by MTT assay. The adhesion and invasion abilities of H. pylori exposed to either live or pasteurized L. crispatus were examined by gentamycin protection assay. The mRNA expression of IL-1β, IL-6, IL-8, TNF-α, IL-10, and TGF-ß genes was determined by RT-qPCR from coinfected AGS cells. ELISA was used for the detection of IL-8 secretion from treated cells. Both live and pasteurized L. crispatus significantly decreased H. pylori adhesion/invasion to AGS cells. In addition, both live and pasteurized L. crispatus modulated H. pylori-induced inflammation by downregulating the mRNA expression of IL-1β, IL-6, IL-8, and TNF-α and upregulating the expression of IL-10, and TGF-ß cytokines in AGS cells. Furthermore, H. pylori-induced IL-8 production was dramatically decreased after treatment with live and pasteurized L. crispatus.. In conclusion, our findings demonstrated that live and pasteurized L. crispatus strain RIGLD-1 are safe, and could be suggested as a potential probiotic candidate against H. pylori colonization and inflammation.

Topics: Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-10; Interleukin-6; Interleukin-8; Lactobacillus crispatus; RNA, Messenger; Tumor Necrosis Factor-alpha

Topics: Cell Line, Tumor; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Kelch-Like ECH-Associated Protein 1; Mitochondria; NF-E2-Related Factor 2; Panax; Plant Extracts; Reactive Oxygen Species

Sumac (Rhus coriaria L.) is a spice and medicinal herb traditionally used in the Mediterranean region and the Middle East. Since we previously demonstrated Sumac biological activity in a model of tumor necrosis factor alpha (TNF-α)-induced skin inflammation, the present work is aimed at further demonstrating a potential role in inflammatory disorders, focusing on gastritis. For this purpose, different polar extracts (water-W, ethanol-water-EW, ethanol-E, ethanol macerated-Em, acetone-Ac, ethylacetate-EtA) were investigated in gastric epithelial cells (GES-1) challenged by TNF-α or H. pylori infection. The ethanolic extracts (E, EW, Em) showed the major phenolic contents, correlating with lower half maximal inhibitory concentrations (IC50s) on the release of interleukin-8 (IL-8, <15 μg/mL) and interleukin-6 (IL-6, <20 μg/mL) induced by TNF-α. Similarly, they inhibited IL-8 release (IC50s < 70 μg/mL) during Helicobacter pylori (H. pylori) infection and exhibited a direct antibacterial activity at comparable concentrations (minimum inhibitory concentration (MIC) = 100 μg/mL). The phenolic content and the bioactivity of EW were maintained after simulated gastric digestion and were associated with nuclear factor kappa B (NF-κB) impairment, considered the main putative anti-inflammatory mechanism. On the contrary, an anti-urease activity was excluded. To the best of our knowledge, this is the first demonstration of the potential role of Sumac as a nutraceutical useful in H. pylori-related gastritis.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents; Dietary Supplements; Epithelial Cells; Ethanol; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Interleukin-6; Interleukin-8; Phenols; Plant Extracts; Rhus; Tumor Necrosis Factor-alpha; Water

Previous reports have shown that

Topics: Animals; Cytotoxins; Exotoxins; Helicobacter Infections; Helicobacter pylori; Interleukin-6; Interleukin-8; Lung Injury; Mice; NF-kappa B; Tumor Necrosis Factor-alpha

Helicobacter bilis, an enterohepatic Helicobacter species, represents a carcinogenic risk factor for cholangiocytes owing to the prevalence of infections in patients with biliary tract cancer, cholecystitis, and pancreaticobiliary maljunction. However, the effect of H. bilis infection on cholangiocytes and the process and mechanism of carcinogenesis are not known. We aimed to determine the effects of H. bilis on cholangiocytes, focusing on inflammation and oxidative stress.. Helicobacter bilis and MMNK-1 cells were cocultured for 24 h and inflammatory cytokine secretion was evaluated. Furthermore, MMNK-1 cell proliferation, intracellular reactive oxidant species (ROS) production, and DNA damage caused by ROS were investigated. All factors were compared with and without H. bilis infection.. Interleukin (IL)-6 and IL-8 secretion were significantly increased in MMNK-1 cocultures with H. bilis (IL-6, 24.3 ± 12.2 vs. 271.1 ± 286.4 pg/ml; IL-8, 167.6 ± 78.7 vs. 1085.1 ± 1047.1 pg/ml, p < .05). MMNK-1 proliferation was also significantly higher in H. bilis cocultures (1.05 ± 0.02 vs. 1.00-fold, respectively; p < .05). Coculturing enhanced the production of ROS in MMNK-1 cells depending on the cell concentration of H. bilis (1.0 vs. 1.17 ± 0.06, p < .05); however, DNA injury was not observed in cocultures with H. bilis (5.35 ± 0.87 vs. 6.08 ± 0.55 pg/μl, p = .06).. Helicobacter bilis infection induced ROS production in and enhanced the proliferation of cholangiocytes.

Topics: Cell Proliferation; Helicobacter; Helicobacter Infections; Humans; Interleukin-6; Interleukin-8; Oxidative Stress; Reactive Oxygen Species

The outer membrane vesicles (OMVs) secreted by Helicobacter pylori contain various bacterial components, such as proteins, phospholipids, toxins, and nucleic acids, including small noncoding RNA (sncRNA), which have regulatory functions in cell envelope structure, metabolism, bacterial communication, biofilm formation, and virulence. We previously showed that knocking out sncRNAs sR-989262 and sR-2509025 at the cellular level increased interleukin 8 (IL-8) levels in mice exposed to OMVs. In this study, we show that immunization with ΔsR-989262 and ΔsR-2509025 OMVs intragastrically significantly increased immunoglobulin G (IgG) and secreted IgA levels in mice compared to wild-type OMVs and without weight changes, which indicated that sncRNA-deficient OMVs are relatively safe to immunize mice. The detection of IgG subtypes IgG1 and IgG2c showed that the sncRNA-deficient OMVs primarily stimulate the T helper 2 (Th2)-mediated immune response. Moreover, levels of the cytokines IL-4, IL-13, gamma interferon (IFN-γ), IL-12 (p40), IL-8, and IL-17 indicate that ΔsR-989262 and ΔsR-2509025 OMVs trigger the Th2-type immune response but primarily trigger a Th1-mediated and Th17-mediated immune response. These findings show that OMV-encapsulated sncRNA plays an important role in regulating the immune response in hosts infected by H. pylori at the animal level. Moreover, they show that knocking out of sR-989262 and sR-2509025 improves the immunogenicity and protective efficacy of OMVs, and this may be beneficial to the design of OMV-based H. pylori vaccines.

Topics: Animals; Bacterial Outer Membrane Proteins; Disease Models, Animal; Helicobacter Infections; Helicobacter pylori; Immunoglobulin G; Interleukin-8; Mice; RNA, Small Untranslated

Here, we aimed to evaluate and compare the anti-Helicobacter pylori activity of potential probiotic Lactiplantibacillus pentosus SLC13 to Lactobacillus gasseri BCRC 14619 T and Lacticaseibacillus rhamnosus LGG. Phenotypic assays including growth curve, cell adhesion, and cellular cytotoxicity were performed to characterize SLC13. Anti-H. pylori activity of lactobacilli was determined by the disk diffusion method and co-culture assay. Exopolysaccharide (EPS) was extracted from lactobacilli to test its immune modulation activity, and IL-8 expression in AGS and GES-1 was determined by RT-qPCR.. All three lactobacilli strains were tolerant to the simulated gastrointestinal conditions. SLC13 showed the highest adhesion ability to AGS and GES-1 cells, compared to LGG and BCRC 14619 T. The coculture assays of SLC13, LGG, and BCRC 14619 T with cells for 4 h showed no significant cytotoxic effects on cells. All tested strains exhibited an inhibitory effect against H. pylori J99. The cell-free supernatant (CFS) of three strains showed activity to inhibit H. pylori urease activity in a dose-dependent manner and the CFS of SLC13 had the highest urease inhibitory activity, compared to LGG and BCRC 14619 T. Only the treatment of AGS cells with SLC13 EPS significantly decreased the IL-8 expression induced by H. pylori infection as compared to cells treated with LGG and BCRC 14619 T EPS.. SLC13 possesses potent antimicrobial activity against H. pylori growth, infection, and H. pylori-induced inflammation. These results suggest that SLC13 and its derivatives have the potential as alternative agents against H. pylori infection and alleviate inflammatory response.

Topics: Bacterial Adhesion; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillus; Probiotics; Urease

Topics: Adenocarcinoma; Animals; Anti-Bacterial Agents; Cell Line, Tumor; Citrus paradisi; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillus plantarum; Laurates; Male; Mice; Mice, Inbred BALB C; Monoglycerides; Plant Extracts; Probiotics; Specific Pathogen-Free Organisms; Stomach; Stomach Neoplasms

Gastric nitric oxide (NO) production in response to. Esophagogastroduodenoscopy was done in 96 dyspepsia patients. Luminal [NO] was measured by chemiluminescence. Biopsies were taken from gastric antrum and corpus for culture and histopathology.. Of the parameters tested, increased gastric [NO] and circulating IL-8 align most consistently and selectively in

Topics: Gases; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Nitric Oxide

Bacteria-derived outer membrane vesicles (OMVs) are commonly associated with various biological activities and functions. Helicobacter pylori-derived OMVs are thought to contribute to pathogenesis. This study aimed to investigate the effects of H. pylori-derived OMVs.. H. pylori strains were isolated from patients with gastritis, gastric ulcer, or gastric cancer using endoscopic biopsy. The U-937, AGS, and MKN-45 cell lines were exposed to H. pylori and H. pylori-derived OMVs. The expression of interleukin 8 (IL-8) messenger RNA (mRNA) was assessed using reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR, and IL-8 secretion was analyzed using enzyme-linked immunosorbent assay. Nuclear factor kappa B (NF-κB) activation was evaluated by Western blotting.. H. pylori and H. pylori-derived OMVs induced the expression of IL-8 mRNA and protein. Importantly, the bacteria induced higher IL-8 mRNA and protein expression than the OMVs. IL-8 expression was induced to different levels in response to H. pylori-derived OMVs from hosts with different gastric diseases. Western blotting revealed the increased phosphorylation and reduced degradation of inhibitor of NF-κB alpha in cells exposed to OMVs.. H. pylori-derived OMVs may aid the development of various gastric diseases by inducing IL-8 production and NF-κB activation.

Topics: Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B

Antibiotics are known to be effective in treating bacterial infectious disease. Changes in microflora and mucosal dysbiosis may take place after antibiotic treatment. We investigated in this research the effect of anti-Helicobacter pylori treatment (AHT) on local immunity of the female genital tract.. The study identified the levels of cytokines IL-8 and TNF-α in vaginal secretion in a group of female patients with Helicobacter-associated acid-related diseases who were or were not treated with antibiotics against Helicobacter Pylori.. Research outcomes turned out that the secretory cytokine (chemokine) IL-8 is dramatically increased in the vaginal mucosa in patients treated with antibiotics, specifically in post-menopause women.. Helicobacter pylori eradication treatment affects the immune status of the female genital tract.

Topics: Female; Helicobacter Infections; Humans; Interleukin-8; Vagina

Topics: Antigens, Bacterial; Cell Nucleus; Cytokines; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; MAP Kinase Signaling System; Microbial Sensitivity Tests; NF-kappa B; NF-kappa B p50 Subunit; Quinazolines; Signal Transduction; Subcellular Fractions; Type IV Secretion Systems

Stomach pathologies develop in a complex interaction between the host's genetic background and H. pylori virulent genes. Thus, our study aimed to compare active chronic gastritis (ACG), and intestinal metaplasia (IM) with inactive chronic gastritis (ICG), according to interleukin polymorphisms of IL6-174 G/C, IL8-251 A/T, IL1β-511 C/T, and IL1RN VNTR taking into account patient gender and H. pylori genotypes. Interleukin polymorphisms were determined by RFLP-PCR and H. pylori genotype by PCR. IL6-174 GC and IL8-251 T allele showed a protective effect in women against ACG development and, conversely, IL8-251 polymorphism showed a risk for men. More virulent H. pylori strains were associated with the IL8-251 T allele and IL1β-511 T allele in the AGC, and the vacA m1 allele and cagE gene from H. pylori was associated with the IM. Analysis of the progression of gastric lesions must take into account host variability genetic associated with genes H. pylori due to the relation between the virulent H. pylori genes and more severe gastric lesions, besides the relevance to the gender to IL6-174 and IL8-251 polymorphisms.

Topics: Female; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-6; Interleukin-8; Interleukins; Male; Polymorphism, Genetic; Stomach Neoplasms

Helicobacter pylori (H. pylori) induces an intense inflammatory response, mediated by proinflammatory cytokines, including interleukin (IL)-6 and its membrane receptor (IL-6R), which activates important signaling pathways in the development of gastric disease and cancer. We investigated the gene and protein expression of IL-6 and IL-6R and the influence of polymorphisms rs1800795, rs1800796, and rs1800797 on its gene expression together with H. pylori infection. Furthermore, an in-silico analysis was performed to support our results. Gastric biopsies were obtained from patients with gastric symptoms and patients with gastric cancer (GC) and were divided into groups (Control, Gastritis, and Cancer). H. pylori was detected by PCR. Real-time-qPCR was employed to determine gene expression, and western blot assay was used to analyze protein expression levels. PCR-RFLP was used to characterize IL-6 polymorphisms. Bioinformatics analyses were performed using the Gene Expression Omnibus (GEO) database and GEO2R to screen out differentially expressed genes (DEGs). H. pylori was detected in 43.3% of the samples. Statistically significant differences were found for IL-6 (P=0.0001) and IL-6R (P=0.0005) genes among the three groups, regardless of the presence of H. pylori. Among patients with H. pylori infection, the IL-6 and IL-6R gene and protein expressions were significantly increased, highlighting IL-6 gene overexpression in patients with GC. No statistically significant differences were found for the rs1800795, rs1800796, and rs1800797 polymorphisms compared to IL-6 gene expression. The results indicated that the IL-6 polymorphisms do not influence its expression, but IL-6 and IL-6R expression seems to be altered by the presence of H. pylori.

Topics: Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-6; Interleukin-8; Stomach Neoplasms

Tumour-derived CXCL8 facilitates the movement of myeloid-derived suppressor cells, which are able to restrain antitumour immune responses to the tumour microenvironment. Kruppel-like factor 4 (KLF4) is a potential tumour suppressor in gastric cancer (GC). However, knowledge regarding correlations between KLF4 and CXCL8 in GC is limited. We use cellular and molecular biological methods to assess whether these two factors interact in GC. Expression CXCL8 and KLF4 was altered in human GC tissues compared to normal gastric tissues in opposite ways. Additionally, cytotoxin-associated gene A protein (CagA) gene transduction or Helicobacter pylori (H. pylori) infection upregulated CXCL8 expression. Knockdown of KLF4 expression increased CXCL8 protein and RNA expression, whereas its overexpression had the opposite effect. CXCL8-mediated enhancement of GC cell migration and proliferation was reversed by upregulation of KLF4 expression. Further mechanistic research revealed that KLF4 binds the CXCL8 promoter, suppressing CXCL8 transcription. Moreover, CXCL8 stimulation reduced KLF4 protein expression and promoted GC cell proliferation and migration, eventually promoting neoplasm growth in vivo. Together, our findings demonstrate that CagA promotes CXCL8 and inhibits KLF4. CXCL8 is a decisive downstream target gene of KLF4, and KLF4 negatively regulates CXCL8 in GC. Furthermore, CXCL8's negative regulation of KLF4 in vivo and in vitro, indicates that CagA may downregulate KLF4 by inducing CXCL8 expression, low expression of KLF4 further promotes that of CXCL8, forming a vicious circle in GC. Targeted KLF4 activation might improve the immunosuppressive microenvironment through direct negative regulation of CXCL8, providing a new potential target to strengthen the efficacy of immunotherapy in GC patients.

Topics: Cell Line, Tumor; Disease Progression; Down-Regulation; Gene Expression Regulation, Neoplastic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Stomach Neoplasms; Tumor Microenvironment

Chronic infection with Helicobacter pylori increases risk of gastric diseases including gastric cancer. Despite development of a robust immune response, H. pylori persists in the gastric niche. Progression of gastric inflammation to serious disease outcomes is associated with infection with H. pylori strains which encode the cag Type IV Secretion System (cag T4SS). The cag T4SS is responsible for translocating the oncogenic protein CagA into host cells and inducing pro-inflammatory and carcinogenic signaling cascades. Our previous work demonstrated that nutrient iron modulates the activity of the T4SS and biogenesis of T4SS pili. In response to H. pylori infection, the host produces a variety of antimicrobial molecules, including the iron-binding glycoprotein, lactoferrin. Our work shows that apo-lactoferrin exerts antimicrobial activity against H. pylori under iron-limited conditions, while holo-lactoferrin enhances bacterial growth. Culturing H. pylori in the presence of holo-lactoferrin prior to co-culture with gastric epithelial cells, results in repression of the cag T4SS activity. Concomitantly, a decrease in biogenesis of cag T4SS pili at the host-pathogen interface was observed under these culture conditions by high-resolution electron microscopy analyses. Taken together, these results indicate that acquisition of alternate sources of nutrient iron plays a role in regulating the pro-inflammatory activity of a bacterial secretion system and present novel therapeutic targets for the treatment of H. pylori-related disease.

Topics: Animals; Disease Models, Animal; Epithelial Cells; Gastric Mucosa; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Immunity, Innate; Interleukin-8; Iron; Lactoferrin; Protein Isoforms; Type IV Secretion Systems

Nodular gastropathy (NG) is an inflammatory condition of the gastric mucosa characterized by the endoscopic detection of multiple millimeter protrusions. A strong association between NG and Helicobacter pylori and a possible role of NG as a risk factor for undifferentiated gastric cancer have been described. The aim of this study was to characterize the pathogenic and inflammatory profile of patients with NG.. Adult patients referred for upper gastrointestinal endoscopy were prospectively enrolled in this study. H. pylori infection status was determined by rapid urease test. Biopsies were stained with hematoxylin-eosin. Sydney and OLGA scores were used to assess gastritis characteristics and gastric cancer risk. PCR analysis was performed to determine bacterial load and virulence factors CagA (and its EPIYA motifs) and VacA alleles. Finally, gastric mucosa cytokine gene expression (IL-8, IL-1β, and TNF-α) was determined by real-time RT-PCR.. Forty-eight patients, mean age of 36 years, were recruited. All NG patients were infected by H. pylori. OLGA score was similar in both groups (NG patients and non-NG patients). NG patients had higher bacterial load in the gastric corpus (p = 0.01) and significantly less pro-inflammatory cytokine levels than non-NG infected patients (p = 0.01).. In our study, NG is not associated with preneoplastic lesions. An increase in bacterial load without a concomitant increase in mucosal inflammatory cytokine responses in H. pylori-infected subjects with NG may represent a general dampening of immune responses or an additional mechanism of H. pylori active immune evasion.

Topics: Adult; Antigens, Bacterial; Bacterial Load; Bacterial Proteins; Case-Control Studies; Cytokines; Endoscopy, Digestive System; Endosonography; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Narrow Band Imaging; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha; Young Adult

Bacterial outer membrane vesicles (OMVs) play a vital role in the mechanism of host-pathogen communication, while emerging evidence suggests that OMVs regulate host immune responses through differentially packaged small noncoding RNAs (sncRNAs) to target host mRNA function. Therefore, we identified differentially packaged sncRNAs in Helicobacter pylori OMVs and showed transfer of OMV sncRNAs to human gastric adenocarcinoma cells in this study. Our data revealed that sncRNAs (sR-2509025 and sR-989262) were enriched in OMVs, and reduced lipopolysaccharide or OMV-induced interleukin 8 (IL-8) secretion by cultured AGS cells. Collectively, these findings are consistent with the hypothesis that sncRNAs in H. pylori OMVs play a novel role in the mechanism of host-pathogen interaction, whereby H. pylori evades the host immune response.

Topics: Adenocarcinoma; Bacterial Outer Membrane Proteins; Cell Line; Cell Line, Tumor; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Immune Evasion; Interleukin-8; Protein Transport; RNA, Small Untranslated; Stomach Neoplasms

The increasing clinical significance of Helicobacter pylori (H. pylori) in human stomach cancer has led to global efforts to eradicate this pathogen. Recent studies have confirmed the importance of some cytokines such as Interleukin-18 (IL-18), Interleukin-8 (IL-8), Interleukin-17A (IL-17A) and Interleukin-22 (IL-22) in the pathogenesis of the so-called bacterium. This study was designed to compare the effects of Type 1T helper (Th1), Type 2T helper (Th2) cells, Regulatory T cells (Treg) and T helper 17 (Th17) modulatory effects on the efficacy of designed H. pylori vaccine by incorporating some molecular adjuvants in Treg competent and Treg suppressed groups. A bicistronic vector was used for simultaneous expression of codon-optimized Outer inflammatory protein a (OipA) gene and modified mice IL-18, IL-17A, IL-22 and Foxp3 (forkhead box P3) cytokines from four cassettes. Immunization of mice groups was performed using produced plasmids intradermally. Specific IgG1 and IgG2 and IgA antibody titers produced in mice were confirmed by enzyme-linked immunosorbent assay (ELISA) in sera and intestine obtained four weeks after the last immunization. After being stimulated with a mixture of both anti-CD28 mAb and H. pylori lysate, frequencies of single Interferon-Gamma (IFN-γ), single IL-17 and dual IFN-γ/IL-17-secreting T-cells were documented using dual-color FluoroSpot. The kinetics of Th1, Th2 and Th17 in the immunized animals was determined by relative quantification of IL-17A, IL-22, IFN-γ, IL-8, IL-2 and IL-4 specific mRNAs. Four weeks after bacterial challenge, quantitative colony count in the isolated and homogenized stomachs was utilized to assess the level of protective immunity among all groups. The results of immunologic assays showed that the highest cell-mediated immunity cytokines were produced in IL-17 receiving group in which the Treg responses were suppressed previously by the administration of the Foxp3 as an immunogen. In addition, potent clearance of Helicobacter pylori infection was seen in this group as well.

Topics: Adjuvants, Immunologic; Animals; Helicobacter Infections; Helicobacter pylori; Hepatocyte Nuclear Factor 3-gamma; Immunoglobulin G; Interferon-gamma; Interleukin-17; Interleukin-18; Interleukin-2; Interleukin-22; Interleukin-4; Interleukin-8; Interleukins; Mice; Recombinant Proteins; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells; Vaccines

Helicobacter pylori induces acute gastritis that can progress to serious diseases such as gastric cancer. H. pylori interacts with host cells within the gastric mucosa, resulting in activation of multiple innate immune signalling pathways, leading to pro-inflammatory cytokines production and immune cells recruitment. Various studies have shown that there are ethnic- and population-related differences in the expression of these cytokines. Although the H. pylori infection is a major public health problem in Morocco, to our knowledge, no study has been carried out in gastric cytokine expression from H. pylori-infected Moroccan patients. Thus we aimed to (i) determine the IL-1β, IL-8 and IL-17A gene expression in gastric biopsies from Moroccan patients infected with H. pylori, and (ii) to determine the cytokine signature of each pathological stages associated with this infection.. 71 patients with epigastralgic pain were included in this study. The H. pylori detection on gastric biopsies was performed by histopathological and PCR analysis. The IL-1β, IL-8 and IL-17A mRNA expression in the antrun and fundus biopsies was performed by RT-qPCR.. The histopathological and PCR analyses revealed that 87.32% of the patients were infected with H. pylori. IL-1β mRNA expression was significantly lower in the antral mucosa of H. pylori-infected patients (p = 0.0038) than in the uninfected while there was no significant difference in the expression of IL-8 and IL-17A mRNA. The expression of the three cytokines was higher in the fundic mucosa of H. pylori-infected patients than in the uninfected patients, but only IL-8 and IL-17A expression reached statistical significance (p = 0.042 and p = 0.0179 respectively). Furthermore, the multivariate predictive analysis highlighted a cytokine signature that may predict metaplasia during the infection progression that involves a specific down-regulation of IL17A and an up-regulation of IL1β in antral and fundic metaplasia respectively.

Topics: Adult; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Morocco; Signal Transduction; Stomach Neoplasms

Virulence factors of Helicobacter pylori (H. pylori) are diverse, so various biological responses happen in a host infected with H. pylori. The aim of this study is to conduct the metabolomics-based evaluation on H. pylori infection. AGS human gastric carcinoma cells were infected with H. pylori strain 26695, and then the altered metabolite pathways in the infected AGS cells were analyzed by metabolomics. Metabolites related to the glutathione (GSH) cycle were downregulated by H. pylori infection. Next, we evaluated the effects of H. pylori on the GSH-related pathway in AGS cells infected with H. pylori isolated from patients with atrophic gastritis (AG), duodenal ulcer (DU) and gastric cancer (GC). We found that the declined degree of GSH levels and oxidative stress were greater in AGS cells infected with GC strains than DU and AG-derived strains. There were no significant differences in almost mRNA expressions of GSH-related factors among different clinical strains, but the protein expression of glutathione synthetase was lower in AGS cells infected with GC-derived strains than DU and AG-derived strains. Our data demonstrates that GC-derived H. pylori-induced oxidative stress in a host is stronger and GC-derived strains may have suppressive influences on the host's GSH-related defense systems.

Topics: Cell Line, Tumor; Down-Regulation; Epithelial Cells; Glutathione; Glutathione Disulfide; Glutathione Synthase; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Metabolic Networks and Pathways; RNA, Messenger; Stomach; Virulence Factors

Aim To investigate the effects of interleukin-8 (IL-8) -251 T/A and +781 C/T polymorphism on the risk of Helicobacter pyloriinfection gastritis in children, and the IL-8 level of children with or without gastritis H. pylori infection according to polymorphism. Methods This prospective, case control clinical study included 64 children 2-18 years old. A disease group (32 gastritis patients with H. pylori-infection) was compared with a control group (32 gastritis patients without H. pylori infection). Demographic characteristics of patients were taken by a questionnaire; gastritis was confirmed by gastroscopy, H. pylori infection was confirmed with rapid urease test. Serum IL-8 level was measured by ELISA, and IL-8 -251 T/A and +781 C/T polymorphisms were analysed by RT-PCR. Demographic characteristics, IL-8 level, polymorphism of patients, and IL-8 level according to polymorphisms were compared between the groups. Results Children with tobacco exposure were associated with an increased risk of H. pylori-infection gastritis by 3.4-fold. There was a higher IL-8 level in the disease group compared to the control group. The disease group with IL-8 -251 AT polymorphism had a higher risk compared to TT polymorphism by 8.7-fold, and with IL-8 +781 CT polymorphism had a higher risk compared to CC polymorphism by 10.7-fold. Children in the disease group with IL-8 -251 AT and TT, and +781 CT and CC polymorphisms produced a higher IL-8 level than the control group in respective polymorphisms. Conclusion Children with H. pylori-infection gastritis have higher IL-8 production. There was an increased risk of developing H. pylori-infection in heterozygous -251 AT and +781 CT.

Topics: Adolescent; Case-Control Studies; Child; Child, Preschool; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Prospective Studies

Outer inflammatory protein (OipA) is an important virulence factor of Helicobacter pylori (H. pylori), but the correlation between oipA copy number and its virulence remains unknown. The study was designed to investigate whether the duplicate oipA gene loci showed more virulent than one oipA gene in vitro. H. pylori strain CCS9803 (China Chongqing Strain 9803) that carries duplicate oipA loci was used to construct one or two oipA knockout mutant strain, which was further verified by qPCR and western blot. Gastric epithelial cells AGS and GES-1 were infected with wild-type (WT) or oipA mutants for 6 or 24 h. The expression levels of IL-8, bacterial adhesion, cell apoptosis and cell cycle were performed to analyze the function of oipA. The WT and oipA mutant strains induce significantly higher mRNA and protein levels of IL-8 than the uninfected group (P < 0.05), but only oipA2 mutants induced significantly decreased expression levels than the WT-infected group (P < 0.05). Adherence to gastric cells was significantly decreased by inactivated two oipA loci (P < 0.05). The WT strain caused a significant rising proportion of early apoptosis cell, which had dropped after duplicate oipA genes were both knockout (P < 0.05). WT and oipA1 mutants failed to affect cell cycle; however, the oipA2 mutants increased M phase and reduced S phase when compared to the uninfected group. In conclusion, our study demonstrated that oipA impacts IL-8 expression, adherence, cell apoptosis and cell cycle of gastric cells independent of its gene copy number.

Topics: Apoptosis; Bacterial Adhesion; Bacterial Outer Membrane Proteins; Cell Cycle; Cells, Cultured; DNA Copy Number Variations; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Virulence; Virulence Factors

Topics: Carcinoma, Signet Ring Cell; Cell Line; Cell Transformation, Neoplastic; DNA, Bacterial; Exome Sequencing; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Kenya; Middle Aged; Phylogeny; Stomach Neoplasms; Virulence

Helicobacter pylori infection can cause a wide range of digestive diseases. Gene hp0788 encodes an outer membrane protein HofF, which can reduce the bacterial adherence to the GES-1 cells and affect pathogenesis of H. pylori. In this study, the role of hp0788 in H. pylori infection was further analyzed. RNA-seq data showed that two genes (hp0523 and hp0539), located on the cagPAI, were down-regulated in Δ0788 mutant. The changes were confirmed through qRT-PCR, and the expression of these two genes will be almost recovered to the normal level in complemented strain. These two genes, hp0523 and hp0539, are known to be necessary for integrated T4SS, which related to CagA translocation and IL-8 induction. In H. pylori infected assay, lower amount of phosphorylated CagA and lower induction of IL-8 were both detected in GES-1 cells infected by Δ0788 mutant, compared with the wild type strain. Meanwhile, these reductions almost recovered to the wild-type level in complemented strain. These results reveal that there is a correlation between hp0788 disruption and CagA/IL-8 decline. Deletion of CagA-encoding gene (hp0547) in Δ0788 mutant was further constructed. The double deleted mutant shows lower IL-8-inducing capability than Δ0788 mutant, indicated the correlation between deficiency of CagA and reduced IL-8 production. These results together imply that disruption of hp0788 might affect the efficiency of T4SS and CagA injection, then weaken the induction of IL-8 in infected GES-1 cells.

Topics: Antigens, Bacterial; Bacterial Adhesion; Bacterial Proteins; Cell Line, Tumor; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Stomach Diseases

CXC Chemokine Ligand 8 (CXCL8) plays an important role in gastric inflammation and in the progression of gastric cancer induced by Helicobacter pylori (H. pylori) infection. The association of CXCL8, CXC Chemokine Receptor 1 (CXCR1), and CXC Chemokine Receptor 2 (CXCR2) polymorphisms with H. pylori infection and gastric cancer progression needs to be investigated in a population within an enigma area consisting of multiple ethnicities, such as Thailand. To analyze the relative risk of H. pylori infection and gastric cancer among Thai gastroduodenal patients, gene polymorphisms in CXCL8 (promoter region -251) and in CXCR1 and CXCR2 (receptors for CXCL8) were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific-PCR (AS-PCR). We also determined the presence of cytotoxin-associated gene A (cagA) in Thai patients with H. pylori infection. Correlation between the CXCL8 (-251) polymorphism and CXCL8 gene expression was evaluated by quantitative reverse transcriptase-PCR (qRT-PCR). We found a significant association between the T/A and A/A genotypes of CXCL8 (-251) with H. pylori infection. However, no significant correlation was found between the CXCR1 (+2607) and CXCR2 (+1208) gene polymorphisms with H. pylori infection among Thai gastroduodenal subjects. Within the H. pylori-infected group of Thai gastroduodenal patients, no significant differences in cagA were observed. In addition, the A/A genotype of CXCL8 (-251) significantly correlated with the risk of gastric cancer and correlated with higher CXCL8 gene expression levels in Thai gastroduodenal patients. These results suggest that CXCL8 (-251) polymorphisms are associated with H. pylori infection, an increased risk of stronger inflammatory responses, and gastric cancer in Thai gastroduodenal patients.

Topics: Alleles; Disease Susceptibility; Female; Gastritis; Gene Frequency; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Polymorphism, Genetic; Population Surveillance; Stomach Neoplasms; Thailand

Inflammation induced by Helicobacter pylori infection related to gastric carcinogenesis. In this study, we have investigated the anti-inflammatory effect and its mechanism of kaempferol in the inflammatory response caused by H. pylori infection in vitro. We found that kaempferol reduced the expression of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-8) and production of IL-8 in AGS cells. In addition, kaempferol suppressed translocation of cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) of H. pylori to AGS cells. It was due to decreased transcription of type IV secretion system (T4SS) components involved in CagA injection and secretion system subunit protein A (SecA) of type V secretion system (T5SS) involved in VacA secretion by kaempferol. In conclusion, kaempferol shows the anti-inflammatory effect by suppressing the translocation of CagA and VacA proteins and leading to the down-regulation of pro-inflammatory cytokines. Abbreviations: CagA: cytotoxin-associated gene A; VacA: vacuolating cytotoxin A; T4SS: type IV secretion systems; SecA: secretion system subunit protein A; T5SS: type V secretion system.

Topics: Anti-Inflammatory Agents; Antigens, Bacterial; Bacterial Proteins; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Kaempferols; Protein Transport; Transforming Growth Factor alpha

Gastric acid secretion is compromised in chronic Helicobacter pylori (H. pylori) infection allowing overgrowth of non-H. pylori gastric bacteria (NHGB) in the stomach.. NHGB were isolated from gastric mucosa in selective media and further characterized with biochemical methods and 16S rRNA gene sequencing. Human gastric tissues were studied with indirect immunofluorescence with antibodies against H. pylori and Neisseria subflava (N. subflava). Gastric epithelial cell lines were cocultured with bacteria or incubated with lipopolysaccharides isolated from NHGB, and interleukin-8 released in the media was measured by enzyme-linked immunosorbent assay. Expression of Toll-like receptor (TLR)2, TLR4, it's coreceptor myeloid differentiation factor 2 (MD2), and CD14 in gastric cells was investigated by immunofluorescence microscopy and reverse transcriptase-polymerase chain reaction.. Haemophilus species, Neisseria species, Fusobacterium species, and Veillonella species were predominant Gram-negative bacteria coinfected with H. pylori. Lipopolysaccharides from N. subflava potently stimulated interleukin-8 secretion in MKN45 cells which was cancelled by preincubation with polymyxin B. TLR2, TLR4, CD14, and myeloid differentiation factor 2 were expressed in MKN45 cells, though their levels of expression were low. N. subflava adhered to MKN45 cells in vitro and colocalized with H. pylori in the human gastric mucosa.. Our data suggest that N. subflava colonized in the gastric mucosa contribute to gastric inflammation during chronic H. pylori gastritis.. NHGB may perpetuate gastric inflammation and accelerate neoplastic progression in the hypochlorhydric stomach.

Topics: Cell Line; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lipopolysaccharide Receptors; Lipopolysaccharides; Neisseria; Toll-Like Receptor 2; Toll-Like Receptor 4

Lack of a model that mirrors Helicobacter pylori-induced gastric mucosal inflammation has hampered investigation of early host-bacterial interactions. We used an ex vivo model of human stomach, gastric epithelial organoid monolayers (gastroid monolayers) to investigate interactions of H pylori infection and the apical junctional complex and interleukin-8 (IL-8) expression.. Morphology of human antral mucosal gastroid monolayers was evaluated using histology, immunohistochemical (IHC) staining, and transmission electron microscopy (TEM). Functional and gross changes in the apical junctional complexes were assessed using transepithelial electrical resistance (TEER), cytotoxicity assays, and confocal laser scanning microscopy. IL-8 expression was evaluated by real-time quantitative PCR and ELISA.. When evaluated by IHC and TEM, the morphology of gastroid monolayers closely resembled in vivo human stomach. Following inoculation of H pylori, TEER transiently declined (up to 51%) in an H pylori density-dependent manner. TEER recovered by 48 hours post-infection and remained normal despite continued presence and replication of H pylori. Confocal scanning microscopy showed minimal disruption of zonula occludens-1 or E-cadherin structure. IL-8 production was unchanged by infection with either CagA-positive or CagA-negative H pylori and JNK and MEK inhibitors did not suppress IL-8 production, whereas p38 and IKK inhibitor significantly did.. Human gastroid monolayers provide a model for experimental H pylori infection more consistent with in vivo human infections than seen with typical gastric epithelial cell lines. This ex vivo system should lead to better understanding of H pylori host-pathogen interactions.

Topics: Cell Line, Tumor; Cells, Cultured; Epithelial Cells; Gastritis; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Inflammation; Interleukin-8; Mutation; Stomach; Tight Junctions

C1q, as a LAIR-1 ligand, maintains monocytes quiescence and possess immunosuppressive properties. To understand the roles and molecular mechanisms, C1q mediated inflammation cytokines and several pivotal proteins in THP-1 cells after H. pylori infection were detected. The results showed that the expression of IL-8, IL-10, LAIR-1, phosphorylated/total JNK, phosphorylated/total p38-MAPK, phosphorylated/total AKT and phosphorylated/total NF-κB were up-regulated significantly in THP-1 cells after H. pylori infection. There was significant upregulation in IL-10 concentration, phosphorylated/total p38-MAPK and phosphorylated/total AKT, and downregulation in phosphorylated/total JNK in non-H. pylori infected THP-1 cells pretreated with C1q. C1q was also able to increase IL-8 and IL-10 production, and reduce LAIR-1 and phosphorylated/total p38-MAPK expression in pretreatment-C1q THP-1 cells after H. pylori infection. These results together indicated that H. pylori might induce IL-8 and IL-10 production through JNK, p38-MAPK, PI3K/AKT and NF-κB signaling pathway. C1q manipulate LAIR-1 to regulation IL-8 and IL-10 secretion in THP-1 cells after H. pylori infection through the p38-MAPK signaling pathway. This information is helpful to further understand the role and mechanisms of C1q on inflammation cytokines secretion in monocytes after H. pylori infection.

Topics: Complement C1q; Cytokines; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-8; MAP Kinase Signaling System; Monocytes; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phosphatidylinositol 3-Kinases; Phosphorylation; Receptors, Immunologic; Signal Transduction; THP-1 Cells; Up-Regulation

Type IV secretion systems are multiprotein complexes that mediate the translocation of macromolecules across the bacterial cell envelope. In Helicobacter pylori a type IV secretion system encoded by the cag pathogenicity island encodes 27 proteins and most are essential for virulence. We here present the identification and characterization of inhibitors of Cagα, a hexameric ATPase and member of the family of VirB11-like proteins that is essential for translocation of the CagA cytotoxin into mammalian cells. We conducted fragment-based screening using a differential scanning fluorimetry assay and identified 16 molecules that stabilize the protein suggesting that they bind Cagα. Several molecules affect binding of ADP and four of them inhibit the ATPase activity. Analysis of enzyme kinetics suggests that their mode of action is non-competitive, suggesting that they do not bind to the active site. Cross-linking suggests that the active molecules change protein conformation and gel filtration and transmission electron microscopy show that molecule 1G2 dissociates the Cagα hexamer. Addition of the molecule 1G2 inhibits the induction of interleukin-8 production in gastric cancer cells after co-incubation with H. pylori suggesting that it inhibits Cagα in vivo. Our results reveal a novel mechanism for the inhibition of the ATPase activity of VirB11-like proteins.

Topics: Adenosine Triphosphatases; Bacterial Proteins; Cell Line, Tumor; Enzyme Inhibitors; Helicobacter Infections; Helicobacter pylori; High-Throughput Screening Assays; Humans; Interleukin-8; Protein Conformation; Protein Multimerization; Stomach Neoplasms; Type IV Secretion Systems; Virulence

Helicobacter pylori (HP) is a pathogenic bacterium associated with chronic gastritis, gastric ulcer and gastric cancer. In the present study, the primary carcinogenesis process of normal gastric epithelial cells (GES‑1) infected with HP was investigated. It was determined that infected gastric mucosal epithelial GES‑1 cells secreted increased interleukin‑8 (IL‑8) and IL‑23, and exhibited enhanced expression of inducible nitric oxide synthase and cyclooxygenase‑2, inducing inflammatory reactions and resulting in apoptosis. The bacterial infection significantly increased the expression of carcinogenesis‑associated genes, including p16, c‑Myc, p53 and p21, as well as the expression of cell surface signaling molecules cluster of differentiation 44 (CD44) and CD54 in GES‑1 cells or tissues of patients with gastritis and gastric cancer in vitro or in vivo. Simultaneously, the migration and invasion abilities of normal gastric epithelial GES‑1 cells were increased following HP infection. These observations demonstrated that the inflammatory response of HP infection could cause normal gastric epithelial cells to undergo significant cancerous reactions, indicating that HP is a risk factor for gastric cancer.

Topics: Cell Line; Cell Movement; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-23; Interleukin-8; Neoplasm Invasiveness; Nitric Oxide Synthase Type II; Stomach; Stomach Neoplasms

CagL is an essential pilus surface component of the virulence-associated type IV secretion system (T4SS) employed by Helicobacter pylori to translocate the oncogenic effector protein CagA into human gastric epithelial cells. CagL contains an RGD motif and integrin α

Topics: Antigens, Bacterial; Antigens, Neoplasm; Bacterial Proteins; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Integrins; Interleukin-8; Neoplasms; Oligopeptides; Protein Binding; Protein Transport; Tumor Cells, Cultured; Type IV Secretion Systems

Helicobacter pylori infection occurs predominantly in childhood. Host immune response gene polymorphism is reported to affect the susceptibility to H pylori infection and the outcome of H pylori-related gastric cancer. Not all H pylori-infected patients, however, exhibit iron deficiency (ID). The relationship between host genetic polymorphisms and ID mediated by H pylori infection is not well understood.. Subjects (n = 644) from the general population of age 10 to 18 years were divided into 2 groups based on serology testing for anti-H pylori IgG: seropositive study group; and seronegative control group. Five single nucleotide polymorphisms (SNPs) in IL1B (rs1143627 and rs16944), IL8 (rs4073), IL10 (rs1800896), and ABO (rs505922), were genotyped and the iron status of the 2 groups was compared.. The seroprevalence rate for H pylori was 10.7% in this study. Infected subjects were significantly older and had lower serum iron levels than uninfected subjects (P = 0.0195 and 0.0059, respectively). Multivariate analysis revealed a significantly higher frequency of the T allele of rs505922 (odds ratio [OR] = 6.128; P < 0.001) and lower frequency of the T allele of rs1143627 (OR = 0.846; P = 0.014) in seropositive subjects. Among 59 seropositive subjects, the T allele frequency of rs1143627 was significantly higher in those with ID (OR = 3.156; P = 0.043), compared with those without ID.. ABO (rs505922) and IL1B (rs1143627) may affect H pylori infection susceptibility, and IL1B (rs1143627) may also influence ID risk in infected children.

Topics: ABO Blood-Group System; Adolescent; Anemia, Iron-Deficiency; Case-Control Studies; Child; Female; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-1beta; Interleukin-8; Iron; Male; Polymorphism, Single Nucleotide; Seroepidemiologic Studies; Taiwan

Helicobacter pylori (H. pylori) is the major stomach carcinogen, but the molecular mechanism responsible for the pathogenesis of cancer development mediated by H. pylori infection is still unclear. Aquaporin 3 (AQP3), overexpressed in gastric carcinoma, has a crucial role in gastric carcinogenesis and progression. However, the triggers and precise regulations for AQP3 upregulation during gastric carcinogens also remain unknown. Here we report that H. pylori infection-mediated carcinogenesis may be mechanistically depended on the upregulation of AQP3 expression via reactive oxygen species (ROS) pathway activation in the stomach. The retrospective analyses of clinical samples from patients with gastric carcinoma and other different stages of gastric diseases indicated that AQP3 expression was positively associated with gastric mucosal disease progression and H. pylori infection status as well. Furthermore, H. pylori infection significantly upregulated AQP3 and HIF-1α expression and increased ROS amount in human gastric epithelial AGS and GES-1 cells. Blockage of ROS with inhibitors, NAC and DPI, markedly decreased the expression of AQP3 and HIF-1α in both AGS and GES-1 cells simultaneously. Furthermore, the increased AQP3 in cells was mechanistically due to the transcriptional regulation by HIF-1α. In addition, H. pylori infection exerted production of proinflammatory cytokines IL-6, IL-8, and TNF depending on AQP3 level. Importantly, these in vitro novel findings were further investigated in vivo in a mouse H. pylori infectious model. Our current studies identify the mechanistic link between H. pylori infection and AQP3 upregulation in the pathogenesis of gastric carcinoma, which involves the activation of the ROS-HIF-1α axis and the exacerbated ROS-HIF-1α-AQP3-ROS loop.

Topics: Aged; Animals; Aquaporin 3; Cell Line, Tumor; Cell Transformation, Neoplastic; Epithelial Cells; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Interleukin-6; Interleukin-8; Male; Mice; Mice, Inbred C57BL; Middle Aged; Reactive Oxygen Species; Stomach Neoplasms; Tumor Necrosis Factor-alpha

Local Treg responses are involved in Helicobacter pylori-related inflammation and clinical outcomes after infection, and H. pylori-derived HSP60 (HpHSP60) is an important virulence factor associated with gastric carcinogenesis. This study to investigate the role of HpHSP60 in immunosuppression, particularly with regard to whether it could induce the production of Treg cells. For this purpose, human peripheral blood mononuclear cells (PBMCs) were treated with or without HpHSP60 in the presence of an anti-CD3 mAb to determine the effect of HpHSP60 on cell proliferation. In this report, HpHSP60 decreased the expression of CDK4 to significantly arrest the proliferation of mitogen-stimulated T-cells, which correlated with the induction of Treg cells. Moreover, monocytic cells were essential for the induction of HpHSP60-induced Treg cells via the secretion of IL-10 and TGF-β after treatment with HpHSP60. Blockage of HpHSP60 with specific monoclonal antibodies significantly reduced the colonization of H. pylori and the expression of Treg cells in vivo. Overall, our results suggest that HpHSP60 could act on macrophages to trigger the expression of IL-10 and TGF-β, thereby leading to an increase in Treg cells and inhibition of T-cell proliferation.

Topics: Animals; CD3 Complex; Cell Death; Cell Proliferation; Chaperonin 60; Cyclin-Dependent Kinase 4; Cytokines; Female; Gastric Mucosa; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Immunosuppression Therapy; Inflammation; Interleukin-10; Interleukin-8; Leukocytes, Mononuclear; Mice, Inbred BALB C; Monocytes; T-Lymphocytes, Regulatory; THP-1 Cells; Transforming Growth Factor beta; Virulence Factors

there are many determinant factors that may play roles in pathophysiology of functional dyspepsia. One of them is psychological stress that can increase plasma cortisol levels, alter inflammation process and affect Helicobacter pylori activity. No study has been conducted to find out the dominant factor among them. This study aimed to find the dominant factor among plasma cortisol levels, IL-6 and IL-8 expressions and H.Pylori activity, as the determinant factors in the pathophysiology of functional dyspepsia.. a cross-sectional study was conducted in 80 patients with dyspepsia syndrome at M. Djamil General Hospital, Padang, West Sumatera, Indonesia. The patients were categorized into two groups, i.e. the stress and non-stress group, which were identified using DASS 42 questionairre criteria. The inflammatory expressions (IL-6 and IL-8 expressions) as well as H. pylori activity were determined using immunohistochemistry of gastric biopsy specimens; while plasma cortisol levels was measured from peripheral blood samples. Data were analyzed using binary multivariate logistic regression.. there were 80 patients with functional dyspepsia with mean age of 38.9 years old. The morning cortisol levels was found significantly higher in the stress group. Higher IL-6 and IL-8 expressions were found in patients of non-stress group compared to those in the other group (IL-6: 73.28 (SD 16.60) vs. 72.95 (SD 19.49; and IL-8: 18.45 (SD 17.32) vs. 14.80 (SD 12.71)); although stastically not significant. There was greater Helicobacter pylori activity in the group with psychological stress compared to those in the non-stress group since there was antigen-antibody reaction invading the submucosa. The dominant determinant factor was the afternoon plasma cortisol levels.. many factors can become the determinant factors for gastric mucosal damage; however, our study has demonstrated that the dominant factor is afternoon plasma cortisol levels.

Topics: Adult; Cross-Sectional Studies; Dyspepsia; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Hydrocortisone; Immunohistochemistry; Indonesia; Interleukin-6; Interleukin-8; Logistic Models; Male; Middle Aged; Multivariate Analysis; Severity of Illness Index; Stress, Psychological

Helicobacter pylori infection has been reported to be associated with extra-digestive disorders such as respiratory diseases; however, the impact of H. pylori on lung is incompletely understood. Inflammatory response is mediated by the release of cytokines, interferon, and enzymes such as metalloproteinases (MMPs). This may contribute to collagen accumulation during the early phase of infection. MMP expression is an important factor for the proliferation and infiltration of lung cells in the process of fibrosis formation. The aim of this work was to study the impact of the infection with H. pylori on lung using a mouse model. We looked for histological lesions of lung infected with the microorganism as well as the expression of inflammatory and of endothelial dysfunction markers. C57BL/6 wild type (WT) mice were infected by orotracheal instillation with 20 μl of 1 × 10

Topics: Animals; Biomarkers; Bronchoalveolar Lavage; Catalase; Cytokines; Disease Models, Animal; DNA, Bacterial; Endothelial Cells; Gene Expression; Helicobacter Infections; Helicobacter pylori; Interleukin-10; Interleukin-1beta; Interleukin-4; Interleukin-6; Interleukin-8; Lung; Lung Injury; Male; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Nitric Oxide Synthase Type II; RNA, Ribosomal, 16S; Tumor Necrosis Factor-alpha

Topics: Adenosine Triphosphate; Antioxidants; Cell Line, Tumor; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Membrane Potential, Mitochondrial; Mitochondria; NF-kappa B; Oxidative Stress; PPAR gamma; Reactive Oxygen Species; Signal Transduction; Xanthophylls

The association between gastroesophageal reflux disease (GERD) prevalence and its risk factors in an area with low Helicobacter pylori prevalence is important to clarify. We analyzed the prevalence of GERD and risk factors in an area of Indonesia with low prevalence of H. pylori infection. We recruited 104 dyspeptic patients who underwent endoscopy in Surabaya. Patients were diagnosed with GERD based on the Los Angeles classification. We evaluated gastric biopsy specimens and measured serum pepsinogen levels. Interleukin polymorphisms were evaluated by polymerase chain reaction-restriction fragment length polymorphism. Of 104 patients, 56 (53.8%) were endoscopically found to have GERD, with most categorized as grade A; 48 (46.2%) were classified as non-GERD. Higher economic status, smoking, and a history of proton-pump inhibitor use significantly increased the risk of GERD. GERD Questionnaire scores showed a positive correlation with GERD (P < 0.001). An association was found between antral atrophic gastritis and GERD (P = 0.030), and patients with GERD more frequently had severe antral atrophy than nonerosive reflux disease (P = 0.018). We found an association between pepsinogen I/II levels and GERD (P = 0.047), but with low accuracy. IL-1β -511 TT and CT were predominant among the IL-1β -511 genotypes, and IL-8-251 AT and TT were predominant among the IL-8-251 genotypes. In conclusion, we found a high prevalence of GERD in an area with low prevalence of H. pylori infection, which could be associated with acid reflux. Smoking, history of proton-pump inhibitor use, and higher economic group significantly increased the risk of GERD.

Topics: Adolescent; Adult; Aged; Biopsy; Endoscopy; Female; Gastritis; Gastroesophageal Reflux; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Pepsinogen A; Polymorphism, Single Nucleotide; Risk Factors; Smoking; Young Adult

Helicobacter pylori virulence is associated with different clinical outcomes. The existence of an intact dupA gene from tfs4b cluster has been suggested as a predictor for duodenal ulcer development. However, the role of tfs plasticity zone clusters in the development of ulcers remains unclear. We studied several H. pylori strains to characterize the gene arrangement of tfs3 and tfs4 clusters and their impact in the inflammatory response by infected gastric cells.. The genome of 14 H. pylori strains isolated from Western patients, pediatric (n=10) and adult (n=4), was fully sequenced using the Illumina platform MiSeq, in addition to eight pediatric strains previously sequenced. These strains were used to infect human gastric cells, and the secreted interleukin-8 (IL-8) was quantified by ELISA. The expression of virB2, dupA, virB8, virB10, and virB6 was assessed by quantitative PCR in adherent and nonadherent fractions of H. pylori during in vitro co-infection, at different pH values.. We have found that cagA-positive H. pylori strains harboring a complete tfs plasticity zone cluster significantly induce increased production of IL-8 from gastric cells. We have also found that the region spanning from virB2 to virB10 genes constitutes an operon, whose expression is increased in the adherent fraction of bacteria during infection, as well as in both adherent and nonadherent fractions at acidic conditions.. A complete tfs plasticity zone cluster is a virulence factor that may be important for the colonization of H. pylori and to the development of severe outcomes of the infection with cagA-positive strains.

Topics: Adolescent; Adult; Bacterial Adhesion; Cells, Cultured; Child; Epithelial Cells; Female; Gastric Mucosa; Gene Expression Profiling; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Interleukin-8; Male; Middle Aged; Whole Genome Sequencing; Young Adult

Topics: Animals; Apoptosis; Cell Line, Tumor; Cell Survival; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Male; Mice; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Plant Extracts; Polygonum; Proto-Oncogene Proteins c-bcl-2; Quercetin; Rats; Seeds

Mannose-binding lectin (MBL) acts in the innate immune response to Helicobacter pylori. Interleukin 8 (IL-8) is a potent cytokine produced by gastric epithelial cells in response to H. pylori. We aimed to investigate whether polymorphisms in MBL2 and IL-8 influence susceptibility to H. pylori infection, and the associations of these polymorphisms with the risk of gastroduodenal diseases in a Korean population.. We consecutively enrolled 176 H. pylori-negative control subjects, 221 subjects with H. pylori-positive non-atrophic gastritis, 52 mild atrophic gastritis (AG), 61 severe AG, 175 duodenal ulcer, and 283 gastric cancer (GC). Allele-specific PCR-RFLP was conducted for polymorphisms in MBL2 exon 1 (codon 52, 54, and 57) and IL-8 -251 T > A. IL-8 levels in gastric mucosal tissues and serum MBL levels were measured by enzyme-linked immunosorbent assay.. MBL2 exon 1 polymorphic variants were found only in codon 54, and the allele frequencies did not differ significantly between the control and disease groups. Although serum MBL levels in codon 54 A/A mutants were markedly low, it did not influence susceptibility to H. pylori infection or the risk of gastroduodenal diseases. IL-8 levels were significantly different between T/T wild type, T/A heterozygote, and A/A mutant genotypes. IL-8 -251 A allele carriers (A/A + T/A) showed increased IL-8 levels, and were significantly associated with the risk of severe AG and GC.. We suggest that a combination of H. pylori infection and the IL-8 -251 T > A polymorphism might increase the risk of severe AG and GC in a Korean population.

Topics: Adult; Aged; Female; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Mannose-Binding Lectin; Middle Aged; Polymorphism, Single Nucleotide; Republic of Korea; Risk Factors; Stomach Neoplasms

Gastric mRNA expression of markers for acid secretion and inflammation and presence of gastric ulceration was studied in naturally Helicobacter suis-infected and non-infected 2-3 months old, 6-8 months old and adult pigs. In H. suis-infected 2-3 months old pigs, IL-8 and IL-1β transcript levels were upregulated in the pyloric gland zone, indicating an innate immune response. A similar response was demonstrated in the fundic gland zone of adult pigs, potentially due to a shift of H. suis colonization from the pyloric to the fundic gland zone. A Treg response in combination with decreased expressions of IL-8, IL-17A and IFN-γ was indicated to be present in the H. suis-infected 6-8 months old pigs, which may have contributed to persistence of H. suis. In H. suis-infected adult pigs, a Treg response accompanied by a Th17 response was indicated, which may have played a role in the decreased number of H. suis bacteria in the stomach of this age group. The decreased G-cell mass and upregulated expression of somatostatin indicated decreased acid secretion in H. suis-infected 6-8 months old pigs. In H. suis-infected adult pigs, upregulation of most markers for gastric acid secretion and increased G-cell mass was detected. Presence of severe hyperkeratosis and erosions in the non-glandular part of the stomach were mainly seen in the H. suis-positive groups. These results show that H. suis infection affects the expression of markers for acid secretion and inflammation and indicate that these effects differ depending on the infection phase.

Topics: Age Factors; Animals; Female; Gastric Acid; Gastric Mucosa; Gastritis; Helicobacter heilmannii; Helicobacter Infections; Interferon-gamma; Interleukin-17; Interleukin-1beta; Interleukin-8; Stomach; Swine; Swine Diseases

Helicobacter pylori is a Gram-negative, microaerophilic bacteria usually found in the stomach, which may evade its host's immune system and present long-term symptoms in affected individuals. This study aimed to evaluate the functional role of leukocyte-associated immunoglobulin (Ig)-like receptor-1 (LAIR-1) in the strategies and underlying molecular mechanisms by which H. pylori escapes the host's immune responses.. LAIR-1 knockdown THP-1 cells were used to detect cell apoptosis, cell proliferation, interleukin-8 (IL-8), IL-10, and activation of intracellular signaling induced by H. pylori.. Cell apoptosis, cell proliferation, IL-8, and IL-10 were increased in THP-1 cells after 24 h of H. pylori infection. Functional analysis indicated LAIR-1 silencing obviously inhibited the phosphorylation of IκBα, eIF2α, JNK, and Smad2 in the THP-1 after H. pylori infection. In addition, there were no significant differences in proliferation rates between control siRNA group and LAIR-1 siRNA group regardless of whether THP-1 cells were infected by H. pylori.. These results together indicated that LAIR-1 modulated cell apoptosis and inflammatory cytokines secretion in THP-1 cells, which might help sustain inflammation and prevent removal of the bacteria by the immune responses.

Topics: Apoptosis; Cell Proliferation; Cytokines; Gene Expression; Gene Knockdown Techniques; Gene Silencing; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Proteins; Interleukin-10; Interleukin-8; Monocytes; Phosphorylation; Receptors, Immunologic; RNA, Small Interfering; Signal Transduction; Smad2 Protein; THP-1 Cells

Cytotoxin-associated gene A (CagA) is one of the most important virulence factors of Helicobacter pylori, and serves a role in H. pylori‑mediated tumorigenesis in gastric cancer. However, the underlying molecular mechanism remains to be elucidated. The present study aimed to investigate the effects of CagA on the proliferation and apoptosis of GES‑1 cells, and the underlying mechanism. A CagA eukaryotic expression plasmid was constructed and transfected into GES‑1 cells. The mRNA and protein levels of CagA, tumor necrosis factor receptor‑associated factor 1 (TRAF1) and tumor necrosis factor receptor superfamily member 9 (4‑1BB) were determined using the reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. Western blot and ELISA analysis was used to detect the release of interleukin (IL)‑8. An MTT assay and flow cytometric analysis was used to assess cell viability and apoptosis, respectively. Ectopic expression of CagA markedly increased TRAF1 and 4‑1BB mRNA and protein levels in GES‑1 cells. CagA increased the expression and release of IL‑8 in GES‑1 cells. The expression of CagA significantly promoted the proliferation (P<0.05) and inhibited the apoptosis (P<0.05) of GES‑1 cells. In conclusion, CagA upregulated TRAF1/4‑1BB, thereby promoting the proliferation and inhibiting the apoptosis of GES-1 cells.

Topics: Antigens, Bacterial; Apoptosis; Bacterial Proteins; Cell Line; Cell Proliferation; Ectopic Gene Expression; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; TNF Receptor-Associated Factor 1; Transfection; Tumor Necrosis Factor Receptor Superfamily, Member 9

The human pathogen

Topics: Animals; Antigens, Bacterial; Bacterial Adhesion; Bacterial Proteins; Cell Line; Cell Membrane; Cholesterol; Cytokines; Epithelial Cells; Eukaryotic Cells; Female; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Membrane Microdomains; Membrane Proteins; Mice; Mice, Inbred C57BL; Mutagenesis; Mutation; RAW 264.7 Cells; Recombinant Proteins; Type IV Secretion Systems; Virulence

Highly virulent Helicobacter pylori cause proinflammatory signaling inducing the transcriptional activation and secretion of cytokines such as IL-8 in epithelial cells. Responsible in part for this signaling is the cag pathogenicity island (cagPAI) that codetermines the risk for pathological sequelae of an H. pylori infection such as gastric cancer. The Cag type IV secretion system (CagT4SS), encoded on the cagPAI, can translocate various molecules into cells, the effector protein CagA, peptidoglycan metabolites and DNA. Although these transported molecules are known to contribute to cellular responses to some extent, a major part of the cagPAI-induced signaling leading to IL-8 secretion remains unexplained. We report here that biosynthesis of heptose-1,7-bisphosphate (HBP), an important intermediate metabolite of LPS inner heptose core, contributes in a major way to the H. pylori cagPAI-dependent induction of proinflammatory signaling and IL-8 secretion in human epithelial cells. Mutants defective in the genes required for synthesis of HBP exhibited a more than 95% reduction of IL-8 induction and impaired CagT4SS-dependent cellular signaling. The loss of HBP biosynthesis did not abolish the ability to translocate CagA. The human cellular adaptor TIFA, which was described before to mediate HBP-dependent activity in other Gram-negative bacteria, was crucial in the cagPAI- and HBP pathway-induced responses by H. pylori in different cell types. The active metabolite was present in H. pylori lysates but not enriched in bacterial supernatants. These novel results advance our mechanistic understanding of H. pylori cagPAI-dependent signaling mediated by intracellular pattern recognition receptors. They will also allow to better dissect immunomodulatory activities by H. pylori and to improve the possibilities of intervention in cagPAI- and inflammation-driven cancerogenesis.

Topics: Adaptor Proteins, Signal Transducing; Antigens, Bacterial; Bacterial Proteins; Epithelial Cells; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Heptoses; Humans; Interleukin-8; Lipopolysaccharides; Protein Transport; Type IV Secretion Systems

Epidemiologic studies have reported an inverse relationship between childhood Helicobacter pylori infection and development of allergic asthma. Because lung epithelium plays an important role in allergic asthma pathogenesis, we hypothesized that H. pylori may directly influence airway epithelial cell innate immune function, particularly in early childhood. To test our hypothesis, we established an in vitro H. pylori infection model using primary tracheobronchial epithelial cell cultures derived from infant, juvenile and adult rhesus monkeys. Airway epithelial cell cultures were infected with wild-type or cag pathogenicity island mutant H. pylori strains, followed by evaluation of IL-8 and IL-6 protein synthesis. We found that H. pylori primarily increased IL-8 synthesis in a MOI and age-dependent fashion, with a greater than 4-fold induction in infant versus adult cultures. H. pylori-induced IL-8 synthesis in infant and juvenile cultures was significantly reduced by cag pathogenicity island mutants, indicating a requirement for the type IV secretion system. Although peptidoglycan recognition of nucleotide binding oligomerization domain-containing protein 1 (NOD1) and NF-kappaB have been implicated as key cytokine signaling molecules for H. pylori infection in gastric epithelium, NOD1 (ML130) or NF-kappaB (JSH-23) inhibitors minimally affected IL-8 synthesis in airway epithelial cell cultures following H. pylori infection. In contrast, inhibition of the p38 MAP kinase pathway (SB203580) resulted in almost complete suppression of H. pylori-induced IL-8 synthesis. Collectively, these results indicate that H. pylori can preferentially elicit IL-8 synthesis in a model of pediatric airway epithelium using the type IV secretion system via p38 MAP kinase.

Topics: Animals; Bacterial Proteins; Cell Line; Helicobacter Infections; Helicobacter pylori; Humans; In Vitro Techniques; Interleukin-6; Interleukin-8; p38 Mitogen-Activated Protein Kinases; Primates; Respiratory Mucosa; Signal Transduction; Type IV Secretion Systems

OipA is a phase-variable virulence factor of Helicobacter pylori. Mutations in oipA to turn the gene phase on in a cag pathogenicity island (PAI)-negative strain of H. pylori (J68) or phase off in a cag PAI-positive strain (26695) demonstrated that phase on oipA alleles in both strains had both increased oipA mRNA and human gastric adenocarcinoma (AGS) cell adherence compared to isogenic oipA phase off mutants. An oipA phase off mutant of H. pylori 26695 demonstrated decreased IL-8 secretion by AGS cells and failure to translocate the cag PAI effector CagA. Increased attachment by OipA expressing cag PAI-negative H. pylori J68 failed to alter secreted IL-8 levels. Thus, OipA is necessary but not sufficient for the induction of IL-8; however, it is necessary for translocation of the oncoprotein CagA. Perhaps the nearly invariant phase on status of oipA alleles among cag PAI-positive H. pylori isolates relates to the role of this outer membrane protein in effective translocation of CagA. oipA mRNA comparisons between AGS cell-adherent and non-adherent H. pylori 26695 revealed significantly greater levels in the adherent cells. This may allow H. pylori to adapt to conditions of host cell contact by altering expression of this virulence factor.

Topics: Amplified Fragment Length Polymorphism Analysis; Antigens, Bacterial; Bacterial Adhesion; Bacterial Outer Membrane Proteins; Bacterial Proteins; Cell Line, Tumor; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Protein Transport; RNA, Messenger; Type IV Secretion Systems

Gastric cancer risk is varied among different regions of Thailand. We examined the characteristics of Helicobacter pylori infection in two regions of Thailand. The H. pylori status of 273 dyspeptic patients (136 from the South and 137 from the North; a low and high incidence of gastric cancer region, respectively) was evaluated, and virulence genotypes (cagA, vacA, hrgA and jhp0562-positive/β-(1,3)galT) were determined. The overall H. pylori infection rate was 34.1% (93/273). The prevalence was higher in the North than in the South (50.4% vs. 17.6%, P <0.001) and was significantly higher among individuals with the following characteristics: low income, birthplace in the Northeast or North regions, agricultural employment, or consumption of alcohol or unboiling water. Among these socio-demographic determinants, region was an independent risk factor for H. pylori infection (odds ratio = 6.37). Patients including both H. pylori infected and uninfected cases who lived in the North had significantly more severe histological scores than those in the South. In contrast, among H. pylori-positive cases, patients in the South had significantly more severe histological scores than those in the North. Of the 74 strains cultured, 56.8% carried Western-type cagA, with a higher proportion in the South than in the North (76.2% vs. 49.1%, P = 0.05). In disagreement with the current consensus, patients infected with the Western-type cagA strains had more severe inflammation scores in the antrum than those infected with the East Asian-type cagA strains (P = 0.027). Moreover, Western-type cagA strains induced more severe histological scores in patients from the South than those of either genotype from the North. Other virulence genes had no influence on histological scores. The incidence of gastric cancer in Thailand was different among regions and corresponded to differences in the prevalence of H. pylori infection. More careful follow-up for patients in the South will be required, even if they are infected with H. pylori carrying Western-type cagA.

Topics: Cross-Sectional Studies; Gastric Mucosa; Genes, Bacterial; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Prevalence; Risk Factors; Stomach Neoplasms; Thailand; Virulence

Helicobacter pylori-induced gastric mucosal inflammation is mediated by proinflammatory and anti-inflammatory cytokines. Polymorphisms in genes that code cytokines influence cytokine secretion levels and appear to contribute to the risk of gastric diseases. In this sense, we performed this study to identify the polymorphisms in the IL-6, IL-8, and IL-10 genes and their associations with H. pylori infection and gastric pathologies.. Gastric biopsy samples of 151 patients infected with H. pylori and 76 uninfected individuals were used. Helicobacter pylori infection was diagnosed by histological examination and the detection of the ureA and glmM genes. The polymorphisms in the IL-6 (at position -174), IL-8 (at position -251), and IL-10 (at position -819) were detected by polymerase chain reaction-restriction fragment length polymorphism.. Among the genetic polymorphisms studied, we observed that only the presence of the A allele at position -251 of the IL-8 gene was significantly associated with H. pylori infection. In addition, patient carriers of the A/A genotype at position -251 of the IL-8 gene and carriers of the T allele at position -819 of the IL-10 gene had an increased risk of peptic ulcer disease in the presence of H. pylori infection. We did not find a correlation between polymorphisms in the IL-6, IL-8, and IL-10 genes and a higher risk of gastric carcinoma.. We demonstrated that polymorphisms in the IL-8 gene was significantly associated with H. pylori infection. Furthermore, polymorphisms in the IL-8 and IL-10 genes were associated with an enhanced risk of peptic ulcer disease in H. pylori-positive patients.

Topics: Biopsy; Female; Gastric Mucosa; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Interleukins; Male; Middle Aged; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Risk Factors; Stomach Neoplasms

Inflammatory signaling pathways induced by Helicobacter pylori remain unclear, having been studied mostly on cell-line models derived from gastric adenocarcinoma with potentially altered signaling pathways and nonfunctional receptors. Here, H. pylori-induced signaling pathways were investigated in primary human gastric epithelial cells. Inflammatory response was analyzed on chemokine mRNA expression and production after infection of gastric epithelial cells by H. pylori strains, B128 and B128Δ cagM, a cag type IV secretion system defective strain. Signaling pathway involvement was investigated using inhibitors of epidermal growth factor receptor (EGFR), MAPK, JAK and blocking Abs against TLR2 and TLR4. Inhibitors of EGFR, MAPK and JAK significantly reduced the chemokine mRNA expression and production induced by both H. pylori strains at 3 h and 24 h post-infection. JNK inhibitor reduced chemokine production at 24 h post-infection. Blocking Abs against TLR2 but not TLR4 showed significant reduction of chemokine secretion. Using primary culture of human gastric epithelial cells, our data suggest that H. pylori can be recognized by TLR2, leading to chemokine induction, and that EGFR, MAPK and the JAK/STAT signaling pathways play a key role in the H. pylori-induced CXCL1, CXCL5 and CXCL8 response in a cag pathogenicity island-independent manner.

Topics: Antibodies, Blocking; Cells, Cultured; Chemokine CXCL1; Chemokine CXCL5; Epithelial Cells; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation Mediators; Interleukin-8; Janus Kinases; Primary Cell Culture; Signal Transduction; Stomach; Toll-Like Receptor 2; Toll-Like Receptor 4

Multiple Helicobacter pylori strains colonize and coexist in the stomach of one single patient, carrying heterogeneous distributions of cag genotypes. The oesophagus provides a niche for H. pylori colonization; however, little is known about its adaptive role.. Using PCR for cagA, cagE and virB11 genes from cag-pathogenicity island (PAI) and Etest for antimicrobial susceptibility test, we determined cag-PAI genotypes associated with H. pylori virulence, when positive cultures were matching in both the stomach and the oesophagus (96 isolates; 8 out of 80 dyspeptic patients).. The stomach showed complete cag-PAI islands in 77 % of the isolates, whereas the oesophagus showed complete cag-PAI islands only in 44 % of the isolates. Expression of CagA and interleukin 8 correlated with inflammatory processes and histopathological changes in the stomach, but not in the oesophagus. Different cag-PAI profiles were found in both mucosae of an individual host, and at least one oesophagus profile corresponded to one profile identified in stomach. The antibiotic resistance profiles showed variability in the colonization by single or mixed H. pylori isolates in the gastric and oesophageal mucosa both intra- and inter-individuals.. These results demonstrate colonization with multiple H. pylori isolates in the oesophageal mucosa, like those found in the stomach of individual hosts. H. pylori was characterized by a dominant partial island, low interleukin 8 induction with lower histopathological damage and lower antibiotic resistance, suggesting that the microenvironmental changes in individual hosts select less virulent isolates in the oesophagus than in the stomach. New approaches to ensure effective eradication therapy in multi-resistant H. pylori strains must be developed.

Topics: Adult; Aged; Antigens, Bacterial; Bacterial Proteins; DNA, Bacterial; Esophagus; Female; Genomic Islands; Genotype; Genotyping Techniques; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Male; Middle Aged; Venezuela

Loss of Fas-associated factor 1 (FAF1) may act as a pro-survival signal in diseased cells, but whether this is true in gastric carcinoma remains unclear. Here we report that FAF1 was expressed at low levels in gastric carcinoma tissues and cell lines, and its expression correlated with larger tumors, higher histology grade, higher TNM stage, tumor infiltration, and lymph node metastasis. Univariate analysis and survival curve analysis identified low FAF1 expression as a predictor of poor prognosis. FAF1 overexpression in HGC-27 gastric cancer cells induced cell apoptosis and inhibited cell proliferation and growth. It also reduced colony formation in vitro and tumor growth in mice. We found that Helicobacter pylori, a risk factor for gastric cancer, down-regulated FAF1 expression via NF-κB signaling. Knock-down of IKKβ or p65 expression in gastric cancer cells reversed H. pylori-induced down-regulation of FAF1 expression and partially blocked H. pylori-induced secretion of inflammatory cytokines TNF-α and IL-8. Our results suggest that loss of FAF1 contributes to human gastric carcinogenesis by allowing H. pylori to activate NF-κB signaling.

Topics: Adaptor Proteins, Signal Transducing; Animals; Apoptosis; Apoptosis Regulatory Proteins; Carcinoma; Cell Line, Tumor; Cell Proliferation; Female; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; I-kappa B Kinase; Interleukin-8; Male; Mice, Nude; Middle Aged; NF-kappa B; Protein Interaction Maps; RNA Interference; Signal Transduction; Stomach Neoplasms; Time Factors; Transcription Factor RelA; Transfection; Tumor Burden; Tumor Necrosis Factor-alpha

Helicobacter pylori have colonized the gastric mucosa of half of the population worldwide. This bacterium is classified as a definitive type I carcinogen by the World Health Organization and no effective vaccine has been found against it yet. Thus, a logical and rational vaccine design against H. pylori is necessary. Because of its tremendous complexity and elicited immune responses, the vaccine design should considered multiple antigens to enhance immune-protection, involved in the different stages of pathogenesis besides inducing a specific immune response by B- and T-cell multi-epitopes. In this study, emphasis was placed on the design of a new unique vaccine named CTB-multiHp. In silico techniques were used to design a chimeric construct consisting of cholera toxin B subunit fused to multi-epitope of urease B (residue 148-158, 188-198), cytotoxin-associated gene A (residue 584-602), neutrophil activating protein (residue 4-28), vacuolating cytotoxin gene A (residue 63-81), H. pylori adhesine A (residue77-99), heat shock protein A (residue 32-54) and gamma glutamyl transpeptidase (residue 271-293). The tertiary structure and features of the vaccine were analyzed. The chimeric protein was expressed in Escherichia coli BL21 and the serology analyses indicated that the CTB-multiHp protein produced exhibit immune-reactivity. The results showed that CTB-multiHp could be a good vaccine candidate against H. pylori. Ongoing studies will evaluate the effects of CTB-multiHp against H. pylori infection.

Topics: Adhesins, Bacterial; Amino Acid Sequence; Antigens, Bacterial; Bacterial Proteins; Bacterial Vaccines; Cholera Toxin; Cloning, Molecular; Drug Design; Epitopes; Escherichia coli; gamma-Glutamyltransferase; Gene Expression; Heat-Shock Proteins; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Models, Molecular; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Fusion Proteins; Recombinant Proteins; Sequence Alignment; Urease

Adherence to the gastric epithelium is one of the most important steps of Helicobacter pylori to remain and cause disease. The aim of this study was to analyze whether H. pylori isolates from patients with different gastroduodenal diseases present differences in the pattern of adherence to gastric epithelial cells (AGS), in the ability to induce IL-8, and in the presence of virulence genes.. We tested 75 H. pylori strains isolated from nonatrophic gastritis, gastric cancer, and duodenal ulcer patients. The adhesion pattern and IL-8 induction were determined in AGS cells, and invasion of AGS cells was studied using a gentamicin protection assay. The IL-8 levels induced were determined by ELISA.. Helicobacter pylori strains presented diffuse adherence (DA) and localized (LA) adherence patterns, similar to those described for enteropathogenic E. coli (EPEC), were observed in AGS cells. A DA pattern was observed in 57% and LA in 43% of the strains, and DA was more frequent in isolates from patients with gastric cancer (p = 0.044). Strains with a LA pattern induced higher levels of IL-8 (p = 0.042) in AGS cells.. The adherence pattern was not associated with neither invasiveness nor with the presence of virulence genes. Our study shows that H. pylori strains present adherence patterns to AGS cells resembling those observed in EPEC and that these patterns may be associated with disease and with activity on AGS cells.

Topics: Bacterial Adhesion; Bacterial Proteins; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Middle Aged; Virulence Factors

Heterogeneity at the Helicobacter pylori cagA gene promoter region has been linked to variation in CagA expression and gastric histopathology. Here, we characterized the cagA promoter and expression in 46 H. pylori strains from Portugal. Our results confirm the relationship between cagA promoter region variation and protein expression originally observed in strains from Colombia. We observed that individuals with intestinal metaplasia were all infected with H. pylori strains containing a specific cagA motif. Additionally, we provided novel functional evidence that strain-specific sequences in the cagA promoter region and CagA expression levels influence interleukin 8 secretion by the host gastric epithelial cells.

Topics: Adult; Aged; Antigens, Bacterial; Bacterial Proteins; Case-Control Studies; Epithelial Cells; Female; Gene Expression; Genetic Variation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Portugal; Promoter Regions, Genetic; Young Adult

The purposes of this study were to examine the characteristics of Helicobacter pylori and the effect of Rumex Aquaticus Herba extract on the expression of cytokines in H. pylori-infected gastric epithelial cells. Cultured human adenocarcinoma gastric cells (AGS) were infected by H. pylori in RPMI 1640 media. Cell growth was measured by trypan blue assay. Western blot analysis was performed to investigate effect of extract containing Quercetin-3-O-β-d-glucuronopyranoside (ECQ) on the expression of inflammatory factors and the inhibition on cell growth. Furthermore, we compared the inhibitory effects with various combinations of clarithromycin, amoxicillin, omeprazole, and ECQ. The urease test with Christensen's Urea Agar was performed to identify the urease activity of H. pylori and the effect ECQ has on urease activity. When the cells were exposed to H. pylori, the trypan blue assay revealed a decrease in the rate of cell growth. Western blot analysis showed that H. pylori-infected cells had increased levels of degraded IκB-α and inflammatory factors. Pretreatment with ECQ inhibited interleukin expression induced by H. pylori in a dose-dependent manner. A combination of ECQ and antibiotics inhibited cytokine expression more effectively than other treatments. H. pylori displayed significant urease activity. ECQ did not significantly inhibit urease activity. These data suggest that H. pylori infection has cytotoxic effects against AGS cells, and ECQ may inhibit the production of proinflammatory cytokines in H. pylori-infected AGS cells.

Topics: Cell Line; Cell Proliferation; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Plant Extracts; Rumex

The epithelial cytokine response, associated with reactive oxygen species (ROS), is important in Helicobacter pylori (H. pylori)-induced inflammation. H. pylori induces the production of ROS, which may be involved in the activation of mitogen-activated protein kinases (MAPK), janus kinase/signal transducers and activators of transcription (Jak/Stat), and oxidant-sensitive transcription factor, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and thus, expression of interleukin-8 (IL-8) in gastric epithelial cells. α-lipoic acid, a naturally occurring thiol compound, is a potential antioxidant. It shows beneficial effects in treatment of oxidant-associated diseases including diabetes. The present study is purposed to investigate whether α-lipoic acid inhibits expression of inflammatory cytokine IL-8 by suppressing activation of MAPK, Jak/Stat, and NF-κB in H. pylori-infected gastric epithelial cells. Gastric epithelial AGS cells were pretreated with or without α-lipoic acid for 2 h and infected with H. pylori in a Korean isolate (HP99) at a ratio of 300:1. IL-8 mRNA expression was analyzed by RT-PCR analysis. IL-8 levels in the medium were determined by enzyme-linked immunosorbent assay. NF-κB-DNA binding activity was determined by electrophoretic mobility shift assay. Phospho-specific and total forms of MAPK and Jak/Stat were assessed by Western blot analysis. ROS levels were determined using dichlorofluorescein fluorescence. As a result, H. pylori induced increases in ROS levels, mRNA, and protein levels of IL-8, as well as the activation of MAPK [extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun NH2-terminal kinase 1/2 (JNK1/2), p38], Jak/Stat (Jak1/2, Stat3), and NF-κB in AGS cells, which was inhibited by α-lipoic acid. In conclusion, α-lipoic acid may be beneficial for prevention and/or treatment of H. pylori infection-associated gastric inflammation.

Topics: Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Janus Kinase 1; JNK Mitogen-Activated Protein Kinases; Mitogen-Activated Protein Kinases; NF-kappa B; Reactive Oxygen Species; RNA, Messenger; STAT3 Transcription Factor; Stomach; Thioctic Acid

Withaferin A (WA), a withanolide purified from Withania somnifera, has been known to exert anti-inflammatory effects. The present study sought to determine the effects of WA on Helicobacter (H.) pylori-mediated inflammation in the AGS gastric epithelial cell line. Cellular production of interleukin (IL)-8 and vascular endothelial growth factor (VEGF) was measured by ELISA. Western blot analysis was performed to determine the activation of nuclear factor (NF)-κB and mitogen-activated protein kinases (MAPKs) as well as hypoxia-inducible factor 1α stabilization. Bacterial growth was also examined by measuring the optical density. Pre-treatment or co-treatment with WA efficiently reduced IL-8 production by AGS cells in response to H. pylori infection. H. pylori-induced activation of NF-κB, but not MAPKs, was also inhibited by pre-treatment of WA in the cells. However, WA did not affect VEGF production and HIF-1α stabilization induced by H. pylori in AGS cells. In addition, WA did not influence the growth of H. pylori, suggesting that the anti-inflammatory effect of WA was not due to any bactericidal effect. These findings indicate that WA is a potential preventive or therapeutic agent for H. pylori-mediated gastric inflammation.

Topics: Epithelial Cells; Gastric Mucosa; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Inflammation; Interleukin-8; Mitogen-Activated Protein Kinases; NF-kappa B; Vascular Endothelial Growth Factor A; Withanolides

The aim of this study was to determine the biological function of hpsh4590 in Helicobacter pylori. After Hpsh4590 was expressed using a prokaryotic expression system, the cytotoxic effects and IL-8 production of Hpsh4590 were analyzed by co-culturing with GES-1 cells. Meanwhile, the antibody of rHpsh4590, produced by immunizing rabbit, was used for localization and protein interaction studies. Hpsh4590 fusion protein was expressed successfully in Escherichia coli Rosetta (DE3), and the polyclonal antibody was produced at high titers. The MTT assay showed that the inhibition ratio of GES-1 cells cultured with 0.1 μg/mL rHpsh4590 (3.02% ± 0.02%) was significantly lower than that of 20 μg/mL rHpsh4590 (57.57% ± 0.03%, p < 0.01), while DAPI staining showed the cytotoxic effects of rHpsh4590 for GES-1 cells. The up-regulation of cleaved caspase-3 and cleaved PARP was observed after GES-1 cells co-cultured with rHpsh4590 by Western blot. Co-culturing of GES-1 cells with rHpsh0459 (20 μg/mL) led to significant production of IL-8 at 12 h(1097.74 ± 212.37 pg/mL) and 24 h (1379.55 ± 209.58 pg/mL) then at 6 h(134.68 ± 14.64 pg/mL, p < 0.01). These observations suggest that the cytotoxicity of Hpsh4590 occurred in a concentration dependent manner, which is related with IL-8 secretion from gastric mucosal epithelial cells. Hpsh4590 was found localized in the membrane and the periplasm of H. pylori, interacted with zinc finger protein and methionine ABC transporter ATP-binding protein, and potentially regulates DNA uptake or transfer.

Topics: Animals; Bacterial Proteins; Caspase 3; Cell Membrane; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Protein Transport

Helicobacter pylori colonization of the human stomach can lead to adverse clinical outcomes including gastritis, peptic ulcers, or gastric cancer. Current data suggest that in addition to bacterial virulence factors, the magnitude and types of immune responses influence the outcome of colonization. Specifically, CD4+ T cell responses impact the pathology elicited in response to H. pylori. Because gastritis is believed to be the initiating host response to more detrimental pathological outcomes, there has been a significant interest in pro-inflammatory T cell cytokines, including the cytokines produced by T helper 17 cells. Th17 cells produce IL-17A, IL-17F, IL-21 and IL-22. While these cytokines have been linked to inflammation, IL-17A and IL-22 are also associated with anti-microbial responses and control of bacterial colonization. The goal of this research was to determine the role of IL-22 in activation of antimicrobial responses in models of H. pylori infection using human gastric epithelial cell lines and the mouse model of H. pylori infection. Our data indicate that IL-17A and IL-22 work synergistically to induce antimicrobials and chemokines such as IL-8, components of calprotectin (CP), lipocalin (LCN) and some β-defensins in both human and primary mouse gastric epithelial cells (GEC) and gastroids. Moreover, IL-22 and IL-17A-activated GECs were capable of inhibiting growth of H. pylori in vitro. While antimicrobials were activated by IL-17A and IL-22 in vitro, using a mouse model of H. pylori infection, the data herein indicate that IL-22 deficiency alone does not render mice more susceptible to infection, change their antimicrobial gene transcription, or significantly change their inflammatory response.

Topics: Animals; Anti-Infective Agents; CD4-Positive T-Lymphocytes; Epithelial Cells; Epithelium; Gastritis; Gastrointestinal Tract; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-17; Interleukin-22; Interleukin-8; Interleukins; Leukocyte L1 Antigen Complex; Lipocalins; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Real-Time Polymerase Chain Reaction; Stomach

Enterohepatic Helicobacter species are associated with several digestive diseases. Helicobacter pullorum is an emerging human foodborne pathogen, and Helicobacter hepaticus is a mouse pathogen; both species are associated with intestinal and/or hepatic diseases. They possess virulence factors, such as cytolethal distending toxin (CDT). Data indicate that CDT may be involved in chronic inflammatory responses, via its active subunit, CdtB. The proinflammatory properties of the CdtB of H. pullorum and H. hepaticus were assessed on human intestinal and hepatic epithelial cells in vitro. Interleukin 8 expression was evaluated by using wild-type strains and their corresponding CdtB isogenic mutants and by delivering CdtB directly into the cells. Nuclear factor κB nuclear translocation and transcriptomic characteristics in response to CdtB were also evaluated. The CdtB of these Helicobacter species induced nuclear factor κB nuclear translocation and exhibited proinflammatory properties, mainly the expression of T-helper type 17-related genes and genes encoding antimicrobial products also involved in cancer. The Histidine residue in position 265 of the CdtB catalytic site appeared to play a role in the regulation of most of these genes. As for flagellin or lipopolysaccharides, CdtB also induced expression of inflammation-associated genes related to antimicrobial activity.

Topics: Anti-Infective Agents; Bacterial Toxins; Cell Line, Tumor; Epithelial Cells; Gene Expression Profiling; Gene Expression Regulation; Helicobacter; Helicobacter Infections; Hepatocytes; Humans; Interleukin-8; Intestines; Mutation; Oligonucleotide Array Sequence Analysis; Th17 Cells; Virulence Factors

Galectin 3 (Gal-3) is upregulated in gastric epithelial cells as a host response to Helicobacter pylori infection. However, the significance of Gal-3 expression in H. pylori-infected cells is not well established. We analyzed Gal-3 intracellular expression, localization, and its effects in H. pylori-infected gastric epithelial cells. The predominantly nuclear confined Gal-3 was shown to be upregulated and exported out to the cytoplasm in H. pylori-infected AGS cells. The nuclear export was channeled through CRM-1 (exportin-1) protein. Interestingly, knock down of Gal-3 expression led to reduced NF-κB promoter activity and interleukin-8 (IL-8) secretion, suggesting its pro-inflammatory roles. Furthermore, Gal-3 was found to be pro-proliferative and anti-apoptotic in nature, as its knock down caused a reduction in cell proliferation and an increase in apoptosis, respectively. Taken together, our data suggest the expression and upregulation of Gal-3 as a critical endogenous event in H. pylori infection that interferes with various intracellular events, causing prolonged cell survival, which is characteristic in carcinogenesis.

Topics: Apoptosis Regulatory Proteins; Cell Line, Tumor; Cell Proliferation; Epithelial Cells; Exportin 1 Protein; Galectin 3; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Karyopherins; NF-kappa B; Protein Transport; Receptors, Cytoplasmic and Nuclear; Signal Transduction; Stomach Neoplasms; Up-Regulation

Topics: HEK293 Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Toll-Like Receptor 10; Toll-Like Receptor 2

Peptic ulcer disease and gastric cancer are caused most often by Helicobacter pylori strains that harbor the cag pathogenicity island, which encodes a type IV secretion system (T4SS) that injects the CagA oncoprotein into host cells. cagY is an essential gene in the T4SS and has an unusual DNA repeat structure that predicts in-frame insertions and deletions. These cagY recombination events typically lead to a reduction in T4SS function in mouse and primate models. We examined the role of the immune response in cagY-dependent modulation of T4SS function.. H pylori T4SS function was assessed by measuring CagA translocation and the capacity to induce interleukin (IL)8 in gastric epithelial cells. cagY recombination was determined by changes in polymerase chain reaction restriction fragment-length polymorphisms. T4SS function and cagY in H pylori from C57BL/6 mice were compared with strains recovered from Rag1-/- mice, T- and B-cell-deficient mice, mice with deletion of the interferon gamma receptor (IFNGR) or IL10, and Rag1-/- mice that received adoptive transfer of control or Ifng-/- CD4+ T cells. To assess relevance to human beings, T4SS function and cagY recombination were assessed in strains obtained sequentially from a patient after 7.4 years of infection.. H pylori infection of T-cell-deficient and Ifngr1-/- mice, and transfer of CD4+ T cells to Rag1-/- mice, showed that cagY-mediated loss of T4SS function requires a T-helper 1-mediated immune response. Loss of T4SS function and cagY recombination were more pronounced in Il10-/- mice, and in control mice infected with H pylori that expressed a more inflammatory form of cagY. Complementation analysis of H pylori strains isolated from a patient over time showed changes in T4SS function that were dependent on recombination in cagY.. Analysis of H pylori strains from mice and from a chronically infected patient showed that CagY functions as an immune-sensitive regulator of T4SS function. We propose that this is a bacterial adaptation to maximize persistent infection and transmission to a new host under conditions of a robust inflammatory response.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; CD4-Positive T-Lymphocytes; Cell Line; Chronic Disease; Epithelial Cells; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Homeodomain Proteins; Humans; Interferon gamma Receptor; Interferon-gamma; Interleukin-10; Interleukin-8; Mice; Mice, Inbred C57BL; Mice, Knockout; Receptors, Interferon; Recombination, Genetic; Signal Transduction; T-Lymphocytes, Helper-Inducer; Time Factors; Translocation, Genetic; Type IV Secretion Systems

Interleukin (IL)-8 is the key agent for initiating an inflammatory response to infection with Helicobacter pylori. Some strains of Lactobacillus spp. are known to colonize the stomach and suppress inflammation caused by H. pylori. In this study, we characterized two gastric-derived lactobacilli, Lactobacillus salivarius (LS) strains B37 and B60, capable of inhibiting H. pylori-induced IL-8 production by gastric epithelial cells.. Conditioned media from LS-B37 and LS-B60 suppressed H. pylori-induced IL-8 production and mRNA expression from AGS cells without inhibiting H. pylori growth. These conditioned media suppressed the activation of NF-κB but did not suppress c-Jun activation. IL-8 inhibitory substances in conditioned media of LS-B37 and LS-B60 are heat-stable and larger than 100 kDa in size. The inhibitory activity of LS-B37 was abolished when the conditioned medium was treated with α-amylase but still remained when treated with either proteinase K, trypsin, lipase or lysozyme. The activity of LS-B60 was abolished when the conditioned medium was treated with either amylase or proteinase K but still remained when treated with lysozyme. Treatment with lipase and trypsin also significantly affected the inhibitory activity of LS-B60 although the conditioned medium retained IL-8 suppression statistically different from media control.. These results suggest that L. salivarius strains B37 and B60 produce different immunomodulatory factors capable of suppressing H. pylori-induced IL-8 production from gastric epithelial cells. Our results suggest that the large, heat-stable immunomodulatory substance(s) present in the LCM of LS-B37 is a polysaccharide, while the one(s) of LS-B60 is either complex consisting of components of polysaccharide, lipid and protein or includes multiple components such as glycoprotein and lipoprotein.

Topics: alpha-Amylases; Anti-Inflammatory Agents; Cell Line; Culture Media, Conditioned; Endopeptidase K; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Lactobacillus; Ligilactobacillus salivarius; Lipase; Muramidase; NF-kappa B; Probiotics; RNA, Messenger; Stomach; Trypsin

The cytosolic host protein nucleotide binding oligomerization domain 1 (Nod1) has emerged as a key pathogen recognition molecule for innate immune responses in epithelial cells. The purpose of the study was to elucidate the mechanism by which Helicobacter pylori infection leads to transepithelial neutrophil migration in a Nod1-mediated manner.. Human epithelial cell lines AGS and Caco-2 were grown and infected with H. pylori. Interleukin (IL)-8 mRNA expression and IL-8 secretion were assessed, and nuclear factor κB (NF-κB) activation was determined. Stable transfections of AGS and Caco-2 cells with dominant negative Nod1 were generated. Neutrophil migration across the monolayer was quantified.. Cytotoxin-associated gene pathogenicity island (cagPAI)(+) H. pylori infection upregulated IL-8 mRNA expression and IL-8 secretion in AGS and Caco-2 cells compared with controls. NF-κB activation, IL-8 mRNA expression and IL-8 secretion by cagPAI knockdown strains were reduced compared with those infected with the wild-type strain. NF-κB activation, IL-8 mRNA expression and IL-8 secretion in dominant-negative (DN)-Nod1 stably transfected cells were reduced compared with the controls. The transepithelial migration of neutrophils in DN-Nod1 stably transfected cells was reduced compared with that in controls.. Signaling through Nod1 plays an essential role in neutrophil migration induced by the upregulated NF-κB activation and IL-8 expression in H. pylori-infected human epithelial cells.

Topics: Adult Stem Cells; Caco-2 Cells; Cell Line; Epithelial Cells; Gene Expression; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Neutrophils; NF-kappa B; Nod1 Signaling Adaptor Protein; RNA, Messenger; Signal Transduction; Transendothelial and Transepithelial Migration; Up-Regulation

The outcomes of Helicobacter pylori infection vary geographically. H pylori strains, disease presentation, and environments differ markedly in Bhutan and Dominican Republic. The aims were to compare the strains, histology, and expression of interleukin (IL) 8 and IL-10 from gastric mucosa from the 2 countries. H pylori status was assessed by the combination of rapid urease test, culture, and histology. Histology was evaluated using the updated Sydney System, and cytokines in gastric biopsies were measured using real-time polymerase chain reaction (PCR). There were 138 subjects from Bhutan and 155 from Dominican Republic. The prevalence of H pylori infection was 65% and 59%, respectively. The genotype of cagA was predominantly East Asian type in Bhutan versus Western type in Dominican Republic. Gastritis severity was significantly higher in H pylori-infected subjects from Bhutan than those from Dominican Republic. IL-8 expression by H pylori infection was 5.5-fold increased in Bhutan versus 3-fold in Dominican Republic (P < .001); IL-10 expression was similar. IL-8 expression levels among H pylori-infected cases tended to be positively correlated with polymorphonuclear leucocyte and monocyte infiltration scores in both countries. IL-8 expression among those with grade 2 and 3 polymorphonuclear leucocyte and monocyte infiltration was significantly higher in Bhutan than in Dominican Republic. The difference in IL-8 expression in the 2 countries is reflected in the different disease pattern between them. Whether the dominant factor is differences in H pylori virulence, in host-H pylori-environmental interactions, genetic factors or all remains unclear. However, severity of inflammation appears to be a critical factor in disease pathogenesis. We compared IL-8 messenger RNA levels between the high gastric cancer risk country, Bhutan (mainly East Asian-type H pylori), and the lower gastric cancer risk country, Dominican Republic (mainly Western-type H pylori).

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Bhutan; Biopsy; Dominican Republic; Environment; Female; Gastric Mucosa; Gastritis; Genetic Markers; Genotype; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Prevalence; Real-Time Polymerase Chain Reaction; Risk Factors; RNA, Messenger; Severity of Illness Index; Young Adult

Helicobacter pylori strains that harbor the oncoprotein CagA increase gastric cancer risk, and this risk is augmented under iron-deficient conditions. We demonstrate here that iron depletion induces coccoid morphology in strains lacking cagA. To evaluate the stability of augmented H. pylori virulence phenotypes stimulated by low-iron conditions, H. pylori isolated from iron-depleted conditions in vivo were serially passaged in vitro. Long-term passage decreased the ability of hypervirulent strains to translocate CagA or induce interleukin 8, indicating that hypervirulent phenotypes stimulated by low-level iron conditions are reversible. Therefore, rectifying iron deficiency may attenuate disease among H. pylori-infected persons with no response to antibiotics.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Gene Knockout Techniques; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Iron Deficiencies; Stomach; Virulence

Multiple studies have established the importance of the tol-pal gene cluster in bacterial cell membrane integrity and outer membrane vesicle (OMV) formation in Escherichia coli. In contrast, the functions of Tol-Pal proteins in pathogenic organisms, including those of the Epsilonproteobacteria, remain poorly if at all defined. The aim of this study was to characterize the roles of two key components of the Tol-Pal system, TolB and Pal, in OMV formation in the pathogenic bacterium, Helicobacter pylori.. H. pylori ΔtolB, Δpal and ΔtolBpal mutants, as well as complemented strains, were generated and assessed for changes in morphology and OMV production by scanning electron microscopy and enzyme-linked immunoassay (ELISA), respectively. The protein content and pro-inflammatory properties of OMVs were determined by mass spectroscopy and interleukin-8 (IL-8) ELISA on culture supernatants from OMV-stimulated cells, respectively.. H. pylori ΔtolB and Δpal bacteria exhibited aberrant cell morphology and/or flagella biosynthesis. Importantly, the disruption of H. pylori tolB but not pal resulted in a significant increase in OMV production. The OMVs from H. pylori ΔtolB and Δpal bacteria harbored many of the major outer membrane and virulence proteins observed in wild-type (WT) OMVs. Interestingly, ΔtolB, Δpal and ΔtolBpal OMVs induced significantly higher levels of IL-8 production by host cells, compared with WT OMVs.. This work demonstrates that TolB and Pal are important for membrane integrity in H. pylori. Moreover, it shows how H. pylori tolB-pal genes may be manipulated to develop "hypervesiculating" strains for vaccine purposes.

Topics: Bacterial Outer Membrane Proteins; Cell Membrane; Enzyme-Linked Immunospot Assay; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Mass Spectrometry; Microscopy, Electron, Scanning; Periplasmic Proteins

Cinnamomum cassia is widely employed for gastrointestinal complaints such as dyspepsia, flatulence, diarrhea, and vomiting. Studies report cinnamaldehyde (CM) as a major active constituent of cinnamon. The aim of this study was to evaluate the anti-inflammatory mechanism of CM on Helicobacter (H.) pylori-infected gastric epithelial cells in order to validate cinnamon traditional use in gastrointestinal (GI)-related disorders. AGS/MKN-45 cells and H. pylori (193C) were employed for co-culture experiments. Anti-H. pylori cytotoxic and anti-adhesion activity of CM were determined. Enzyme linked immunosorbent assay, real time polymerase chain reaction analysis and immunoblotting were used to measure the effect on interleukin-8 (IL-8) secretion/expression. The effect on activation of nuclear factor kappa B (NF-κB) was determined by immunoblot analysis. The non-cytotoxic CM (≤125 µM) was also non-bactericidal at the given time, suggesting the effect in H. pylori/cell co-culture system was not due to alteration in H. pylori viability or the toxicity to the cells. Also, CM did not show any anti-adhesion effect against H. pylori/cell co-culture. However, pre-incubation of the cells with CM significantly inhibited the IL-8 secretion/expression from H. pylori-infected cells (p<0.01). In addition, CM suppressed H. pylori-induced NF-κB activation and prevented degradation of inhibitor (I)-κB This study provides evidence that the anti-inflammatory effect of C. cassia on H. pylori-infected gastric cells is due to blockage of the NF-κB pathway by cinnamaldehyde. This agent can be considered as a potential candidate for in vivo and clinical studies against various H. pylori related gastric pathogenic processes.

Topics: Acrolein; Anti-Inflammatory Agents; Cell Line; Cell Line, Tumor; Epithelial Cells; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B

Topics: Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male

Topics: Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male

Pathogenicity of the human pathogen Helicobacter pylori relies on its capacity to adapt to a hostile environment and to escape the host response. Although there have been great advances in our understanding of the bacterial cytoskeleton, major gaps remain in our knowledge of its contribution to virulence. In this study we have explored the influence of coiled coil rich proteins (Ccrp) cytoskeletal elements on pathogenicity factors of H. pylori. Deletion of any of the ccrp resulted in a strongly decreased activity of the main pathogenicity factor urease. We further investigated their role using in vitro co-culture experiments with the human gastric adenocarcinoma cell line AGS modeling H. pylori - host cell interactions. Intriguingly, host cell showed only a weak "scattering/hummingbird" phenotype, in which host cells are transformed from a uniform polygonal shape into a severely elongated state characterized by the formation of needle-like projections, after co-incubation with any ccrp deletion mutant. Furthermore, co-incubation with the ccrp59 mutant resulted in reduced type IV secretion system associated activities, e.g. IL-8 production and CagA translocation/phosphorylation. Thus, in addition to their role in maintaining the helical cell shape of H. pylori Ccrp proteins influence many cellular processes and are thereby crucial for the virulence of this human pathogen.

Topics: Antigens, Bacterial; Bacterial Proteins; Cell Line; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Mutation; Phenotype; Type IV Secretion Systems; Urease; Virulence; Virulence Factors

Helicobacter pylori (H. pylori) induces the activation of nuclear factor-kB (NF-κB) and cytokine expression in gastric epithelial cells. The Janus kinase/signal transducers and activators of transcription (Jak/Stat) cascade is the inflammatory signaling in various cells. The purpose of the present study is to determine whether H. pylori-induced activation of NF-κB and the expression of interleukin-8 (IL-8) are mediated by the activation of Jak1/Stat3 in gastric epithelial (AGS) cells. Thus, gastric epithelial AGS cells were infected with H. pylori in Korean isolates (HP99) at bacterium/cell ratio of 300:1, and the level of IL-8 in the medium was determined by enzyme-linked immonosorbent assay. Phospho-specific and total forms of Jak1/Stat3 and IκBα were assessed by Western blot analysis, and NF-κB activation was determined by electrophoretic mobility shift assay. The results showed that H. pylori induced the activation of Jak1/Stat3 and IL-8 production, which was inhibited by a Jak/Stat3 specific inhibitor AG490 in AGS cells in a dose-dependent manner. H. pylori-induced activation of NF-κB, determined by phosphorylation of IκBα and NF-κB-DNA binding activity, were inhibited by AG490. In conclusion, Jak1/Stat3 activation may mediate the activation of NF-κB and the expression of IL-8 in H. pylori-infected AGS cells. Inhibition of Jak1/Stat3 may be beneficial for the treatment of H. pylori-induced gastric inflammation, since the activation of NF-κB is inhibited and inflammatory cytokine expression is suppressed.

Topics: Blotting, Western; DNA, Bacterial; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Janus Kinase 1; NF-kappa B; Phosphorylation; RNA, Messenger; Signal Transduction; STAT3 Transcription Factor

Helicobacter pylori (H. pylori) infection plays an important role in the carcinogenesis and development of gastric cancer. Eradication of H. pylori can effectively reduce the risk of gastric cancer, but the underlying mechanisms are not fully understood. This study aimed to investigate the effect of eradication of H. pylori on the expression levels of FHIT, IL-8 and P73 in the gastric mucosa of first-degree relatives of gastric cancer patients.. One hundred and thirty-two patients with functional dyspepsia having first-degree relatives with gastric cancer were prospectively recruited in this study. Nine patients presented with H. pylori infection and family histories of gastric cancer, 61 with H. pylori infection and without family histories of gastric cancer, 6 without H. pylori infection and with family histories of gastric cancer, and 56 without H. pylori infection and family histories of gastric cancer. The protein and mRNA expression levels of FHIT, IL-8 and P73 in gastric mucosa of the subjects were detected by immunohistochemical staining and polymerase chain reaction, respectively.. Compared with the patients without H. pylori infection and family histories of gastric cancer, both the protein and mRNA levels of FIHT significantly decreased in patients with H. pylori infection and/or family histories of gastric cancer, and both the protein and mRNA levels of IL-8 significantly increased. After eradication of H. pylori, both the protein and mRNA levels of FHIT were significantly higher, and both the protein and mRNA levels of IL-8 were significantly lower. However, H. pylori infection and family histories of gastric cancer had no major effect on P73 expression.. Down-regulation of FHIT and up-regulation of IL-8 may be involved in the pathogenesis of H. pylori infection in the first-degree relatives of gastric cancer patients.

Topics: Acid Anhydride Hydrolases; Adult; Aged; Anti-Bacterial Agents; Demography; DNA-Binding Proteins; Down-Regulation; Dyspepsia; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Neoplasm Proteins; Nuclear Proteins; Proton Pump Inhibitors; Real-Time Polymerase Chain Reaction; RNA, Messenger; Stomach Neoplasms; Tumor Protein p73; Tumor Suppressor Proteins; Up-Regulation

Acute Helicobacter pylori infection of gastric epithelial cells and human gastric biopsies represses H,K-ATPase α subunit (HKα) gene expression and inhibits acid secretion, causing transient hypochlorhydria and supporting gastric H. pylori colonization. Infection by H. pylori strains deficient in the cag pathogenicity island (cag PAI) genes cagL, cagE, or cagM, which do not transfer CagA into host cells or induce interleukin-8 secretion, does not inhibit HKα expression, nor does a cagA-deficient strain that induces IL-8. To test the hypothesis that virulence factors other than those mediating CagA translocation or IL-8 induction participate in HKα repression by activating NF-κB, AGS cells transfected with HKα promoter-Luc reporter constructs containing an intact or mutated NF-κB binding site were infected with wild-type H. pylori strain 7.13, isogenic mutants lacking cag PAI genes responsible for CagA translocation and/or IL-8 induction (cagA, cagζ, cagε, cagZ, and cagβ), or deficient in genes encoding two peptidoglycan hydrolases (slt and cagγ). H. pylori-induced AGS cell HKα promoter activities, translocated CagA, and IL-8 secretion were measured by luminometry, immunoblotting, and ELISA, respectively. Human gastric biopsy acid secretion was measured by microphysiometry. Taken together, the data showed that HKα repression is independent of IL-8 expression, and that CagA translocation together with H. pylori transglycosylases encoded by slt and cagγ participate in NF-κB-dependent HKα repression and acid inhibition. The findings are significant because H. pylori factors other than CagA and IL-8 secretion are now implicated in transient hypochlorhydria which facilitates gastric colonization and potential triggering of epithelial progression to neoplasia.

Topics: Achlorhydria; Antigens, Bacterial; Bacterial Proteins; Cells, Cultured; Epithelial Cells; Gastric Acid; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Promoter Regions, Genetic; Proton Pumps; Signal Transduction; Sodium-Potassium-Exchanging ATPase; Virulence Factors

to investigate the serum levels of TNF-a, IL-8, VEGF in Helicobacter pylori infection, and their association with the degrees of gastritis histopathology.. a cross-sectional study was done on 80 consecutive gastritis patients admitted to endoscopy units at Adam Malik General Hospital and Permata Bunda Hospital, Medan, Indonesia from July-December 2014. The Rapid Urease test was used for the diagnosis of H. pylori infection. The severity of chronic inflammation, neutrophil infiltration, atrophy, and intestinal metaplasia were assessed. Serum samples were obtained to determine circulating TNF-a, IL-8, and VEGF. Univariate and bivariate analysis (chi square, fisher's exact, and mann-whitney test) were done using SPSS version-22.. there were 41.25% of 80 patients infected with Helicobacter pylori. Serum TNF-a and VEGF levels in the infected group were significantly higher compared to H. pylori negative, but there were no significant differences between serum levels of IL-8 in H. pylori positive and negative. There were significant associations between serum level of TNF-a and IL-8 with degree of chronic inflammation, and also between serum level of IL-8 and degree of neutrophil infiltration. There were significant associations between serum level of VEGF and degree of atrophy, and also between serum level of VEGF and degree of intestinal metaplasia.. High levels of TNF-a were associated with severe degree of chronic inflammation, high levels of IL-8 associated with severe degree of chronic inflammation and neutrophil infiltration, and high levels of VEGF associated with severe degree of premalignant gastric lesion.

Topics: Adult; Cross-Sectional Studies; Endoscopy, Gastrointestinal; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Indonesia; Inflammation; Interleukin-8; Male; Metaplasia; Middle Aged; Neutrophil Infiltration; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

The aim of this conventional case-control study was to investigate the prevalence and relationship between Helicobacter pylori infection in type 2 diabetes mellitus (DM) and diabetic nephropathy (DN).. A total of 241 type 2 DM patients and 69 non-diabetic subjects with dyspeptic symptoms were enrolled in the study. Gastroduodenal lesions were observed by gastrointestinal endoscopy and the presence of H. pylori was identified by rapid urease test and serum IgG antibodies to H. pylori. According to the urinary albumin excretion rate (UAE), patients were classified into diabetes mellitus group (DM group, with UAE <30 mg/24h); diabetic nephropathy group 1 (DN group 1, with UAE 30 mg/24 h to <300 mg/24 h); and diabetic nephropathy group 2 (DN group 2 ≥ 300 mg/24 h). The 69 non-diabetic subjects were used as control group. The serum levels of inflammatory factors such as tumor necrosis factor-α (TNF-α) and interleukin (IL)-8 were determined using ELISA.. The prevalence of H. pylori infection in DN group 1 and DN group 2 was 45/72 (62.5%) and 34/53 (64.15%), respectively, which was significantly higher than in control [28/65 (43.1%)] and DM groups [42.9% (27/63)]. No significant differences of H. pylori prevalence were detected between DN groups as well as DM and control groups. Interestingly, in both DN groups, higher levels of IL-8, TNF-α and urinary albumin excretion rate were found in H. pylori positive subjects.. Diabetic nephropathy patients are more susceptible to H. pylori infection. Our data support an association between H. pylori infection and diabetic nephropathy.

Topics: Adult; Albuminuria; Case-Control Studies; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Prevalence; Tumor Necrosis Factor-alpha

Helicobacter pylori (H. pylori)-induced oxidative stress has been shown to play a very important role in the inflammation of the gastric mucosa and increases the risk of developing gastric cancer. Resveratrol has many biological functions and activities, including antioxidant and anti-inflammatory effect. The purpose of this study was to probe whether resveratrol inhibits H. pylori-induced gastric inflammation and to elucidate the underlying mechanisms of any effect in mice. A mouse model of H. pylori infection was established via oral inoculation with H. pylori. After one week, mice were administered resveratrol (100 mg/kg body weight/day) orally for six weeks. The mRNA and protein levels of iNOS and IL-8 were assessed using RT-PCR, Western blot and ELISA. The expression levels of IκBα and phosphorylated IκBα (which embodies the level and activation of NF-κB), Heme Oxygenase-1 (HO-1; a potent antioxidant enzyme) and nuclear factor-erythroid 2 related factor 2 (Nrf2) were determined using Western blot, and lipid peroxide (LPO) level and myeloperoxidase (MPO) activity were examined using an MPO colorimetric activity assay, thiobarbituric acid reaction, and histological-grade using HE staining of the gastric mucosa. The results showed that resveratrol improved the histological infiltration score and decreased LPO level and MPO activity in the gastric mucosa. Resveratrol down-regulated the H. pylori-induced mRNA transcription and protein expression levels of IL-8 and iNOS, suppressed H. pylori-induced phosphorylation of IκBα, and increased the levels of HO-1 and Nrf2. In conclusion, resveratrol treatment exerted significant effects against oxidative stress and inflammation in H. pylori-infected mucosa through the suppression of IL-8, iNOS, and NF-κB, and moreover through the activation of the Nrf2/HO-1 pathway.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Disease Models, Animal; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Heme Oxygenase-1; Interleukin-8; Lipid Peroxides; Male; Mice; NF-kappa B; Nitric Oxide Synthase Type II; Oxidative Stress; Peroxidase; Phosphorylation; Resveratrol; Stilbenes

Helicobacter pylori cagPAI genes play an important role in pathogenesis, however little is known about their functions in isolates from Turkish patients. We aimed to evaluate the intactness and the effect of the cagPAI genes (cagT, cagM, cagE, cagA) and cagA EPIYA motifs on the AGS morphological changes and IL-8 induction. Of 53 patients 38 were found infected with H. pylori. PCR amplification of the cagPAI genes showed 42.1 % intact, 39.5 % partially deleted and 18.4 % with complete deletions. Isolates from gastritis, duodenal and gastric ulcer patients with intact and partially deleted cagPAI genes induced higher IL-8 secretion than those with complete deletions. Isolates from gastritis patients had higher deletion frequencies of the cagT and cagM genes than the other two genes. Infection of AGS cells with isolates that possess intact cagPAI and EPIYA-ABC resulted in the formation of the hummingbird phenotype. The cagA positive isolates induced higher IL-8 secretion than cagA negative isolates. Isolates from DU patients with more than one EPIYA-C motif induced higher concentrations of IL-8 than those with EPIYA-ABC. In conclusion, the intactness of the cagPAI in our isolates from different patients was not conserved. An intact cagPAI was found to play an important role in the pathogenesis of DU but not GU or gastritis. The cagA gene, but not other cagPAI genes, was associated with the induction of IL-8 and the morphological changes of the AGS cells. An increase in the number of EPIYA-C motifs had noticeable effect on the formation of the hummingbird phenotype.

Topics: Adult; Aged; Antigens, Bacterial; Bacterial Proteins; Epithelial Cells; Female; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Middle Aged; Virulence Factors; Young Adult

The envisaged roles and partly understood functional properties of Helicobacter pylori protein HP0986 are significant in the context of proinflammatory and or proapoptotic activities, the two important facilitators of pathogen survival and persistence. In addition, sequence analysis of this gene predicts a restriction endonuclease function which remained unknown thus far. To evaluate the role of HP0986 in gastric inflammation, we studied its expression profile using a large number of clinical isolates but a limited number of biopsies and patient sera. Also, we studied antigenic role of HP0986 in altering cytokine responses of human gastric epithelial (AGS) cells including its interaction with and localization within the AGS cells.. For in vitro expression study of HP0986, 110 H. pylori clinical isolates were cultured from patients with functional dyspepsia. For expression analysis by qRT PCR of HP0986, 10 gastric biopsy specimens were studied. HP0986 was also used to detect antibodies in patient sera. AGS cells were incubated with recombinant HP0986 to determine cytokine response and NF-κB activation. Transient transfection with HP0986 cloned in pEGFPN1 was used to study its subcellular localization or homing in AGS cells.. Out of 110 cultured H. pylori strains, 34 (31%) were positive for HP0986 and this observation was correlated with in vitro expression profiles. HP0986 mRNA was detected in 7 of the 10 biopsy specimens. Further, HP0986 induced IL-8 secretion in gastric epithelial cells in a dose and time-dependent manner via NF-κB pathway. Serum antibodies against HP0986 were positively associated with H. pylori positive patients. Transient transfection of AGS cells revealed both cytoplasmic and nuclear localization of HP0986.. HP0986 was moderately prevalent in clinical isolates and its expression profile in cultures and gastric biopsies points to its being naturally expressed. Collective observations including the induction of IL-8 via TNFR1 and NF-κB, subcellular localization, and seropositivity data point to a significant role of HP0986 in gastroduodenal inflammation. We propose to name the HP0986 gene/protein as 'TNFR1 interacting endonuclease A (TieA or tieA)'.

Topics: Antigens, Bacterial; Biopsy; Dyspepsia; Epithelial Cells; Female; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Male; Middle Aged; NF-kappa B; Receptors, Tumor Necrosis Factor, Type I; Virulence Factors

TLRs play an essential role in the initial handling of H. pylori and determine the clinical outcomes that range from simple asymptomatic gastritis to peptic ulcer disease and gastric cancer. Asp299Gly and Thr399Ile polymorphisms in TLR4 have been associated with a variety of inflammatory and infectious conditions including gastric cancer. The T-251A polymorphism in the promoter region of IL-8 gene has been found to be associated with changing the in vitro levels of IL-8 production. IL-8 exhibits angiogenic activity and is responsible for tumor-associated angiogenesis in several cancers.. 130 gastric cancer patients and 200 healthy controls were included in this study. DNA extraction was followed by PCR detection of H. pylori infection, PCR-RFLP for the TLR 4 polymorphism and PCR-CTPP for IL-8 gene polymorphism.. The adjusted OR for gastric cancer risk was 1.15 (95% CI, 0.8357-1.3463); 1.39 (0.6964-2.781) and 1.43 (0.954-2.1515) for Asp299Gly, Thr399Ile and IL-8 T_251A respectively. Odds Ratio analysis showed CT genotype and AT and AA genotypes as risk factors for the development of gastric cancer. We found the Asp299Gly polymorphism carrier to be significantly associated (p value 0.03)with the development of tumours in the distal part of the stomach and Thr399Ile polymorphism to be significantly associated(p value 0.008) with the development of well-differentiated gastric adenocarcinoma.The IL-8 T-251A polymorphism was not found to be associated with any of the clinicopathological characteristics.. No correlation was found between the appearance of disease and HP infection or the presence of TLR4 and IL-8 gene polymorphisms and HP infection.

Topics: Adult; Aged; Alleles; Case-Control Studies; Female; Gene Frequency; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Neoplasm Grading; Neoplasm Staging; Odds Ratio; Polymorphism, Genetic; Stomach Neoplasms; Toll-Like Receptor 4

Helicobacter pylori colonization of the gastric epithelium induces interleukin-8 (IL-8) production and inflammation leading to host cell damage. We searched for gastric-derived Lactobacillus with the ability to suppress H. pylori-induced inflammation.. Conditioned media from gastric-derived Lactobacillus spp. were tested for the ability to suppress H. pylori-induced IL-8 production in AGS gastric epithelial cells. IL-8 protein and mRNA levels were measured by ELISA and qPCR, respectively. The changes on host cell signaling pathway were analyzed by Western blotting and the anti-inflammatory effect was tested in a Sprague-Dawley rat model.. Conditioned media from L. salivarius B101, L. rhamnosus B103, and L. plantarum XB7 suppressed IL-8 production and IL-8 mRNA expression in H. pylori-induced AGS cells without inhibiting H. pylori growth. Conditioned media from LS-B101, LR-B103, and LP-XB7 suppressed the activation of NF-κB in AGS cells, while strain LP-XB7 also suppressed c-Jun activation. The anti-inflammatory effect of LP-XB7 was further assessed in vivo using a H. pylori-infected Sprague-Dawley rat model. Strain LP-XB7 contributed to a delay in the detection and colonization of H. pylori in rat stomachs, attenuated gastric inflammation, and ameliorated gastric histopathology. Additionally, the administration of LP-XB7 correlated with the suppression of TNF-α and CINC-1 in sera, and suppression of CINC-1 in the gastric mucosa of H. pylori-infected rats.. These results suggest that L. plantarum XB7 produces secreted factors capable of modulating inflammation during H. pylori infection, and this probiotic Lactobacillus strain shows promise as an adjunctive therapy for treating H. pylori-associated disease.

Topics: Adult; Aged; Animals; Anti-Inflammatory Agents; Culture Media, Conditioned; Disease Models, Animal; Epithelial Cells; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunomodulation; Inflammation; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lactobacillus plantarum; Male; Middle Aged; Probiotics; Rats; Rats, Sprague-Dawley; RNA, Messenger; Signal Transduction; Stomach; Transcription Factor RelA

Helicobacter pylori is a human gastric pathogen that adheres to host cells and injects cytotoxin-associated gene A (CagA) to induce interleukin-8 (IL-8), inducible nitric oxide (iNOS), and cyclooxygenase 2 (COX-2). Noni (Morinda citrifolia) is found to possess antibacteria, anti-inflammation, and antioxidation activities, but its effect on H. pylori infection is still unknown. Ethanol and ethyl acetate extracts of noni fruit were used in this study. The inhibitory effect on CagA and H. pylori-induced IL-8, iNOS, and COX-2 were determined. The coculture medium was collected for measuring neutrophil chemotaxis. Both extracts of noni fruit showed weak inhibition on H. pylori. Both ethanol and ethyl acetate extracts provided antiadhesion of H. pylori to AGS cells and down-regulation on the CagA, IL-8, COX-2, and iNOS expressions. Results also indicated both extracts relieved neutrophil chemotaxis. Noni fruit extracts down-regulated inflammatory responses during H. pylori infection, and the phenolic compounds play key role in antiadhesion.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents; Bacterial Adhesion; Cell Line; Cyclooxygenase 2; Fruit; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Morinda

To study the effect of Weifuchun on inflammation of Helicobacter pylori (Hp)-infected gastric epithelial cells (GES-1) and its correlation with NF-kappaB signaling pathway.. Hp standard home-made strain (CagA +, VacA +) NCTCI 1637 infected GES-1 cells were used. Weifuchun was used as intervention. Weifuchun of different concentrations (5,10, and 20 microg/mL) were screened by MTT assay. A blank group and the model group were set up. Then the growth inhibition rate of drugs on gastric epithelial GES-1 cells was detected with MTT assay. Cell cycle was detected using flow cytometry. The supernatant liquid was separated to detect the contents of IL-8 and IL-4 by ELISA.The protein expression level of NF-kappaB was detected by Western blot analysis.. MTT assay indicated significantly inhibitory effect of Weifuchun on GES-1 cells [5% inhibiting concentration (IC5)] was 10 microg/ml in the Weifuchun group. After GES-1 and Hp were cultured together,the contents of IL-8 in the supernatant were more obviously higher in the model group than in the blank group (P < 0.05), and then gradually decreased. After treatment with different concentrations of Weifuchun, the levels of IL-8 in the supernatant were less when compared with the model group at 12, 24, 48, and 72 h (P < 0.05). The decrement was the most significant in the high dose Weifuchun group. The IL-4 level in the supernatant was obviously lower in the model group than in the blank group. It obviously increased in the high concentration Weifuchun group (P < 0.05). There was no statistical difference in the IL-4 level between middle, low concentration Weifuchun group and the blank group (P > 0.05). The protein expression of intranuclear P65 increased and that of IkBalpha decreased 60 min after Hp infection. But the protein expression of intranuclear P65 decreased and the protein expression of IkBalpha increased after intervention of Weifuchun.. Weifuchun adjusted H. pylori induced IL-8 and IL-4 production by gastric epithelial cells through blocking NF-kappaB pathways. Its mechanisms might possibly lie in inhibiting p65 from entry into nucleus and the degradation of IkBalpha. Weifuchun was an effective drug for treatment of Hp correlated chronic gastritis.

Topics: Cell Line; Drugs, Chinese Herbal; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Proteins; Inflammation; Interleukin-4; Interleukin-8; NF-kappa B; NF-KappaB Inhibitor alpha; Signal Transduction; Transcription Factor RelA

Helicobacter pylori (H. pylori) bacteria are human pathogens causing symptomatic gastritis, peptic ulcer or gastric cancer. Little is known about the kinetics of immune responses in H. pylori infected patients because the initial moment of infection has not been identified. Various animal models are used to investigate the immune processes related to H. pylori infection. In this study we checked whether H. pylori infection in guinea pigs, mimicking natural H. pylori infection in humans, resulted in the development of specific immune responses to H. pylori antigens by measuring the proliferation of lymphocytes localized in mesenteric lymph nodes, spleen and peripheral blood. The maturity of macrophages and cytokines, delivered by monocyte-macrophage lineage or lymphocytes, were considered as mediators, which might influence the lymphocyte blastogenic response. The obtained results showed the activation of T cells localized in mesenteric lymph nodes by H. pylori antigens in H. pylori infected guinea pigs four weeks postinfection. The blastogenic activity of lymphocytes was shaped by their interaction with antigen presenting cells, which were present in the cell cultures during the whole culture period. Moreover, the balance between cytokines derived from adherent leukocytes including interleukin 8--IL-8 as well as interferon gamma--IFN-γ, and transforming growth factor beta--TGF-β delivered by lymphocytes, was probably important for the successful proliferation of lymphocytes. The H. pylori specific lymphocytes were not propagated in peripheral blood and spleen of H. pylori infected animals. The modulation of immunocompetent cells by H. pylori antigens or their different distribution cannot be excluded.

Topics: Animals; Antigens, Bacterial; Cell Proliferation; Cells, Cultured; Dendritic Cells; Guinea Pigs; Helicobacter Infections; Helicobacter pylori; Immunity, Cellular; Interferon-gamma; Interleukin-8; Lymph Nodes; Lymphocyte Activation; Lymphocyte Count; Lymphocytes; Macrophages; Male; Mesentery; Spleen; Transforming Growth Factor beta

Helicobacter pylori (H. pylori) infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer. Total alkaloids of sophora alopecuroides (TASA) is widely used in herbal remedies to treat various infectious diseases, including stomach-associated diseases. This study is aimed at evaluating the antimicrobial activity of TASA on H. pylori-infected BALB/c mice mouse gastritis.. Totally 120 BALB/c mice were orally inoculated with H. pylori Bacterial liquid to construct BALB/c mice H. pylori infection gastritis animal model, after the model was successfully created. We randomly assigned 100 infected mice into 10 treatment groups, the first group (normal saline); the second group (bismuth pectin); the third group (omeprazole); the fourth group (TASA 2 mg/d); the fifth group (TASA 4 mg/d); the sixth group (TASA 5 mg/d); the seventh group (TASA + bismuth pectin); the eighth group (TASA + omeprazole); the ninth group (bismuth pectin + clarithromycin + metronidazole); the tenth group (omeprazole + clarithromycin + metronidazole), 5 other non-infected mice as negative control. Mice were orally inoculated twice a day and 7 days continuously. Then the mice were killed 4 weeks after treatment, we used realtime PCR to detect 16sDNA of H. pylori to test both the colonization and the clearance mice of bacteria of each treatment. We applied hematoxylin and eosin (HE) staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 (IL-8), cyclooxygenase 2 (COX-2), and nuclear factor-kappa B (NF-κB) expression change after treatments.. Firstly, we ensured that after 6-week intragastric administration, the bacteria colonization reached an exceed peak which is far higher than positive threshold (P < 0.001); secondly, after treatments, it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H. pylori as well as conventional triple therapy (P < 0.001); thirdly, HE staining showed that the inflammation on mice gastric mucosal membrane were also relieved obviously in TASA combined treatments and conventional triple therapy compared with normal saline treated mice, moreover, from immunohistochemistry results, H. pylori-induced IL-8, COX-2, and NF-κB were consistently suppressed in seventh, eighth, ninth, and tenth group to a certain extent.. These results open the possibility of taking TASA as an anti-inflammatory agent for H. pylori gastritis.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Cyclooxygenase 2; Female; Helicobacter Infections; Helicobacter pylori; Immunohistochemistry; Interleukin-8; Mice; Mice, Inbred BALB C; NF-kappa B; Omeprazole; Real-Time Polymerase Chain Reaction; Sophora

Outer membrane proteins (OMPs) represent an important class of proteins that are observed in gram-negative bacteria, mitochondria and chloroplasts. These proteins play diverse biological roles in protein translocation, cell-cell communication and signal transduction. A variety of OMPs have been identified in the gastrointestinal pathogen Helicobacter pylori (H. pylori) since it was first isolated in 1983. Among these proteins, outer membrane inflammatory protein A (OipA), which is encoded by hopH and unique to this pathogen, is a differentially expressed outer membrane protein that has been confirmed to be directly linked to H. pylori colonization, as well as to the pathogenesis of H. pylori and disease outcome. In this review, we will describe the progress of recent studies on OipA, particularly those on the functions and biological significance of this unique protein.

Topics: Bacterial Outer Membrane Proteins; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Polymorphism, Genetic; Prognosis; Virulence Factors

Helicobacter pylori is among the major pathogenic bacteria that cause chronic gastritis and peptic ulcer disease and is related to the development of gastric cancer. Several chemicals, including antibiotics, have been used to eradicate H. pylori; however, they do not always curb the infection. Ten representative type strains of lactic acid bacteria (LAB) were screened for antagonism toward H. pylori via inhibition of urease activity. Strains inhibiting the binding of H. pylori to human gastric cell line cells and suppressing H. pylori-induced interleukin-8 (IL-8) production were also screened. Of these, Pediococcus pentosaseus (SL4), which inhibited the adhesion of H. pylori to MKN-45 gastric cancer cells, Bifidobacterium longum (BG7), with urease inhibiting activity, and Lactococcus lactis (SL3), and Enterococcus faecalis (SL5), which suppressed H. pylori-induced IL-8 production within MKN-45 and AGS cells, were selected. In mouse model, these LAB stains in combination significantly suppressed IL-8 levels in serum. Gastric pH also recovered to normal values after the administration of these LAB. These stains effectively suppressed H. pylori viability, although not to the extent of antibiotic treatment. When used as probiotics, LAB may help decrease the occurrence of gastritis and reduce the risk of H. pylori infection without, inducing side effects.

Topics: Animals; Antibiosis; Bacterial Adhesion; Bifidobacterium; Cell Line, Tumor; Gastric Acid; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillales; Male; Mice; Mice, Inbred C57BL; Microbial Viability; Probiotics; Urease

The infection of Helicobacter pylori (H. pylori) is one of the most important causes of gastric ulcer disease. The role of hydrogen sulfide (H2S) production in H. pylori-induced gastric ulcer disease.. The expression of cystathionine-γ-lyase (CSE) was determined, and correlated with the severity of gastric ulcer disease.. One hundred and eight patients were selected based on the determination of gastric ulcer and the infection of Helicobacter pylori (H. pylori), including 36 normal control, 36 patients with H. Pylori-negative gastric ulcer, and 36 patients with H. Pylori-positive gastric ulcer. RT-PCR determination was performed to determine the expression of CSE, NF-κB and IL-8.. The expression of CSE, NF-κB and IL-8 was higher in the gastric ulcer group than control group (p<0.05). Compared with the H. pylori-negative gastric ulcer, the expression of CSE, NF-κB and IL-8 was higher than H. pylori-positive gastric ulcer group (p<0.05). For H. pylori-negative gastric ulcer group, the expression of CSE positively correlated with the expression of NF-κB (r=0.98, p<0.05) and IL-8 (r=0.95, p<0.05). For H. pylori-positive gastric ulcer group, the expression of CSE also positively correlated with the expression of NF-κB (r=0.99, p<0.05) and IL-8 (r=0.85, p<0.05).. The expression of CSE was positively correlated with the severity of gastric ulcer.

Topics: Adult; Case-Control Studies; Cystathionine gamma-Lyase; Female; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen Sulfide; Interleukin-8; Male; Middle Aged; NF-kappa B; Peptic Ulcer; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Severity of Illness Index

Synthetic studies of lipid A and LPS partial structures have been performed to investigate the relationship between structures and functions of LPS. Recent studies have suggested several pathological implications of LPS from parasitic bacteria due to its influence on the host immune responses. To address this issue, we established an efficient synthetic strategy that is widely applicable to the synthesis of various lipid As by using a key disaccharide intermediate with selectively cleavable protecting groups. Porphyromonas gingivalis and Helicobacter pylori lipid As were synthesized and their biological activities were evaluated. All synthetic lipid As did not induce strong inflammatory responses: some are very weak cytokine inducers and others are antagonistic in IL-6 and IL-8 induction with E. coli LPS. On the other hand, P. gingivalis lipid As showed potent IL-18 inducing activity. Since IL-18 has been shown to correlate with chronic inflammation, P. gingivalis LPS may be implicated in the chronic inflammatory responses.

Topics: Bacteroidaceae Infections; Chemistry Techniques, Synthetic; Enzyme-Linked Immunosorbent Assay; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Innate; Immunomodulation; Interleukin-6; Interleukin-8; Lipid A; Lipopolysaccharides; Models, Chemical; Molecular Structure; Porphyromonas gingivalis

Helicobacter pylori causes clinical disease primarily in those individuals infected with a strain that carries the cytotoxin associated gene pathogenicity island (cagPAI). The cagPAI encodes a type IV secretion system (T4SS) that injects the CagA oncoprotein into epithelial cells and is required for induction of the pro-inflammatory cytokine, interleukin-8 (IL-8). CagY is an essential component of the H. pylori T4SS that has an unusual sequence structure, in which an extraordinary number of direct DNA repeats is predicted to cause rearrangements that invariably yield in-frame insertions or deletions. Here we demonstrate in murine and non-human primate models that immune-driven host selection of rearrangements in CagY is sufficient to cause gain or loss of function in the H. pylori T4SS. We propose that CagY functions as a sort of molecular switch or perhaps a rheostat that alters the function of the T4SS and "tunes" the host inflammatory response so as to maximize persistent infection.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Bacterial Secretion Systems; DNA, Bacterial; Female; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Interleukin-8; Macaca mulatta; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Microscopy, Electron, Scanning; Recombination, Genetic; Specific Pathogen-Free Organisms; Virulence Factors

To investigate whether the phenyl-thiophenyl propenone RK-I-123 suppresses interleukin-8 (IL-8) expression and activation of mitogen-activated protein kinases (MAPKs) and transcription factors (nuclear factor-κB [NF-κB] and activator protein-1 [AP-1]) by reducing reactive oxygen species (ROS) levels in Helicobacter pylori-infected gastric epithelial cells.. Helicobacter pylori in Korean isolates, human gastric epithelial AGS cells.. AGS cells pretreated with or without RK-I-123 were cultured in the presence of H. pylori at a bacterium/cell ratio of 300:1.. Reactive oxygen species and IL-8 levels were determined by dichlorofluorescein fluorescence and enzyme-linked immunosorbent assay. The IL-8 mRNA expression was analyzed by the real-time reverse transcription-polymerase chain reaction (RT-PCR). The MAPK and IκBα levels were determined by western blotting. The activation of NF-κB and AP-1 was determined by the electrophoretic mobility shift assay.. Helicobacter pylori induced an increase in ROS and IL-8 expression and activation of MAPKs and transcription factors (NF-κB and AP-1) together with the degradation of IκBα in AGS cells, all of which were inhibited by RK-I-123.. The RK-I-123 suppressed the H. pylori-induced IL-8 expression and activation of MAPKs, NF-κB, and AP-1 by reducing ROS levels in AGS cells. The RK-I-123 may be a potential candidate for the treatment of H. pylori-induced gastric inflammation.

Topics: Anti-Inflammatory Agents; Cells, Cultured; Chalcones; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Reactive Oxygen Species; Thiophenes; Transcription Factor AP-1

While Helicobacter pylori infects over 50% of the world's population, the mechanisms involved in the development of gastric disease are not fully understood. Bacterial, host, and environmental factors play a role in disease outcome. To investigate the role of bacterial factors in H. pylori pathogenesis, global gene expression of six H. pylori isolates was analyzed during coculture with gastric epithelial cells. Clustering analysis of six Colombian clinical isolates from a region with low gastric cancer risk and a region with high gastric cancer risk segregated strains based on their phylogeographic origin. One hundred forty-six genes had increased expression in European strains, while 350 genes had increased expression in African strains. Differential expression was observed in genes associated with motility, pathogenicity, and other adaptations to the host environment. European strains had greater expression of the virulence factors cagA, vacA, and babB and were associated with increased gastric histologic lesions in patients. In AGS cells, European strains promoted significantly higher interleukin-8 (IL-8) expression than did African strains. African strains significantly induced apoptosis, whereas only one European strain significantly induced apoptosis. Our data suggest that gene expression profiles of clinical isolates can discriminate strains by phylogeographic origin and that these profiles are associated with changes in expression of the proinflammatory and protumorigenic cytokine IL-8 and levels of apoptosis in host epithelial cells. These findings support the hypothesis that bacterial factors determined by the phylogeographic origin of H. pylori strains may promote increased gastric disease.

Topics: Adaptation, Physiological; Antigens, Bacterial; Apoptosis; Bacterial Proteins; Cell Line, Tumor; Cluster Analysis; Coculture Techniques; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation, Bacterial; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; Middle Aged; Movement; Phylogeography; Transcriptome

To study gastric mucosal interleukine-8 (IL-8) mRNA expression, the cytotoxin-associated gene A (cagA) mutation, and serum pepsinogen (PG) I/II ratio related risk in Thai gastric cancer.. There were consent 134 Thai non-cancer volunteers who underwent endoscopic narrow band imaging examination, and 86 Thais advance gastric cancer patients who underwent endoscopic mucosal biopsies and gastric surgery. Tissue samples were taken by endoscopy with 3 points biopsies. The serum PG I, II, and Helicobacter pylori (H. pylori) immunoglobulin G (IgG) antibody for H. pylori were tested by enzyme-linked immunosorbent assay technique. The histopathology description of gastric cancer and non-cancer with H. pylori detection was defined with modified Sydney Score System. Gastric mucosal tissue H. pylori DNA was extracted and genotyped for cagA mutation. Tissue IL-8 and cyclooxygenase-2 (COX-2) mRNA expression were conducted by real time relative quantitation polymerase chain reaction. From 17 Japanese advance gastric cancer and 12 benign gastric tissue samples, all were tested for genetic expression with same methods as well as Thai gastric mucosal tissue samples. The multivariate analysis was used for the risk study. Correlation and standardized t-test were done for quantitative data, P value < 0.05 was considered as a statistically significant.. There is a high non cagA gene of 86.8 per cent in Thai gastric cancer although there are high yields of the East Asian type in the positive cagA. The H. pylori infection prevalence in this study is reported by combined histopathology and H. pylori IgG antibody test with 77.1% and 97.4% of sensitivity and specificity, respectively. The serum PG I/II ratio in gastric cancer is significantly lower than in the non-cancer group, P = 0.045. The serum PG I/II ratio of less than 3.0 and IL-8 mRNA expression ≥ 100 or log10 ≥ 2 are significant cut off risk differences between Thai cancer and non-cancer, P = 0.03 and P < 0.001, respectively. There is a significantly lower PGI/II ratio in Japanese than that in Thai gastric cancer, P = 0.026. Serum PG I/II ratio at cut off less than 3.0 and IL-8 mRNA expression Raw RQ > 100 or log10 > 2 are significantly difference between Thai cancer group when compared to non-cancer group, P = 0.013 and P < 0.001, respectively. In the correlation study, low PG I/II ratio does not associate with chronic atrophic gastritis severity score in Thais non-cancer cases. However, there is a trend, but not significant convert correlation between IL-8 mRNA expression level and low PG I/II ratio in Thai positive H. pylori infection. The high expression of IL-8 gene demonstrates a poorer prognosis by stage and histology.. Predominant gastric mucosal IL-8 mRNA expression level, H. pylori infection, and low PG I/II ratio are relative risks for Thai gastric cancer without correlation with cagA mutation.

Topics: Adult; Antigens, Bacterial; Asian People; Bacterial Proteins; Biomarkers; Chi-Square Distribution; Cross-Sectional Studies; Cyclooxygenase 2; Female; Gastric Mucosa; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Multivariate Analysis; Mutation; Odds Ratio; Pepsinogen A; Pepsinogen C; Phenotype; Risk Factors; RNA, Messenger; Stomach Neoplasms; Thailand

Although Helicobacter pylori (Hp) plays an important role in the pathogenesis of chronic gastritis and gastric ulcer, little is known about the probable mechanisms of these types of gastrointestinal damage. To determine the precise mechanisms involved in ulcer formation, immune responses in patients with gastric ulcer (GUP) caused by Hp infection (Hp(+)) were compared with those of other gastritis patients (GP). The sensitivity and proliferation of peripheral blood mononuclear cells (PBMNCs) obtained from patients were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against exposure with complex Hp crude antigen (HPCA) and mitogen (phytohemagglutinin, PHA). Production of inflammatory cytokines, including interleukin (IL)-1β and IL-8, in serum and supernatants of PBMNCs were then measured by ELISA. It was found that, after stimulation with PHA, both IL-8 and IL-1β concentrations in sera and supernatants as well as proliferation and sensitivity were statistically greater in GUP Hp(+) than GP Hp(-) . Furthermore, HPCA inhibited the proliferation of PBMNCs dose-dependently; however, it stimulated IL-8 and IL-1β production in supernatants of mononuclear cells. Therefore, the up-regulated concentrations of IL-8 and IL-1β may have been caused by increase in the size of mononuclear cell subpopulations or in their cytokine secretory activity, indicating the greatest cell responsiveness in GUP Hp(+) patients. These results suggest that tissue damage and ulcers occur in patients who produce more IL-8 and IL-1β than patients who do not develop ulcers; the former consequently have more activated immune cells at the site of infection. Therefore, both host responses and Hp virulence factors may be involved in the development of gastric ulcers.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cell Proliferation; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Staining and Labeling; Stomach Ulcer; Tetrazolium Salts; Thiazoles; Young Adult

The natural course of Helicobacter pylori infection, as well as the success of antibiotic eradication is determined by the immune response to bacteria. The aim of the study is to investigate how different Helicobacter pylori isolates influence the dendritic cells maturation and antigen-presenting function in order to elucidate the differences between Helicobacter pylori strains, isolated from the patients with successful antibiotic eradication therapy or repeated eradication failure.. Dendritic cells maturation and antigen presentation were monitored by flow cytometry analysis of the major histocompatibility complex class II (MHC-II), Toll-like receptor (TLR) and costimulatory molecules expression, and by determining cytokine secretion.. Dendritic cells stimulated with Helicobacter pylori isolated from patients with repeated antibiotic eradication failure expressed less human leukocyte antigen (HLA-DR), CD86, TLR-2, and interleukin-8 (IL-8) compared to Helicobacter pylori strains susceptible to antibiotic therapy; the latter expressed lower production of IL-10. Polymyxin B inhibition of lipopolysaccharide reduces IL-8 secretion in the group of Helicobacter pylori strains susceptible to antibiotic therapy. The differences in IL-8 secretion between both groups are lipopolysaccharide dependent, while the differences in secretion of IL-10 remain unchanged after lipopolysaccharide inhibition. Inhibitor of cathepsin X Mab 2F12 reduced the secretion of IL-6, and the secretion was significantly lower in the group of Helicobacter pylori strains isolated from patients with repeated antibiotic eradication failure.. Helicobacter pylori strains, susceptible/resistant to antibiotic eradication therapy, differ in their capability to induce DCs maturation and antigen-presenting function.

Topics: Anti-Bacterial Agents; Dendritic Cells; Drug Resistance, Bacterial; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interferon-gamma; Interleukin-10; Interleukin-6; Interleukin-8; Male; Toll-Like Receptors

γ-Glutamyltranspeptidase and asparaginase have been shown to play important roles in Helicobacter pylori colonization and cell death induced by H. pylori infection. In this study, the association of γ-glutamyltranspeptidase and asparaginase was elucidated by comparing activities of both deamidases in H. pylori strains from patients with chronic gastritis, gastric and duodenal ulcers, and gastric cancer. γ-Glutamyltranspeptidase activities in H. pylori strains from patients with gastric cancer were significantly higher than in those from patients with chronic gastritis or gastric ulcers. There was a wide range of asparaginase activities in H. pylori strains from patients with gastric cancer and these were not significantly than those from patients with other diseases. To identify the contributions of γ-glutamyltranspeptidase and asparaginase to gastric cell inflammation, human gastric epithelial cells (AGS line) were infected with H. pylori wild-type and knockout strains and inflammatory responses evaluated by induction of interleukin-8 (IL-8). IL-8 response was significantly decreased by knockout of the γ-glutamyltranspeptidase-encoding gene but not by knockout of the asparaginase-encoding gene. Additionally, IL-8 induction by infection with the H. pylori wild-type strain was significantly decreased by adding glutamine during infection. These findings indicate that IL-8 induction caused by γ-glutamyltranspeptidase activity in H. pylori is mainly attributable to depletion of glutamine. These data suggest that γ-glutamyltranspeptidase plays a significant role in the chronic inflammation caused by H. pylori infection.

Topics: Adult; Aged; Aged, 80 and over; Asparaginase; Bacterial Proteins; Base Sequence; Cell Line; Epithelial Cells; gamma-Glutamyltransferase; Gastritis; Gene Knockout Techniques; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Peptic Ulcer; Stomach Neoplasms; Young Adult

H. pylori CagL amino acid polymorphisms such as Y58/E59 can increase integrin α5β1 expression and gastric cancer risk. Hypochlorhydria during chronic H. pylori infection promotes gastric carcinogenesis. The study test whether CagL-Y58/E59 isolates may regulate integrin α5β1 to translocate CagA via the type IV secretory system even under adverse pH conditions, and whether the integrin α5β1 expression primed by H. pylori is a pH-dependent process involving hypochlorhydria in a vicious cycle to promote gastric carcinogenesis.. The expressions of integrin α5 and β1, CagA phosphorylation, IL-8, FAK, EGFR, and AKT activation of AGS cells exposed to CagL-Y58/E59 H. pylori, isogenic mutants, and different H. pylori CagL amino acid replacement mutants under different pH values were determined. Differences in the pepsinogen I/II ratio (indirectly indicating gastric acidity) and gastric integrin α5β1 expression were compared among the 172 H. pylori-infected patients with different cancer risks.. Even under adversely low pH condition, H. pylori CagL-Y58/E59 still keep active integrin β1 with stronger binding affinity, CagA translocation, IL-8, FAK, EGFR, and AKT activation than the other mutants (p<0.05). The in vitro assay revealed higher priming of integrin α5β1 by H. pylori under elevated pH as hypochlorhydria (p<0.05). In the H. pylori-infected patients, the gastric integrin α5β1 expressions were higher in those with pepsinogen I/II ratio <6 than in those without (p<0.05).. H. pylori CagL-Y58/E59 prime higher integrin under adverse pH and may involve to enhance hypochlorhydria vicious cycle for gastric carcinogenesis, and thus require an early eradication.

Topics: Achlorhydria; Adult; Aged; Bacterial Proteins; Cell Transformation, Neoplastic; ErbB Receptors; Female; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Integrin alpha5beta1; Interleukin-8; Male; Middle Aged; Models, Biological; Mutation; Pepsinogen A; Pepsinogen C; Phosphorylation; Protein Binding; Protein Transport; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Stomach Neoplasms

Helicobacter bilis (H. bilis) infection is associated with cases of inflammatory bowel Disease, thyphlocolitis, hepatitis and cholecystitis. However, little is known about the bacterial virulence determinants or the molecular mechanisms involved. Recently, H. bilis γ-glutamyltranspeptidase (HBgGT) was shown to be a virulence factor decreasing host cell viability. Bacterial gGTs play a key role in synthesis and degradation of glutathione and enables the bacteria to utilize extracellular glutamine and glutathione as sources of glutamate. gGT-mediated loss of cell viability has so far been linked to DNA damage via oxidative stress, but the signaling cascades involved herein have not been described. In this study, we identified enhanced ROS production induced by HBgGT as a central factor involved in the activation of the oxidative stress response cascades, which finally activate CREB, AP-1 and NF-κB in H. bilis infected colon cancer cells. IL-8, an important pro-inflammatory chemokine that is a common downstream target of these transcription factors, was up-regulated upon H. bilis infection in an HBgGT dependent manner. Moreover, the induction of these signaling responses and inflammatory cytokine production in host cells could be linked to HBgGT-mediated glutamine deprivation. This study implicates for the first time HBgGT as an important regulator of signaling cascades regulating inflammation in H. bilis infected host epithelial cells that could be responsible for induction of inflammatory disorders by the bacterium.

Topics: Cell Line, Tumor; Colon; Colonic Neoplasms; Cyclic AMP Response Element-Binding Protein; gamma-Glutamyltransferase; Glutamine; Helicobacter; Helicobacter Infections; Humans; Interleukin-8; Intestinal Mucosa; NF-kappa B; Oxidative Stress; Reactive Oxygen Species; Signal Transduction; Transcription Factor AP-1; Transcriptional Activation

Recently, there has been a growing interest in an expanding group of cytokines known as "IL-12 family". The so far gained knowledge about these cytokines, as crucial playmakers in mucosal immunity, has not yet been sufficiently investigated in the context of Helicobacter pylori infection. All genes encoding the monomeric components of these cytokines and their corresponding receptors were examined in gastric epithelial cell lines (AGS and MKN-28) after being infected with 4 H. pylori strains: BCM-300, P1 wild-type, and P1-derived isogenic mutants lacking cytotoxin-associated gene A (cagA) or virulence gene virB7 (multiplicity of infection=50). Both infected and uninfected samples were analyzed after 24h and 48h using real-time quantitative polymerase chain reaction (RT-qPCR). Gene expression analysis demonstrated a strong upregulation of IL23A (encodes p19) by infection, whereas IL23R, Epstein-Barr virus-induced gene 3 (EBI3), IL6ST, IL12A, and IL27RA were found to be expressed, but not regulated, or to a lesser extent. Transcripts of IL12RB2, IL12B, IL12RB1, and IL27A were not detected. Interestingly, P1 resulted in stronger alterations of expression than CagA mutant and BCM-300, particularly for IL23A (59.7-fold versus 32.4- and 6.7-fold, respectively in AGS after 48h, P<.05), whereas no changes were seen with VirB7 mutant. In a proof-of-principle experiment, we demonstrated epithelial-derived expression of IL-12, p19, and Ebi3 in gastric mucosa of gastritis patients using immunohistochemistry (IHC). Unlike IL-12 and Ebi3, increased immunostaining of p19 was observed in H. pylori gastritis. Herein, we highlight the potential role of gastric epithelial cells in mucosal immunity, not only because they are predominant cell type in mucosa and initial site of host-bacterial interaction, but also as a major contributor to molecules that are thought to be primarily expressed by immune cells so far. Of these molecules, p19 was the most relevant one to H. pylori infection in terms of expression and localization.

Topics: Cell Line; Gastric Mucosa; Gastritis; Gene Expression; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-12; Interleukin-23 Subunit p19; Interleukin-8; Interleukins; Minor Histocompatibility Antigens; Multigene Family; Receptors, Interleukin; Transcriptome

To investigate the effect of hydrogen sulfide (H2S) on the expression of CSE, NF-κB, and IL-8 mRNA in GES-1 cells with Helicobacter pylori (H. pylori) infection and to explore its mechanism on gastric mucosa inflammation caused by H. pylori.. GES-1 cells were cultured for 24 h and divided into a control group (neither H. pylori nor NaHS), an H. pylori group, a NaHS group (which was further divided into 4 groups at 50, 100, 200, or 400 μmol/L NaHS), and H. pylori + NaHS group (which was further divided into 4 groups at 50, 100, 200, or 400 μmol/L NaHS). Each group was then cultured for 3, 6, or 12 h. The expression of CSE, NF-κB, and IL-8 mRNA was measured by RT-PCR, and their correlation was analyzed.. The expression of CSE, NF-κB, and IL-8 mRNA in GES-1 cells in the H. pylori group was higher than that in the control group. The expression of CSE in the 200 μmol/L NaHS group and 400 μmol/L NaHS group was lower than that of the control group (P<0.05), whereas the expression of NF-κB and IL-8 in all NaHS groups had no statistical differences compared with the control group (P>0.05). The expression of CSE, NF-κB, and IL-8 mRNA in all groups of NaHS, H. pylori + 200 μmol/L NaHS group, and H. pylori + 400 μmol/L NaHS group was lower than that in the H. pylori group (P<0.05). There was positive correlation among the expressions of CSE, NF-κB, and IL-8 mRNA in the H. pylori group, the H. pylori + 200 μmol/L NaHS group, and the H. pylori + 400 μmol/L NaHS group (P<0.05).. H. pylori can induce NF-κB and IL-8 mRNA expression and upregulate CSE mRNA expression. At 200 and 400 μmol/L, NaHS can suppress H. pylori-induced NF-κB and IL-8 mRNA expression and ameliorate the morphology of H. pylori-induced GES-1 injury, which may protect gastric epithelial cells by H. pylori infection.

Topics: Cell Line; Cystathionine gamma-Lyase; Epithelial Cells; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen Sulfide; Interleukin-8; NF-kappa B; RNA, Messenger; Sulfides

Interleukin (IL)-8 has an important role in initiating inflammation in humans, attracting immune cells such as neutrophils through their receptors CXCR1 and CXCR2. IL-8 has been proposed to contribute to chronic inflammation and cancer. However, mice do not have the IL-8 gene, so human cancer cell lines and xenograft studies have been used to study the role of IL-8 in colon and gastric carcinogenesis. We generated mice that carry a bacterial artificial chromosome that encompasses the entire human IL-8 gene, including its regulatory elements (IL-8Tg mice).. We studied the effects of IL-8 expression in APCmin(+/-) mice and IL-8Tg mice given azoxymethane and dextran sodium sulfate (DSS). We also examined the effects of IL-8 expression in gastric cancer in INS-GAS mice that overexpress gastrin and IL-8Tg mice infected with Helicobacter felis.. In IL-8Tg mice, expression of human IL-8 was controlled by its own regulatory elements, with virtually no messenger RNA or protein detectable under basal conditions. IL-8 was strongly up-regulated on systemic or local inflammatory stimulation, increasing mobilization of immature CD11b(+)Gr-1(+) myeloid cells (IMCs) with thioglycolate-induced peritonitis, DSS-induced colitis, and H. felis-induced gastritis. IL-8 was increased in colorectal tumors from patients and IL-8Tg mice compared with nontumor tissues. IL-8Tg mice developed more tumors than wild-type mice following administration of azoxymethane and DSS. Expression of IL-8 increased tumorigenesis in APCmin(+/-) mice compared with APCmin(+/-) mice that lack IL-8; this was associated with increased numbers of IMCs and angiogenesis in the tumors.. IL-8 contributes to gastrointestinal carcinogenesis by mobilizing IMCs and might be a therapeutic target for gastrointestinal cancers.

Topics: Animals; Azoxymethane; Cell Line, Tumor; Cell Movement; Cell Transformation, Neoplastic; Colitis; Colonic Neoplasms; Dendritic Cells; Dextran Sulfate; Gastritis; Helicobacter felis; Helicobacter Infections; Humans; Interleukin-8; Lipopolysaccharides; Macrophages; Mice; Mice, Transgenic; Myeloid Cells; Primary Cell Culture; RNA, Messenger; Tumor Burden; Up-Regulation

Helicobacter pylori is associated with the majority of gastric disorders and the antibiotic resistant rates have increased annually worldwide. Anisomeles indica and its constituent, ovatodiolide (OVT), were shown to have bactericide activity against Helicobacter pylori. The aim of this study was to manufacture extracts containing the effective constituent, OVT, and evaluate their bactericidal function and the inhibition of inflammatory responses to Helicobacter pylori infection.. Various concentrations of ethanol for extraction of Anisomeles indica were performed and the content of OVT was analyzed by high-performance liquid chromatography (HPLC). The anti-bacterial activity of Anisomeles indica ethanol extracts and the constituent OVT were determined. Additional experiments were performed to investigate the Anisomeles indica ethanol extracts and OVT to inhibit the Helicobacter pylori-induced inflammation of both gastric epithelial cells and macrophages.. Amongst the extracts tested, 50% and 95% ethanol extracts contained large amount of OVT and showed potent anti-Helicobacter pylori activity. An in vitro Helicobacter pylori-infection model revealed that 95% ethanol extract attenuated Helicobacter pylori-induced nuclear factor kappa B (NF-κB) activity and interleukin (IL)-8 secretion of gastric epithelial cells. In addition, 95% ethanol extract significantly inhibited lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS), as well as production of nitric oxide (NO) and tumor necrosis factor α (TNF-α) by macrophages.. This study reveals that Anisomeles indica ethanol extracts containing OVT may be a potent and economic therapeutic agent for Helicobacter pylori infection and attenuation of Helicobacter pylori-mediated inflammation.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Line, Tumor; Cell Survival; Disease Models, Animal; Diterpenes; Epithelial Cells; Ethanol; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Inflammation Mediators; Interleukin-8; Lamiaceae; Macrophages; Mice; Microbial Sensitivity Tests; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Phytotherapy; Plant Extracts; Plant Stems

Studies have demonstrated that some polymorphisms in different interleukin genes may increase the risk of cancer. The aim of this study was to investigate the association between the IL-8 (rs4073) -251A/T gene polymorphism and the risk of gastric cancer (GC).. A case-control study was conducted on patients with noncardia gastric cancer. DNA was extracted from leukocytes and the IL-8 (rs4073) -251A/T polymorphism was analyzed by PCR-RFLP. Infection with Helicobacter pylori was investigated in the serum by ELISA.. The sample consisted of 104 patients with GC and 196 controls. Cigarette smoking (P=0.007) and high fat intake (P=0.01) were more frequent in patients with GC. The proportion of patients infected with H. pylori was similar in the two groups (P=0.101). The frequency of the genotype A/T was higher in the cancer group (P=0.008). An increased risk of GC was found in subjects carrying the genotype A/T (OR=2.50, CI: 1.27-4.90), subjects with high fat intake (OR=1.92, CI: 1.17-3.15), and smokers (OR=2.00, CI: 1.203.31).. Subjects with the heterozygous A/T genotype, high fat intake and smokers or ex-smokers presented an increased risk of GC. Individuals with A/A genotype may have protective effect for GC.

Topics: Brazil; Case-Control Studies; Female; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Kaplan-Meier Estimate; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Risk Factors; Stomach Neoplasms

It has been reported that CagA gene positive Helicobacter pylori (CagA+ H. pylon) induces severe gastric mucosal inflammation. On the other hand, Interleukin (IL)-17 is known to stimulate IL-8 release by the gastric epithelial cells which facilitates chemotaxis of neutrophils through an IL-8-dependent mechanism. The aim of the study is to determine the role of IL-17 and IL-8 in the development of gastritis and gastric ulcer in H. pylori infected patients. Mucosal biopsy samples were obtained from the ulcer site of gastric mucosa of 28 patients with gastric ulcer (GU), 27 with gastritis and 8 controls subjects without gastritis or ulcers. Infection with H. pylori of patients and controls was assessed by a rapid urease test, histological examination and culture. Measurement of the tissue levels of IL-17 and IL-8 were assayed by ELISA. H. pylori cagA gene was assessed by polymerase chain reaction (PCR). Out of the 28 patients with GU, 18 (64.2%) patients were positive for H. pylori infection, while 13 (48.1%) patients with gastritis and none of the controls were positive for H. pylori infection The CagA gene was detected in 12 (66.6%) in H. pylori GU patients, and 7 (53.8%) H. pylori positive gastritis. IL-17 was significantly higher in GU-CagA+ve H. pylori compared to GU-CagA- H. pylori (P <0.05), while IL-8 showed no significant difference between groups. The mean levels of IL-8 in gastritis-CagA+ H. pylori) was significantly higher compared to gastritis--CagA- H. pylori- (P <0.05). IL-17 showed significant association with the number of neutrophils in both GU and gastritis (r = 0.689, P < 0.05 & r = 0.618, P < 0.05). Also, IL-8 showed significant association with the number of neutrophils in both GU and gastritis n (r = 0.468, P < 0.05 & r = 0.727, P < 0.05). It is concluded that the Cag+ve H. pylori is associated with induction of mucosal injury. Also, IL-8 and IL-17 plays a role in the development of GU and gastritis especially in CagA+ H. pylori.

Topics: Antigens, Bacterial; Bacterial Proteins; Enzyme-Linked Immunosorbent Assay; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-7; Interleukin-8; Polymerase Chain Reaction; Stomach Ulcer

The duodenal ulcer promoting (dupA) gene, located in the plasticity region of Helicobacter pylori, is associated with duodenal ulcer development. dupA was predicted to form a type IV secretory system (T4SS) with vir genes around dupA (dupA cluster). We investigated the prevalence of dupA and dupA clusters and clarified associations between the dupA cluster status and clinical outcomes in the U.S. population. In all, 245 H. pylori strains were examined using PCR to evaluate the status of dupA and the adjacent vir genes predicted to form T4SS, in addition to the status of cag pathogenicity island (PAI). The associations between dupA cluster status and interleukin-8 (IL-8) and IL-12 production were also examined. The presence of dupA and all adjacent vir genes were defined as a complete dupA cluster. Many variations related to the status of dupA and dupA cluster genes were identified. Concurrent H. pylori infection and the presence of a complete dupA cluster increases duodenal ulcer risk compared to H. pylori infection with incomplete dupA cluster or without the dupA gene independent on the cag PAI status (adjusted odds ratio, 2.13; 95% confidence interval, 1.13 to 4.03). Gastric mucosal IL-8 levels were also significantly higher in the complete dupA cluster group than in other groups (P=0.01). In conclusion, although the causal relationship between the dupA cluster and duodenal ulcer development is not proved, the presence of a complete dupA cluster but not dupA alone, is associated with duodenal ulcer development.

Topics: Adult; Aged; DNA, Bacterial; Duodenal Ulcer; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Multigene Family; Polymerase Chain Reaction; United States; Virulence; Virulence Factors

To investigate genetic diversity of Helicobacter pylori (H. pylori) cell division-related gene A (cdrA) and its effect on the host response.. Inactivation of H. pylori cdrA, which is involved in cell division and morphological elongation, has a role in chronic persistent infections. Genetic property of H. pylori cdrA was evaluated using polymerase chain reaction and sequencing in 128 (77 American and 51 Japanese) clinical isolates obtained from 48 and 51 patients, respectively. Enzyme-linked immunosorbent assay was performed to measure interleukin-8 (IL-8) secretion with gastric biopsy specimens obtained from American patients colonized with cdrA-positive or -negative strains and AGS cells co-cultured with wild-type HPK5 (cdrA-positive) or its derivative HPKT510 (cdrA-disruptant). Furthermore, the cytotoxin-associated gene A (cagA) status (translocation and phosphorylation) and kinetics of transcription factors [nuclear factor-kappa B (NF-κB) and inhibition kappa B] were investigated in AGS cells co-cultured with HPK5, HPKT510 and its derivative HPK5CA (cagA-disruptant) by western blotting analysis with immunoprecipitation.. Genetic diversity of the H. pylori cdrA gene demonstrated that the cdrA status segregated into two categories including four allele types, cdrA-positive (allele types;Iand II) and cdrA-negative (allele types; III and IV) categories, respectively. Almost all Japanese isolates were cdrA-positive (I: 7.8% and II: 90.2%), whereas 16.9% of American isolates were cdrA-positive (II) and 83.1% were cdrA-negative (III: 37.7% and IV: 45.5%), indicating extended diversity of cdrA in individual American isolates. Comparison of each isolate from different regions (antrum and corpus) in the stomach of 29 Americans revealed that cdrA status was identical in both isolates from different regions in 17 cases. However, 12 cases had a different cdrA allele and 6 of them exhibited a different cdrA category between two regions in the stomach. Furthermore, in 5 of the 6 cases possessing a different cdrA category, cdrA-negative isolate existed in the corpus, suggesting that cdrA-negative strain is more adaptable to colonization in the corpus. IL-8 secretions from AGS revealed that IL-8 levels induced by a cdrA-disrupted HPKT510 was significantly lower (P < 0.01) compared to wild-type HPK5: corresponding to 50%-60% of those of wild-type HPK5. These data coincided with in vivo data that an average value of IL-8 in biopsy specimens from cdrA-positive and cdrA-negative groups was 215.6 and 135.9 pg/mL, respectively. Western blotting analysis documented that HPKT510 had no effect on CagA translocation and phosphorylation, however, nuclear accumulation of NF-κB was lower by HPKT510 compared to HPK5.. Colonization by a cdrA-negative or cdrA-dysfunctional strain resulted in decreased IL-8 production and repression of NF-κB, and hence, attenuate the host immunity leading to persistent infection.

Topics: Bacterial Proteins; Cell Cycle Proteins; Cell Line; Genetic Variation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B

Inflammation is widely recognized as a risk factor for gastric H. pylori-associated disease and disruption of this process provides a potential target for intervention. Using an in vitro system, broccoli sprouts, manuka honey and omega-3 oil, singly and in combination, were screened for their ability to limit H. pylori-associated inflammation. Each food significantly attenuated the release of IL-8 by H. pylori-infected cells, although the magnitude of this effect was variable. Only broccoli sprouts (0.125 mg/mL, w/v) were able to inhibit IL-8 release in response to TNFα, suggesting it acted by a different mechanism to the other two foods. The combination of manuka honey (1.25%, v/v) with omega-3 oil (0.006%, v/v) failed further to reduce IL-8 levels below those observed with honey alone, but the same concentrations of omega-3 oil and manuka honey independently enhanced the antiinflammatory effect of the isothiocyanate-rich broccoli sprouts. The results suggest that in the future certain foods may find increased clinical use as a non-antimicrobial approach for reducing the inflammation that is a major risk factor for H. pylori-associated disease, notably gastric cancer.

Topics: Brassica; Cell Line; Fatty Acids, Omega-3; Functional Food; Helicobacter Infections; Helicobacter pylori; Honey; Humans; Inflammation; Interleukin-8; Tumor Necrosis Factor-alpha

H. pylori infection may trigger Smad7 and NFκB expression in the stomach, whereas probiotics promote gastrointestinal health and improve intestinal inflammation caused by pathogens. This study examines if probiotics can improve H. pylori-induced gastric inflammation by inactivating the Smad7 and NFκB pathways.. Challenge with H. pylori increased IL-8 and TNF-α expressions but not TGF-β1 in MKN45 cells. The RNA levels of Smad7 in AGS cells increased after H. pylori infection in a dose-dependent manner. A higher dose (MOI 100) of L. acidophilus pre-treatment attenuated the H. pylori-induced IL-8 expressions, but not TGF-β1. Such anti-inflammatory effect was mediated via increased cytoplasmic IκBα and depletion of nuclear NFκB. L. acidophilus also inhibited H. pylori-induced Smad7 transcription by inactivating the Jak1 and Stat1 pathways, which might activate the TGF-β1/Smad pathway. L. acidophilus pre-treatment ameliorated IFN-γ-induced Smad7 translation level and subsequently reduced nuclear NF-κB production, as detected by western blotting.. H. pylori infection induces Smad7, NFκB, IL-8, and TNF-α production in vitro. Higher doses of L. acidophilus pre-treatment reduce H. pylori-induced inflammation through the inactivation of the Smad7 and NFκB pathways.

Topics: Cell Line, Tumor; Cell Survival; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Proteins; Inflammation; Interleukin-8; Lactobacillus acidophilus; NF-kappa B; NF-KappaB Inhibitor alpha; Probiotics; Signal Transduction; Smad7 Protein; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Helicobacter pylori, the causative agent of type B gastritis, peptic ulcers, gastric adenocarcinoma and MALT lymphoma, uses the Cag type IV secretion system to induce a strong proinflammatory response in the gastric mucosa and to inject its effector protein CagA into gastric cells. CagA translocation results in altered host cell gene expression profiles and cytoskeletal rearrangements, and it is considered as a major bacterial virulence trait. Recently, it has been shown that binding of the type IV secretion apparatus to integrin receptors on target cells is a crucial step in the translocation process. Several bacterial proteins, including the Cag-specific components CagL and CagI, have been involved in this interaction. Here, we have examined the localization and interactions of CagI in the bacterial cell. Since the cagI gene overlaps and is co-transcribed with the cagL gene, the role of CagI for type IV secretion system function has been difficult to assess, and conflicting results have been reported regarding its involvement in the proinflammatory response. Using a marker-free gene deletion approach and genetic complementation, we show now that CagI is an essential component of the Cag type IV secretion apparatus for both CagA translocation and interleukin-8 induction. CagI is distributed over soluble and membrane-associated pools and seems to be partly surface-exposed. Deletion of several genes encoding essential Cag components has an impact on protein levels of CagI and CagL, suggesting that both proteins require partial assembly of the secretion apparatus. Finally, we show by co-immunoprecipitation that CagI and CagL interact with each other. Taken together, our results indicate that CagI and CagL form a functional complex which is formed at a late stage of secretion apparatus assembly.

Topics: Antigens, Bacterial; Bacterial Proteins; Base Sequence; Gastric Mucosa; Gene Deletion; Genetic Complementation Test; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Immunoprecipitation; Integrins; Interleukin-8; Molecular Sequence Data; Operon; Protein Binding; Protein Transport

Interaction of Helicobacter pylori with gastric mucosa leads to marked cellular and humoral host immunologic responses. The signaling pathways initiated by bacteria-host interaction that result in perturbations in cell structure and function remain unclear. Forkhead transcription factors of class O (FoxO) are implicated in the regulation of apoptosis, cell survival, and pathogenesis. H. pylori infection of gastric epithelial cells induces phosphoinositide-3 kinase (PI3K)-dependent Akt activation and cell survival signaling. We investigated the role of H. pylori-activated PI3K/Akt in the regulation of FoxO1/3a in gastric cells.. Immunoblot, immunoprecipitation, and fluorescence microscopy were used to assess the effect of infection of gastric epithelial cells with wild-type H. pylori and their isogenic cag pathogenicity island (PAI) or oipA mutants on the FoxO1/3a signaling pathways. Interleukin-8 release was determined by enzyme-linked immunosorbent assays..  H. pylori infection resulted in activation of the PI3K p85 subunit and inactivation of FoxO1 and FoxO3a by their phosphorylation and translocation of from the nucleus to the cytoplasm. Inhibition of PI3K or Akt kinase activity reduced FoxO1/3a phosphorylation. Akt, FoxO1, or FoxO3a siRNA reduced H. pylori-induced interleukin-8 production. Infection with oipA mutants reduced PI3K/Akt activation and inhibited FoxO1/3a phosphorylation, whereas infection with cag PAI mutants reduced PI3K/Akt activity but did not inhibit FoxO1/3a activation.. FoxO1 and FoxO3a are novel nuclear substrates of H. pylori-induced PI3K/Akt cell survival signaling pathways that partially control interleukin-8 production. OipA-regulated interleukin-8 release through PI3K/Akt is dependent on FoxO1/3a inactivation, whereas cag PAI-mediated interleukin-8 production employs FoxO1/3-independent signaling.

Topics: Cell Line, Tumor; Epithelial Cells; Forkhead Box Protein O1; Forkhead Box Protein O3; Forkhead Transcription Factors; Gastric Mucosa; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction

The human pathogen Helicobacter pylori employs a diverse collection of outer membrane proteins to colonize, persist, and drive disease within the acidic gastric environment. In this study, we sought to elucidate the function of the host-induced gene HP0289, which encodes an uncharacterized outer membrane protein. We first generated an isogenic H. pylori mutant that lacks HP0289 and found that the mutant has a colonization defect in single-strain infections and is greatly outcompeted in mouse coinfection experiments with wild-type H. pylori. Furthermore, we used protease assays and biochemical fractionation coupled with an HP0289-targeted peptide antibody to verify that the HP0289 protein resides in the outer membrane. Our previous findings showed that the HP0289 promoter is upregulated in the mouse stomach, and here we demonstrate that HP0289 expression is induced under acidic conditions in an ArsRS-dependent manner. Finally, we have shown that the HP0289 mutant induces greater expression of the chemokine interleukin-8 (IL-8) and the cytokine tumor necrosis factor alpha (TNF-α) in gastric carcinoma cells (AGS). Similarly, transcription of the IL-8 homolog keratinocyte-derived chemokine (KC) is elevated in murine infections with the HP0289 mutant than in murine infections with wild-type H. pylori. On the basis of this phenotype, we renamed HP0289 ImaA for immunomodulatory autotransporter protein. Our work has revealed that genes induced in vivo play an important role in H. pylori pathogenesis. Specifically, the outer membrane protein ImaA modulates a component of the host inflammatory response, and thus may allow H. pylori to fine tune the host immune response based on ImaA expression.

Topics: Acids; Animals; Bacterial Outer Membrane Proteins; Cell Line; Disease Models, Animal; Epithelial Cells; Gene Deletion; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Immune Evasion; Immunologic Factors; Interleukin-8; Male; Mice; Tumor Necrosis Factor-alpha; Virulence Factors

As a microaerobe, Helicobacter pylori employs the global regulator SpoT for defending against oxidative stress in vitro. However, the mechanisms how SpoT affects bacterial gene expression is still unknown. Moreover, the function of SpoT in H. pylori colonization in the host is remaining undetermined. To explore the functions of the SpoT in H. pylori pathogenesis, we constructed H. pylori 26695 spoT-deficient mutant (ΔspoT). While grown in ambient atmosphere, protein expression profile of the ΔspoT was analyzed with 2D gel electrophoresis and real-time PCR. Compared to the wild type, the spoT-deficient strain downregulated its transcription of the oxidative-induced genes, as well as the genes responsible for protein degradation and that related to energy metabolism. Meanwhile, the colonization ability of ΔspoT strains in Mongolian gerbil was tested, the results demonstrated a decayed colonization in the mouse stomach with ΔspoT than the wild type. As a matter of facts, the AGS cells infected with the ΔspoT strains excreted increased level of the gastric inflammation cytokines IL-8, and the ΔspoT strains showed poor survival ability when treated with reactive oxygen stress (sodium nitroprusside). The elevated capacity of stimulating cytokines and fragility to reactive oxygen stress may be contribute to decreased colonization of the spoT-deficient mutant in the mouse stomach. Conclusively, we speculate that spoT is a key regulator of the genes for H. pylori spreading in the air and colonization in host stomach.

Topics: Aerobiosis; Animals; Bacterial Proteins; Colony Count, Microbial; Culture Media; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Microbial Viability; Mutation; Nitroprusside; Oxidants; Oxidative Stress; Proteomics; Pyrophosphatases; Real-Time Polymerase Chain Reaction; Stomach

Chronic gastritis associated with Helicobacter pylori is a leading cause of gastric intestinal metaplasia (IM), which arises from abnormal cell differentiation of the epithelium in the gastric mucosa. However, the mechanisms involved in H. pylori-mediated IM remain elusive. The aim of our study was to explore the effects and the underlying mechanisms of H. pylori on the abnormal expression of Hath1 and Sox2 and to reveal its relationship to the development of gastric IM. We found that Hath1 and Sox2 were overexpressed in gastric IM tissue. Hath1 expression was up-regulated, whereas Sox2 expression, which was independent of the CagA virulence factor, was down-regulated in gastric epithelial cells and coincided with increased IL-6 and IL-8 levels in the culture media. Stimulation with H. pylori-related cytokine IL-8, but not IL-6 or IL-1β, was induced by Hath1 expression in the gastric epithelial cells. Although IL-8 and IL-6 levels correlated with STAT3 (signal transducer and activator of transcription) phosphorylation before and after H. pylori eradication in the gastric mucosa, only the blocking of IL-8-induced STAT3 activation using AG490 or STAT3-targeting RNA interference altered Hath1 expression. Additionally, we found that H. pylori down-regulated Hes1, which is a direct downstream target gene of Notch signaling and a repressor of Hath1 expression. These findings suggest that H. pylori induced inflammation up-regulate Hath1 expression via interleukin-8/STAT3 (IL-8) phosphorylation while suppressing Hes1, which provides a novel molecular connection between a H. pylori infection and intestinal metaplasia.

Topics: Antigens, Bacterial; Bacterial Proteins; Basic Helix-Loop-Helix Transcription Factors; Cell Line, Tumor; Coculture Techniques; Dose-Response Relationship, Drug; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Homeodomain Proteins; Humans; Immunity, Mucosal; Immunohistochemistry; Interleukin-6; Interleukin-8; Metaplasia; Phosphorylation; Prospective Studies; RNA Interference; STAT3 Transcription Factor; Transcription Factor HES-1; Tyrphostins

Helicobacter pylori (H. pylori) infection generally protects patients from erosive esophagitis through reduction of acid production due to gastric mucosal atrophy. However, there are H. pylori infected patients who still have erosive esophagitis. The reason for this discrepancy remains unclear. We have previously reported that polymorphisms in IL-8 promoter region influence the susceptibility of H. pylori related diseases. On the other hand, nucleotide-binding oligomerization domain 1 (NOD1) is known to play an important role in H. pylori infection. Hence, we hypothesized polymorphisms of these two molecules in H. pylori infected patients may influence the susceptibility to erosive esophagitis. Genomic DNA was extracted from 312 H. pylori infected Japanese, consisting of 110 patients with erosive esophagitis and 202 healthy controls. ND1+32656 T/GG and IL-8-251 A/T polymorphisms were genotyped by direct sequencing. ND1+32656 GG allele and IL-8-251 T/T allele increased the risk of erosive esophagitis with odds ratio (OR) of 1.9 (95% confidence interval (CI) 1.1-3.0, p=0.013) and 1.7 (95% CI 1.1-2.8, p=0.036), respectively. Combination of these two alleles increased the risk with OR of 3.2(95% CI 1.6-6.5, p=0.001). In conclusion, ND1+32656 GG and IL-8-251 T/T allele may be associated with less reactivity to H. pylori infection, and may increase the risk of erosive esophagitis even in H. pylori infected Japanese population.

Topics: Aged; Alleles; Asian People; Atrophy; Case-Control Studies; Esophagitis; Female; Gastric Acid; Gastric Mucosa; Gene Frequency; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Japan; Male; Middle Aged; Nod1 Signaling Adaptor Protein; Polymorphism, Single Nucleotide

The current therapy for Helicobacter pylori infection includes antimicrobial agents and proton pump inhibitors. We have examined the ability of Lactobacillus spp. to inhibit H. pylori infection.. Probiotic strains isolated from samples of adult feces, infant feces, breast milk, and vaginal swab collected from healthy volunteers in Taiwan and commercially available strains were screened for antagonism toward H. pylori. Inhibition liquid culture assay was used to screen potential anti-H. pylori activity. Then, we performed agar plate inhibition assay, and assays to determine the capacity of probiotics for adhesion, and inhibition and killing of H. pylori, and measured the levels of IL-8 and IL-10. Using animal models, we studied regulation of gastric acid and histopathological changes accompanying anti-H. pylori activity.. We found that six of the tested strains suppressed urease activity of H. pylori: Lactobacillus acidophilus TYCA08, L. acidophilus TYCA15, L. johnsonii MH-68, and L. salivarius subsp. salicinius AP-32 were more effective than the others. In vivo, L. johnsonii MH-68 and L. salivarius subsp. salicinius AP-32 alone or in combination, reduced the H. pylori load in the gastric mucosa, and also reduced inflammatory chemokine expression and lymphocyte infiltration.. Lactobacillus johnsonii MH-68 and L. salivarius subsp. salicinius AP-32 effectively suppress H. pylori viability, and when used as probiotics, they may help decrease the occurrence of gastritis, and even reduce the risk of H. pylori infection.

Topics: Adult; Animals; Antibiosis; Bacterial Adhesion; Bacterial Load; Cell Line; Disease Models, Animal; Female; Helicobacter Infections; Helicobacter pylori; Humans; Infant; Interleukin-10; Interleukin-8; Lactobacillus; Male; Mice; Milk, Human; Probiotics; Rats; Rats, Sprague-Dawley; Taiwan; Treatment Outcome; Vagina

Most of the Helicobacter pylori strains containing the cag pathogenicity island (PAI) have been associated with more severe gastric disease in infected humans. The cag PAI is composed of 27 proteins, and some of the components are required for CagA translocation into host cells as well as induction of proinflammatory cytokines, such as interleukin-8 (IL-8); however, the exact function of most of the components remains unknown or poorly characterized. In this study, we demonstrated that CagT (HP0532), which is an essential structural component of the cag PAI apparatus, plays an important role in the translocation of CagA into host epithelial cells. In addition to being located on the bacterial surface, CagT is also partially localized in the inner membrane, where it acts as a chaperone-like protein and promotes CagA translocation. However, CagT secretion was not detected by immunoprecipitation analysis of cell culture supernatants. Meanwhile, CagT was related to the introduction of IL-8 of the host cell. These results suggest that CagT is expressed on both the inner and outer bacterial membranes, where it serves as a unique type IV secretion system component that is involved in CagA secretion and cag PAI apparatus assembly.

Topics: Antigens, Bacterial; Bacterial Proteins; Bacterial Secretion Systems; Bacterial Translocation; Cell Line, Tumor; Cell Membrane; Epithelium; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Immunoprecipitation; Interleukin-8; Molecular Chaperones; Phosphorylation; Plasmids; Protein Interaction Mapping; Protein Transport; Virulence Factors

Previous studies suggested that polymorphisms of proinflammatory cytokine genes are important host genetic factors in Helicobacter pylori infection. The present study evaluated whether IL-8-251 polymorphism affected H. pylori eradication rate and to investigate the effect of H. pylori eradication on angiogenesis and the inflammatory process according to the IL-8-251 polymorphism. A total of 250 H. pylori-positive patients treated by endoscopic resection of the gastric neoplasm were classified into 3 groups (134 H. pylori-eradicated group, 19 H. pylori-eradication failure group, and 97 H. pylori-infected group). H. pylori status, histology, and angiogenic factor levels were evaluated at baseline, 6 months, and 18 months. H. pylori eradication rate was 92.9% in AA genotype, 85.7% in AT genotype and 88.4% in TT genotype (P value = 0.731). Elevated IL-8 and matrix metalloproteinase-9 concentrations in H. pylori-infected gastric mucosa were reversible by successful eradication of H. pylori, independent of the IL-8-251 polymorphism. It is suggested that elevated IL-8 and MMP-9 concentrations in H. pylori-infected gastric mucosa are altered significantly after successful eradication and these conditions continue for 18 months. However, IL-8-251 polymorphism does not affect H. pylori eradication rate and the sequential changes of related angiogenic factors after H. pylori eradication in Koreans.

Topics: Aged; Alleles; Angiopoietin-1; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Asian People; Female; Gastric Mucosa; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Matrix Metalloproteinase 9; Middle Aged; Polymorphism, Single Nucleotide; Proton Pump Inhibitors; Republic of Korea; Retrospective Studies; Stomach Neoplasms; Time Factors; Vascular Endothelial Growth Factor A

Interleukin-8 (IL-8) plays a central role in the pathogenesis of Helicobacter pylori infection. We used four different H. pylori strains isolated from patients with gastritis or duodenal ulcer disease to examine their differential effects on signaling pathways and IL-8 gene response in gastric epithelial cells. IL-8 mRNA level is elevated in response to high (100) multiplicity of infection (MOI) independent of cagA, vacA, and dupA gene characteristics. By lower MOIs (1 or 10), only cagA ( + ) strains significantly induce IL-8 gene expression. This is based on differential regulation of IL-8 promoter activity. Analysis of intracellular signaling pathways indicates that H. pylori clinical isolates induce IL-8 gene transcription through NF-κB p65, but by a MOI-dependent differential activation of MAPK pathways. Thus, the major virulence factors of H. pylori CagA, VacA, and DupA might play a minor role in the level of IL-8 gene response to a high bacterial load.

Topics: Bacterial Load; Bacterial Proteins; Cell Line; Duodenal Ulcer; Epithelial Cells; Gastritis; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mitogen-Activated Protein Kinase Kinases; NF-kappa B; Promoter Regions, Genetic; Signal Transduction; Stomach; Transcription, Genetic; Virulence Factors

The dupA of Helicobacter pylori has been suggested as a virulence marker associated with the development of duodenal ulcer disease. However, the studies performed in different geographical areas have shown that there are variations in the prevalence of dupA and its association with H. pylori clinical outcomes. Our group did not observe associations between the presence of dupA and H. pylori clinical outcomes in Brazil. On the other hand, we observed 2 mutations in the sequence of dupA that lead to stop codons: a deletion of an adenine at position 1311 and an insertion of an adenine at position 1426 of the gene. Our aim was to evaluate associations of the presence of dupA with duodenal ulcer and gastric cancer, considering dupA-positive only those H. pylori strains that do not have the mutations in the gene sequence. We also evaluated the effect of infection with a strain carrying an intact dupA on the gastric mucosa histology and IL-8 gastric levels. Colonization with strains that had the intact dupA was negatively associated with gastric carcinoma (p=0.001, OR=0.32, 95% CI=0.16-0.66). The presence of dupA was also associated with an increased degree of antral mucosa inflammation (p=0.01) and with decreased corpus atrophy (p<0.01) as well as with increased gastric mucosa IL-8 levels (p=0.04). In conclusion, the infection with a H. pylori strain containing the dupA without the stop codon polymorphisms is associated with a lower risk of development of gastric carcinoma in Brazilian subjects.

Topics: Adult; Aged; Brazil; Duodenal Ulcer; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Polymorphism, Genetic; Stomach Neoplasms; Virulence Factors

Helicobacter pylori is a Gram-negative, spiral-shaped microorganism associated with acute and chronic gastritis, peptic ulcer, gastric cancer and gastric lymphomas in humans. H. pylori neutrophil-activating protein (NAP) is a major virulence factor playing a central role in pathogenesis of mucosal inflammation by immune cell attraction and Th1 cytokine response polarization. NAP is protective antigen and promising vaccine candidate against H. pylori infection. Here we present the development of measles virus (MV) vaccine strain encoding the NAP antigen. In order to facilitate the extracellular transport and detection, NAP was inserted in the human lambda immunoglobulin chain replacing a major part of the variable domain. We generated two MV vectors expressing secretory NAP forms: MV-lambda-NAP encoding the full-length constant lambda light chain domain and MV-s-NAP encoding only the N-terminus of the lambda light chain with the leader peptide. Immunization of MV permissive Ifnarko-CD46Ge transgenic mice by a single intraperitoneal injection of the NAP-expressing strains induced a robust, long-term humoral and cellular immune response against MV. Nine months post vaccination measles-neutralizing antibody titers were above the serum level considered protective for humans. Furthermore, all animals immunized with MV strains expressing the secretory NAP antigen developed strong humoral immunity against NAP, reaching titers >1:10,000 within 2-4 weeks. IFN-γ ELISpot assay confirmed that NAP-encoding MV vectors can also stimulate NAP-specific cell-mediated immunity. Our data demonstrate that MV is an excellent vector platform for expression of bacterial antigens and development of vaccines for H. pylori immunoprophylaxis in humans.

Topics: Animals; Antibodies, Bacterial; Antibodies, Monoclonal; Antibodies, Neutralizing; Antibodies, Viral; Antigens, Bacterial; Bacterial Proteins; Bacterial Vaccines; Cell Line, Tumor; Chlorocebus aethiops; Female; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Humoral; Interferon-gamma; Interleukin-8; Measles; Measles virus; Mice; Mice, Knockout; Mice, Transgenic; Neutralization Tests; Recombinant Proteins; Vaccines, Attenuated; Vero Cells

Helicobacter pylori infects half the world's population, and carriage is lifelong without antibiotic therapy. Current regimens prescribed to prevent infection-associated diseases such as gastroduodenal ulcers and gastric cancer can be thwarted by antibiotic resistance. We reported that administration of 1% D,L-α-difluoromethylornithine (DFMO) to mice infected with H. pylori reduces gastritis and colonization, which we attributed to enhanced host immune response due to inhibition of macrophage ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis. Although no ODC has been identified in any H. pylori genome, we sought to determine if DFMO has direct effects on the bacterium. We found that DFMO significantly reduced the growth rate of H. pylori in a polyamine-independent manner. Two other gram-negative pathogens possessing ODC, Escherichia coli and Citrobacter rodentium, were resistant to the DFMO effect. The effect of DFMO on H. pylori required continuous exposure to the drug and was reversible when removed, with recovery of growth rate in vitro and the ability to colonize mice. H. pylori exposed to DFMO were significantly shorter in length than those untreated and they contained greater internal levels of ATP, suggesting severe effects on bacterial metabolism. DFMO inhibited expression of the H. pylori virulence factor cytotoxin associated gene A, and its translocation and phosphorylation in gastric epithelial cells, which was associated with a reduction in interleukin-8 expression. These findings suggest that DFMO has effects on H. pylori that may contribute to its effectiveness in reducing gastritis and colonization and may be a useful addition to anti-H. pylori therapies.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Cells, Cultured; Eflornithine; Enzyme Inhibitors; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Mice; Mice, Inbred C57BL; Protein Transport; Transcriptional Activation

Helicobacter pylori is an important class I carcinogen that persistently infects the human gastric mucosa to induce gastritis, gastric ulceration, and gastric cancer. H. pylori pathogenesis strongly depends on pathogenic factors, such as VacA (vacuolating cytotoxin A) or a specialized type IV secretion system (T4SS), which injects the oncoprotein CagA (cytotoxin-associated gene A product) into the host cell. Since access to primary gastric epithelial cells is limited, many studies on the complex cellular and molecular mechanisms of H. pylori were performed in immortalized epithelial cells originating from individual human adenocarcinomas. The aim of our study was a comparative analysis of 14 different human gastric epithelial cell lines after colonization with H. pylori. We found remarkable differences in host cell morphology, extent of CagA tyrosine phosphorylation, adhesion to host cells, vacuolization, and interleukin-8 (IL-8) secretion. These data might help in the selection of suitable cell lines to study host cell responses to H. pylori in vitro, and they imply that different host cell factors are involved in the determination of H. pylori pathogenesis. A better understanding of H. pylori-directed cellular responses can provide novel and more balanced insights into the molecular mechanisms of H. pylori-dependent pathogenesis in vivo and may lead to new therapeutic approaches.

Topics: Adenocarcinoma; Bacterial Translocation; Cell Adhesion; Cell Line, Tumor; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Immunoblotting; Immunoprecipitation; Interleukin-8; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms

Anti-Helicobacter pylori heat shock protein 60 (HpHSP60) antibodies are usually found in H. pylori-infected patients and are known to be associated with the progression of gastric diseases. However, the effects of these antibodies on the functions of HpHSP60 have not been identified. This study aims to investigate the effects of the interaction between anti-HSP60 antibodies and HpHSP60 on inflammatory responses. Anti-HpHSP60 polyclonal sera and monoclonal antibodies (mAbs) were produced to evaluate their effects on HpHSP60-induced IL-8 and TNF-α activity. The results indicated that anti-HpHSP60 polyclonal sera collected from patients infected with H. pylori or from rabbit and mice immunized with HpHSP60 could significantly enhance HpHSP60-mediated IL-8 and TNF-α secretion from monocytic THP-1 cells. Similar effects were also found with anti-HpHSP60 mAbs. Further analysis revealed that this phenomenon was only carried out by anti-HpHSP60 antibody but not by other non-specific mAbs. Moreover, the non-specific mAbs decreased the synergism of HpHSP60 and anti-HpHSP60 mAbs in proinflammatory cytokine induction. Herein, we have examined the role of anti-HpHSP60 antibody in host immune responses for the first time. This study demonstrated that H. pylori HSP60/mAbs could modulate helicobacterial pathogenesis by increasing IL-8 and TNF-α production. The pathogen-specific antibodies may execute potential immune functions rather than recognize or neutralize microbes.

Topics: Animals; Antibodies; Bacterial Proteins; Cell Line; Chaperonin 60; Female; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Mice; Mice, Inbred BALB C; Rabbits; Receptors, Fc; Signal Transduction; Tumor Necrosis Factor-alpha

Helicobacter pylori is the causative agent for developing gastritis, gastric ulcer, and even gastric cancer. Virulent strains carry the cag pathogenicity island (cagPAI) encoding a type-IV secretion system (T4SS) for injecting the CagA protein. However, mechanisms of sensing this pathogen through Toll-like receptors (TLRs) and downstream signalling pathways in the development of different pathologies are widely unclear. Here, we explored the involvement of TLR-2 and TLR-5 in THP-1 cells and HEK293 cell lines (stably transfected with TLR-2 or TLR-5) during infection with wild-type H. pylori and isogenic cagPAI mutants. H. pylori triggered enhanced TLR-2 and TLR-5 expression in THP-1, HEK293-TLR2 and HEK293-TLR5 cells, but not in the HEK293 control. In addition, IL-8 and TNF-α cytokine secretion in THP-1 cells was induced in a cagPAI-dependent manner. Furthermore, we show that HEK293 cells are not competent for the uptake of T4SS-delivered CagA, and are therefore ideally suited for studying TLR signalling in the absence of T4SS functions. HEK293 control cells, which do not induce TLR-2 and TLR-5 expression during infection, only secreted cytokines in small amounts, in agreement with T4SS functions being absent. In contrast, HEK293-TLR2 and HEK293-TLR5 cells were highly competent for inducing the secretion of IL-8 and TNF-α cytokines in a cagPAI-independent manner, suggesting that the expression of TLR-2 or TLR-5 has profoundly changed the capability to trigger pro-inflammatory signalling upon infection. Using phospho-specific antibodies and luciferase reporter assays, we further demonstrate that H. pylori induces IRAK-1 and IκB phosphorylation in a TLR-dependent manner, and this was required for activation of transcription factor NF-κB. Finally, NF-κB activation in HEK293-TLR2 and HEK293-TLR5 cells was confirmed by expressing p65-GFP which was translocated from the cytoplasm into the nucleus. These data indicate that H. pylori-induced expression of TLR-2 and TLR-5 can qualitatively shift cagPAI-dependent to cagPAI-independent pro-inflammatory signalling pathways with possible impact on the outcome of H. pylori-associated diseases.

Topics: Antigens, Bacterial; Bacterial Proteins; Cell Line; Genomic Islands; HEK293 Cells; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Proteins; Inflammation Mediators; Interleukin-1 Receptor-Associated Kinases; Interleukin-8; NF-kappa B; Phosphorylation; Phosphoserine; RNA, Messenger; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 5; Transcription Factor AP-1; Transcriptional Activation; Tumor Necrosis Factor-alpha; Up-Regulation

Chronic infection of Helicobacter pylori in the stomach mucosa with translocation of the bacterial cytotoxin-associated gene A (CagA) effector protein via the cag-Type IV secretion system (TFSS) into host epithelial cells are major risk factors for gastritis, gastric ulcers, and cancer. The blood group antigen-binding adhesin BabA mediates the adherence of H. pylori to ABO/Lewis b (Le(b)) blood group antigens in the gastric pit region of the human stomach mucosa. Here, we show both in vitro and in vivo that BabA-mediated binding of H. pylori to Le(b) on the epithelial surface augments TFSS-dependent H. pylori pathogenicity by triggering the production of proinflammatory cytokines and precancer-related factors. We successfully generated Le(b)-positive cell lineages by transfecting Le(b)-negative cells with several glycosyltransferase genes. Using these established cell lines, we found increased mRNA levels of proinflammatory cytokines (CCL5 and IL-8) as well as precancer-related factors (CDX2 and MUC2) after the infection of Le(b)-positive cells with WT H. pylori but not with babA or TFSS deletion mutants. This increased mRNA expression was abrogated when Le(b)-negative cells were infected with WT H. pylori. Thus, H. pylori can exploit BabA-Le(b) binding to trigger TFSS-dependent host cell signaling to induce the transcription of genes that enhance inflammation, development of intestinal metaplasia, and associated precancerous transformations.

Topics: Adhesins, Bacterial; Animals; Bacterial Adhesion; Bacterial Secretion Systems; Chemokine CCL5; CHO Cells; Cricetinae; Cricetulus; Dogs; Gastric Mucosa; Gene Deletion; Helicobacter Infections; Helicobacter pylori; Homeodomain Proteins; Humans; Inflammation; Interleukin-8; Lewis Blood Group Antigens; Metaplasia; Mucin-2; Precancerous Conditions; Signal Transduction

Although control of the host cytokine network is known to influence gastric cancer susceptibility, the specific inflammatory responses in gastric carcinogenesis remain unclear. We prospectively examined the relationships between gastric cancer risk and plasma levels of interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor (TNF)-α in a nested case control study within The Shanghai Women's Health Study. Two controls were matched to each case on the basis of age, menopausal status, and sample collection parameters. The associations between gastric cancer risk and tertiles of cytokine levels were estimated by odds ratios (OR) and 95% confidence intervals (CI) from conditional logistic regression, adjusting for education. During a median follow-up period of 4 years (range 0.1-8 years), 141 women developed gastric cancer and were matched to 282 cancer-free study participants. Elevated levels of plasma IL-6 were associated with an increased risk of gastric cancer (P(trend) = 0.04). Risk increased 70% (OR = 1.7; 95% CI 1.0, 3.0) for women in the highest tertile (>4 pg/mL) of IL-6 compared with those in the lowest tertile (<1.8 pg/mL). The association between gastric cancer risk and IL-6 was stronger after 4 years of follow-up (OR = 2.6 [95% CI 1.0, 6.7] for highest versus lowest tertile) compared with an OR of 1.4 (95% CI 0.7, 2.9) for those diagnosed within 1-4 years of follow-up. No associations were observed with the other pro-inflammatory cytokines examined, namely IL-1β, IL-8, and TNF-α. Systemic plasma IL-6 levels may inform long-term gastric cancer risk. This novel finding awaits confirmation in future studies with sequential plasma collection.

Topics: Adult; Aged; Case-Control Studies; China; Confidence Intervals; Cytokines; Eating; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Middle Aged; Odds Ratio; Prospective Studies; Risk Factors; Stomach Neoplasms; Surveys and Questionnaires; Tumor Necrosis Factor-alpha

Paxillin is involved in the regulation of Helicobacter pylori-mediated gastric epithelial cell motility. We investigated the signaling pathways regulating H. pylori-induced paxillin phosphorylation and the effect of the H. pylori virulence factors cag pathogenicity island (PAI) and outer inflammatory protein (OipA) on actin stress fiber formation, cell phenotype, and IL-8 production. Gastric cell infection with live H. pylori induced site-specific phosphorylation of paxillin tyrosine (Y) 31 and Y118 in a time- and concentration-dependent manner. Activated paxillin localized in the cytoplasm at the tips of H. pylori-induced actin stress fibers. Isogenic oipA mutants significantly reduced paxillin phosphorylation at Y31 and Y118 and reduced actin stress fiber formation. In contrast, cag PAI mutants only inhibited paxillin Y118 phosphorylation. Silencing of epidermal growth factor receptor (EGFR), focal adhesion kinase (FAK), or protein kinase B (Akt) expression by small-interfering RNAs or inhibiting kinase activity of EGFR, Src, or phosphatidylinositol 3-kinase (PI3K) markedly reduced H. pylori-induced paxillin phosphorylation and morphologic alterations. Reduced FAK expression or lack of Src kinase activity suppressed H. pylori-induced IL-8 production. Compared with infection with the wild type, infection with the cag PAI mutant and oipA mutant reduced IL-8 production by nearly 80 and 50%. OipA-induced IL-8 production was FAK- and Src-dependent, although a FAK/Src-independent pathway for IL-8 production also exists, and the cag PAI may be mainly involved in this pathway. We propose paxillin as a novel cellular target for converging H. pylori-induced EGFR, FAK/Src, and PI3K/Akt signaling to regulate cytoskeletal reorganization and IL-8 production in part, thus contributing to the H. pylori-induced diseases.

Topics: Actins; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Cell Line; Epithelial Cells; ErbB Receptors; Focal Adhesion Kinase 1; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Paxillin; Phosphatidylinositol 3-Kinases; Phosphorylation; Signal Transduction

Population genetic analyses of bacterial genes whose products interact with host tissues can give new understanding of infection and disease processes. Here we show that strains of the genetically diverse gastric pathogen Helicobacter pylori from Amerindians from the remote Peruvian Amazon contain novel alleles of cagA, a major virulence gene, and reveal distinctive properties of their encoded CagA proteins. CagA is injected into the gastric epithelium where it hijacks pleiotropic signaling pathways, helps Hp exploit its special gastric mucosal niche, and affects the risk that infection will result in overt gastroduodenal diseases including gastric cancer. The Amerindian CagA proteins contain unusual but functional tyrosine phosphorylation motifs and attenuated CRPIA motifs, which affect gastric epithelial proliferation, inflammation, and bacterial pathogenesis. Amerindian CagA proteins induced less production of IL-8 and cancer-associated Mucin 2 than did those of prototype Western or East Asian strains and behaved as dominant negative inhibitors of action of prototype CagA during mixed infection of Mongolian gerbils. We suggest that Amerindian cagA is of relatively low virulence, that this may have been selected in ancestral strains during infection of the people who migrated from Asia into the Americas many thousands of years ago, and that such attenuated CagA proteins could be useful therapeutically.

Topics: Alleles; Amino Acid Motifs; Amino Acid Sequence; Animals; Antigens, Bacterial; Bacterial Proteins; Evolution, Molecular; Female; Gastric Mucosa; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Humans; Indians, South American; Interleukin-8; Male; Molecular Sequence Data; Mucin-2; Peru; Phosphorylation; Signal Transduction; Stomach Neoplasms; Virulence Factors

Helicobacter pylori induces cytokine mediated changes in gastroduodenal pathophysiology, wherein, the activated macrophages at the sub-mucosal space play a central role in mounting innate immune response against the antigens. The bacterium gains niche through persistent inflammation and local immune-suppression causing peptic ulcer disease or chronic gastritis; the latter being a significant risk factor for the development of gastric adenocarcinoma. What favors persistence of H. pylori in the gastric niches is not clearly understood. We report detailed characterization of a functionally unknown gene (HP986), which was detected in patient isolates associated with peptic ulcer and gastric carcinoma. Expression and purification of recombinant HP986 (rHP986) revealed a novel, ∼29 kDa protein in biologically active form which associates with significant levels of humoral immune responses in diseased individuals (p<0.001). Also, it induced significant levels of TNF-α and Interleukin-8 in cultured human macrophages concurrent to the translocation of nuclear transcription factor-κB (NF-κB). Further, the rHP986 induced apoptosis of cultured macrophages through a Fas mediated pathway. Dissection of the underlying signaling mechanism revealed that rHP986 induces both TNFR1 and Fas expression to lead to apoptosis. We further demonstrated interaction of HP986 with TNFR1 through computational and experimental approaches. Independent proinflammatory and apoptotic responses triggered by rHP986 as shown in this study point to its role, possibly as a survival strategy to gain niche through inflammation and to counter the activated macrophages to avoid clearance.

Topics: Amino Acid Sequence; Apoptosis; Bacterial Proteins; fas Receptor; Gastrointestinal Diseases; Genetic Loci; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Humoral; Inflammation Mediators; Interleukin-8; Models, Biological; Molecular Sequence Data; NF-kappa B; Protein Binding; Receptors, Tumor Necrosis Factor, Type I; Recombinant Proteins; Sequence Alignment; Sequence Analysis, Protein; Time Factors; Tumor Necrosis Factor-alpha

Literature data indicate an association between the presence of Helicobacter spp. in the liver and the development of hepatocellular carcinoma (HCC). However, the role of H. pylori infections in chronic liver diseases (CLD) remains controversial. The aim of this study was to detect Helicobacter spp. DNA in patients with CLD, and to investigate the host response to the presence of the bacterium in the liver. Helicobacter spp. DNA was detected in 59% samples. H.pylori was the most prevalent species (94%). We estimated the expression level of IL-1 and IL-8 genes. The presence of Helicobacter spp. did not have a significant effect on the gene expression of IL-8 and IL-1.

Topics: Biopsy; Carcinoma, Hepatocellular; Chronic Disease; DNA, Bacterial; DNA, Ribosomal; Helicobacter; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-1; Interleukin-8; Liver Diseases; Liver Neoplasms; Poland; Polymerase Chain Reaction; RNA; RNA, Ribosomal, 16S; Sequence Analysis, DNA

Protease-activated receptors (PAR) are seven transmembrane receptors that are expressed throughout the gastrointestinal tract. In vitro experiments using gastric tumor cell lines, murine models and one clinical study provided evidence for a potential role of PAR2 in Helicobacter pylori-induced gastritis.. To investigate PAR2 expression in H. pylori-infected patients and correlation with proinflammatory IL-8, IL-1β as well as histologic changes of the mucosa. Furthermore, PAR2 expression was studied in context to mucosal amounts of secretory leukocyte protease inhibitor (SLPI), a putative regulator of PAR2.. Twenty-two H. pylori-infected patients and 72 H. pylori-negative subjects underwent upper GI endoscopy. In antrum-derived mucosal biopsies, PAR2, IL-1β, IL-8, and SLPI expression was analyzed by quantitative RT-PCR, and in part by ELISA and immunohistochemistry. Histopathologic evaluation of gastritis was performed according to the updated Sydney classification.. IL-8 gene expression was 5-fold increased in the mucosa of H. pylori-infected patients compared with non-infected (p < .0001), whereas no differences for PAR2 and IL-1β mRNA amounts were observed between both groups. PAR2 gene expression correlated positively with transcript levels of IL-8, IL-1β as well mucosal SLPI levels in H. pylori-infected patients (r: 0.47-0.84; p < .0001), whereas no correlation was found with the degree of gastritis.. PAR2 represents an additive pathway of IL-8 secretion and proinflammatory effects in H. pylori-induced gastritis. Reduced SLPI levels leading to higher serine protease activities in the mucosa of infected subjects might regulate PAR2 activation.

Topics: Adult; Animals; Female; Gastric Mucosa; Gastritis; Gene Expression Profiling; Helicobacter Infections; Helicobacter pylori; Histocytochemistry; Host-Pathogen Interactions; Humans; Immunohistochemistry; Interleukin-1beta; Interleukin-8; Male; Mice; Middle Aged; Receptor, PAR-2

There are few data on circulatory pro-inflammatory cytokine levels and cytokine gene polymorphisms in H. pylori-positive patients. A cross-sectional study was conducted to examine the effects of H. pylori infection, gastric atrophy, and the IL-8 T-251A polymorphism on plasma IL-8 levels in 98 Japanese adults. Seventy-one subjects were positive for H. pylori infection. The geometric mean of plasma IL-8 concentration was significantly higher in subjects with H. pylori infection than in those without (P=0.001). The development of atrophy was negatively associated with IL-8 levels in the H. pylori-positive subjects, although not significantly. Plasma IL-8 levels in the T/T genotype were associated with H. pylori infection and atrophy status (P=0.016). Our findings suggested that circulating IL-8 levels were associated with H. pylori infection. The effect of H. pylori infection on plasma IL-8 levels was not clearly modified by the IL-8 T-251A polymorphism.

Topics: Adult; Aged; Atrophy; Cross-Sectional Studies; Female; Gastric Mucosa; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Japan; Male; Middle Aged; Polymorphism, Genetic; Young Adult

The dupA gene of Helicobacter pylori was suggested to be a risk factor for duodenal ulcer but protective against gastric cancer. The present study aimed to re-examine the role of dupA in H. pylori-infected Japanese patients. We found that dupA status was not associated with any gastroduodenal disease, histological score of chronic gastritis or with the extent of interleukin-8 production from gastric cell lines. These results indicate that dupA is unlikely to be a virulence factor of H. pylori in the Japanese population.

Topics: Adult; Aged; Aged, 80 and over; Asian People; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Severity of Illness Index; Stomach; Treatment Outcome; Virulence Factors

Helicobacter pylori-related disease is at least partially attributable to the genotype of the infecting strain, particularly the presence of specific virulence factors. We investigated the prevalence of a novel combination of H. pylori virulence factors, including the cag pathogenicity island (PAI), and their association with severe disease in isolates from the three major ethnicities in Malaysia and Singapore, and evaluated whether the cag PAI was intact and functional in vitro. Polymerase chain reaction (PCR) was used to detect dupA, cagA, cagE, cagT, cagL and babA, and to type vacA, the EPIYA motifs, HP0521 alleles and oipA ON status in 159 H. pylori clinical isolates. Twenty-two strains were investigated for IL-8 induction and CagA translocation in vitro. The prevalence of cagA, cagE, cagL, cagT, babA, oipA ON and vacA s1 and i1 was >85%, irrespective of the disease state or ethnicity. The prevalence of dupA and the predominant HP0521 allele and EPIYA motif varied significantly with ethnicity (p < 0.05). A high prevalence of an intact cag PAI was found in all ethnic groups; however, no association was observed between any virulence factor and disease state. The novel association between the HP0521 alleles, EPIYA motifs and host ethnicity indicates that further studies to determine the function of this gene are important.

Topics: Adult; Antigens, Bacterial; Bacterial Proteins; Cells, Cultured; DNA, Bacterial; Epithelial Cells; Genetic Variation; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Malaysia; Polymerase Chain Reaction; Protein Transport; Singapore; Virulence Factors

Polymorphisms in cytokine genes may contribute to increased susceptibility to different cancers. The aim of this paper is to investigate the association of IL-8-251A/T polymorphism and Helicobacter pylori (H. pylori) infection with the risk of developing gastric cardiac adenocarcinoma (GCA) in the south of Taihang Mountain, a high-incidence area of esophageal cancer in China. The IL-8-251 A/T polymorphism was genotyped in 519 cases of GCA and 504 healthy controls. The H. pylori infection in GCA patients and controls was detected by rapid urease test (RUT), histopathology or (14)C-urea breath test ((14)C-UBT). The results showed that family history of upper gastrointestinal cancer (UGIC) and H. pylori infection significantly increased the risk of developing GCA. The overall genotype and allelotype distributions of IL-8 promoter SNPs in GCA patients were significantly different from those in healthy controls. Compared with TT genotype, AA genotype significantly elevated the risk of developing GCA. The stratification analysis revealed that, compared with the TT genotype, the AA genotype significantly elevated the risk of developing GCA in both positive family history of UGIC and H. pylori infection subgroups. This study provides evidence to support a relationship of increased susceptibility to GCA in individuals of the south Taihang Mountain region with IL-8 251 AA genotype, especially for those individuals who have family history of UGIC or H. pylori infection.

Topics: Adenocarcinoma; Aged; Base Sequence; China; Demography; Female; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Incidence; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Precancerous Conditions; Risk Factors; Sequence Analysis, DNA; Stomach Neoplasms

gamma-Glutamyl transpeptidase (GGT) has been reported to be a virulence factor of Helicobacter pylori associated with bacterial colonization and cell apoptosis. But its mechanism of pathogenesis is not firmly established. This study aims to examine its role in H pylori-mediated infection.. Various H pylori isogenic mutants were constructed by a polymerase chain reaction (PCR) approach. H pylori native GGT protein (HP-nGGT) was purified with ion-exchange and gel-filtration chromatography. Generation of H2O2 was measured with fluorimetric analysis, whereas nuclear factor-kappaB (NF-kappaB) activation was determined by luciferase assay and Western blot. Cytokine production was examined by enzyme-linked immunoabsorbent assay and real-time PCR. DNA damage was assessed with comet assay and flow cytometry. The GGT activity of 98 H pylori isolates was analyzed by an enzymatic assay.. Purified HP-nGGT generated H2O2 in primary gastric epithelial cells and AGS gastric cancer cells, resulting in the activation of NF-kappaB and up-regulation of interleukin-8 (IL-8) production. In addition, HP-nGGT caused an increase in the level of 8-OH-dG, indicative of oxidative DNA damage. In contrast, Deltaggt showed significantly reduced levels of H2O2 generation, IL-8 production, and DNA damage in cells compared with the wild type (P<.05). The clinical importance of GGT was indicated by significantly higher (P<.001) activity in H pylori isolates obtained from patients with peptic ulcer disease (n=54) than isolates from patients with nonulcer dyspepsia (n=44).. Our findings provide evidence that GGT is a pathogenic factor associated with H pylori-induced peptic ulcer disease.

Topics: Apoptosis; Bacterial Proteins; Biopsy; Blotting, Western; Cells, Cultured; Chromatography, Gel; Chromatography, Ion Exchange; Comet Assay; DNA Damage; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fluorometry; gamma-Glutamyltransferase; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen Peroxide; Interleukin-8; Mutation; NF-kappa B; Oxidative Stress; Reverse Transcriptase Polymerase Chain Reaction; Stomach Ulcer; Time Factors; Virulence Factors

The purpose of this paper is to investigate the relationship between clinical outcome and the intactness of cagPAI in Helicobacter pylori strains from Vietnam. The presence or absence of 30 cagPAI genes was investigated by polymerase chain reaction (PCR) and dot-blotting. H. pylori-induced interleukin-8 secretion and hummingbird phenotype, and H. pylori adhesion to gastric epithelial cells were examined. The serum concentration of pepsinogen 1, pepsinogen 2, and gastrin was also measured in all patients. cagPAI was present in all 103 Vietnamese H. pylori isolates, of which 91 had intact cagPAI and 12 contained only a part of cagPAI. Infection with the partial cagPAI strains was less likely to be associated with peptic ulcer and chronic gastric mucosal inflammation than infection with strains possessing intact cagPAI. The partial cagPAI strains lacked almost all ability to induce interleukin-8 secretion and the hummingbird phenotype in gastric cells. Their adhesion to epithelial cells was significantly decreased in comparison with intact cagPAI strains. Moreover, for the first time, we found an association between cagPAI status and the serum concentration of pepsinogens 1 and 2 in infected patients. H. pylori strains with internal deletion within cagPAI are less virulent and, thus, less likely to be associated with severe clinical outcomes.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bacterial Adhesion; Bacterial Proteins; DNA, Bacterial; Epithelial Cells; Female; Gastrins; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Pepsinogen A; Peptic Ulcer; Polymerase Chain Reaction; Polymorphism, Genetic; Vietnam; Virulence; Virulence Factors; Young Adult

To investigate the effects of interleukin-8 (IL-8), macrophage migration inhibitory factor (MIF) gene polymorphisms, Helicobacter pylori (H. pylori) infection, on the risk of developing severe chronic atrophic gastritis (SCAG) and intestinal metaplasia (IM).. A total of 372 cases were selected from a cohort study in Linqu County, a high risk area for gastric cancer (GC) in northern China. To obtain a sufficient group size, patients with normal or superficial gastritis were included. Based on an average follow-up period of 56 mo, the 372 cases were divided into no progression group (no histological progression from normal or superficial gastritis, n = 137), group I (progressed from normal or superficial gastritis to SCAG, n = 134) and group II (progressed from normal or superficial gastritis to IM, n = 101). IL-8, MIF gene polymorphisms were detected by polymerase chain reaction-based denaturing high-performance liquid chromatography analysis and DNA sequencing.. An increased risk of SCAG was found in subjects with IL-8-251 AA genotype [odds ratio (OR) = 2.62, 95% CI: 1.23-5.72] or IL-8-251 A allele carriers (AA + AT) (OR = 1.81, 95% CI: 1.06-3.09). An elevated risk of IM was found in subjects with IL-8-251 AT genotype (OR = 2.27, 95% CI: 1.25-4.14) or IL-8-251 A allele carriers (OR = 2.07, 95% CI: 1.16-3.69). An increased risk of SCAG was found in subjects with MIF-173 GC genotype (OR = 2.36, 95% CI: 1.38-4.02) or MIF-173 C allele carriers (GC + CC) (OR = 2.07, 95% CI: 1.21-3.55). An elevated risk of IM was found in subjects with MIF-173 CC genotype (OR = 2.27, 95% CI: 1.16-4.46) or MIF-173 C allele carriers (OR = 3.84, 95% CI: 1.58-9.34). The risk of SCAG and IM was more evident in subjects carrying IL-8-251 A allele (OR = 6.70, 95% CI: 1.29-9.78) or MIF-173 C allele (OR = 6.54, 95% CI: 2.97-14.20) and positive for H. pylori infection.. IL-8-251 and MIF-173 gene polymorphisms are significantly associated with the risk of SCAG and IM in a population with a high risk of GC in Linqu County, Shandong Province, China.

Topics: Adult; Aged; Cohort Studies; Cytokines; Female; Gastritis, Atrophic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestines; Intramolecular Oxidoreductases; Macrophage Migration-Inhibitory Factors; Male; Metaplasia; Middle Aged; Polymorphism, Genetic; Risk Factors; Stomach Neoplasms; Young Adult

Helicobacter pylori infection of the gastric mucosa causes an active-chronic inflammation that is strongly linked to the development of duodenal and gastric ulcers and stomach cancer. However, greater than 80% of individuals infected with H. pylori are asymptomatic beyond histologic inflammation, and it is unknown what factors influence the incidence and character of bacterial-associated gastritis and related disorders. Because previous studies demonstrated that the Muc1 epithelial glycoprotein inhibited inflammation during acute lung infection by Pseudomonas aeruginosa, we asked whether Muc1 might also counter-regulate gastric inflammation in response to H. pylori infection. Muc1(-/-) mice displayed increased bacterial colonization of the stomach and greater TNF-alpha and keratinocyte chemoattractant transcript levels compared with Muc1(+/+) mice after experimental H. pylori infection. Knockdown of Muc1 expression in AGS human gastric epithelial cells by RNA interference was associated with increased phosphorylation of IkappaBalpha, augmented activation and nuclear translocation of NF-kappaB, and enhanced production of interleulin-8 compared with Muc1-expressing cells. Conversely, Muc1 overexpression was correlated with decreased NF-kappaB activation, reduced interleulin-8 production, and diminished IkappaB kinase beta (IKKbeta)/IKKgamma coimmunoprecipitation compared with cells expressing Muc1 endogenously. Cotransfection of AGS cells with Muc1 plus IKKbeta, but not a catalytically inactive IKKbeta mutant, reversed the Muc1 inhibitory effect. Finally, Muc1 formed a coimmunoprecipitation complex with IKKgamma but not with IKKbeta. These results are consistent with the hypothesis that Muc1 binds to IKKgamma, thereby inhibiting formation of the catalytically active IKK complex and blocking the ability of H. pylori to stimulate IkappaBalpha phosphorylation, NF-kappaB activation, and downstream inflammatory responses.

Topics: Animals; Cholera Toxin; Colony-Forming Units Assay; DNA; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Gene Amplification; Gene Expression Regulation; Gene Knockdown Techniques; Genes, Reporter; Helicobacter Infections; Helicobacter pylori; I-kappa B Kinase; I-kappa B Proteins; Interleukin-8; Male; Mice; Mice, Knockout; Mucin-1; NF-KappaB Inhibitor alpha; Recombination, Genetic; Reverse Transcriptase Polymerase Chain Reaction; Stomach; Transfection; Tumor Necrosis Factor-alpha

Infection with Helicobacter pylori possessing a newly described virulence factor--duodenal ulcer-promoting gene A (dupA)--has been associated with duodenal ulceration and increased gastric inflammation.. The dupA locus of 34 strains was sequenced. A panel of dupA mutants was generated and cocultured with human gastric epithelial cells and peripheral blood mononuclear cells; proinflammatory cytokine release was measured. IL8 expression was measured in human gastric biopsy specimens and related to the dupA and cagA status of infecting strains.. Most H. pylori strains had a dupA allele that was longer (1884 bp; dupA1) than previously described dupA alleles, although some had truncated versions (dupA2). Unlike the best-characterized H. pylori virulence determinant, the cag pathogenicity island (cag PaI), neither dupA type induced release of interleukin (IL)-8 from gastric epithelial cells. However, infections due to dupA-positive strains were associated with higher-level mucosal IL-8 messenger RNA expression in the human stomach than were infections due to dupA-negative strains. To explain this paradox, we found that dupA1 (but not dupA2 or the cag PaI) substantially increased H. pylori-induced IL-12p40 and IL-12p70 production from CD14(+) mononuclear cells. Other T helper 1-associated cytokines were also modestly induced.. We suggest that virulent H. pylori strains cause inflammation by stimulating epithelial cells through cag-encoded proteins and mononuclear inflammatory cells through dupA1 products.

Topics: Amino Acid Sequence; Base Sequence; Cell Line; Coculture Techniques; Cytokines; DNA Primers; Flow Cytometry; Gastric Mucosa; Gene Amplification; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Leukocytes, Mononuclear; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Polymorphism, Genetic; Sequence Alignment; Sequence Homology, Amino Acid; Virulence Factors

Previous studies imply that IL-1 and IL-8 gene variations may play a crucial role in the genetic predisposition to different gastric disorders upon H. pylori infection.. The aim of this study was to determine the potential association between the prevalence of certain polymorphic sites and the risk of gastric disorders in Iranian population.. One hundred and forty three unrelated individuals with different gastric disorders and 374 normal individuals with no gastric disorders and with a negative serology test for H. pylori (control group) were studied for the association between IL-1ß (+3953 C/T) and IL-8 (-251 A/T) gene polymorphisms and H. pylori-mediated gastritis and gastric ulcer. An analysis of genotype frequency for these genes was performed using RFLP-PCR.. Based on the data obtained from culture and pathologic findings, the patients were classified into three subpopulations: H. pylori(+) non-ulcerative gastritis(+), H. pylori(+) ulcerative gastritis(+) and H. pylori(-) non-ulcerative gastritis(+). A significantly higher frequency of TT genotype (p=0.02) in IL-1ß +3953 in H. pylori(+) ulcerative gastritis(+) was revealed compared to the control group. There were no significant differences among other subpopulations. No significant differences in allele and genotype frequencies of IL-8 (-251A/T) were found among the patients.. The data suggest that TT genotype in IL-1ß +3953 may be a major contributing genetic risk factor for H. pylori induced gastric ulcer. Moreover, the role of other bacterial and host response factors, such as bacterial adherence peptides, host chemokines, and genes involved in gastric acid secretion, must be further investigated in different ethnic populations.

Topics: DNA Mutational Analysis; Gastritis; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Haplotypes; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Polymorphism, Genetic; Stomach Ulcer

Helicobacter pylori (H. pylori) is an important risk factor of gastric adenocarcinoma. Interleukin (IL)-8 is a potent angiogenic factor and plays an important role in inflammation of gastric mucosa by H. pylori. Host susceptibility may help to predict H. pylori-infected individuals with a higher risk of gastric adenocarcinoma. The aim of this study was to clarify the effect of IL-8 polymorphism on angiogenesis in the process of gastric carcinogenesis in H. pylori-infected Koreans. The IL-8-251A/T polymorphism was genotyped by PCR-RFLP from a total of 395 subjects; 92 normal controls, 87 H. pylori-infected controls, 108 chronic atrophic gastritis and/or intestinal metaplasia and 108 adenocarcinoma. The gastric mucosal concentrations of IL-8, membrane metalloproteinase (MMP)-9, angiopoietin (Ang)-1, and vascular endothelial growth factor (VEGF) were measured by ELISA. MMP-9 concentrations were increased with disease progression. There was significant correlation between MMP-9 and disease progression in AA (r=0.42, p<0.01) and AT genotype (r=0.43, p<0.01). Ang-1 concentrations were increased in AA genotype (r=0.40, p=0.01). However, there was no significant correlation between VEGF and disease progression in AA genotype. In conclusion, IL-8-251 AA genotype may be associated with angiogenesis in gastric carcinogenesis in H. pylori-infected Koreans.

Topics: Adenocarcinoma; Adult; Aged; Angiopoietin-1; Asian People; Disease Progression; Female; Gastric Mucosa; Gastritis, Atrophic; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Korea; Male; Matrix Metalloproteinase 9; Metaplasia; Middle Aged; Neovascularization, Pathologic; Polymorphism, Genetic; Stomach Neoplasms; Vascular Endothelial Growth Factor A

Helicobacter pylori (H. pylori) infection induces cytokine production and is associated with gastrointestinal diseases. This study examined the relationship of gene polymorphisms, including interleukin (IL)-1beta, -10, -8, and tumor necrosis factor-alpha (TNF-alpha), H. pylori infection, and susceptibility to gastrointestinal disorders in Taiwanese patients.. IL-1beta-511/-31/+3953, -10-1082/-819/-592, -8-251, and TNF-alpha-308 polymorphisms were assessed in 628 gastrointestinal disease patients, and 176 healthy controls were analyzed using the polymerase chain reaction-restriction fragment length polymorphism method.. IL-1beta-511 T/T and -31 C/C genotypes, and IL-1beta-511 T and -31 C alleles were associated with an increased risk of reflux esophagitis (P = 0.034, odds ratio [OR] = 1.384, 95% confidence interval [CI]: 1.023-1.871; P = 0.031, OR = 1.388, 95% CI: 1.028-1.873; P = 0.044, OR = 1.342, 95% CI: 1.008-1.786; and P = 0.040, OR = 1.349, 95% CI: 1.014-1.796, respectively). No relationship was found between H. pylori infection and the risk of reflux esophagitis. IL-10-819 C/T and -10-592 A/C genotypes and IL-10-1082/-819/-592 ATA/ACC and ATA/GCC haplotypes were associated with an increased risk of gastritis (P = 0.021, OR = 1.721, 95% CI: 1.084-2.733; P = 0.016, OR = 1.766, 95% CI: 1.112-2.805; P = 0.039, OR = 1.662, 95% CI: 1.024-2.697; and P = 0.035, OR = 1.600, 95% CI: 1.024-2.499, respectively).. Among Taiwanese patients, IL-1beta and -10 polymorphisms were associated with an increased risk of erosive reflux esophagitis and gastritis, respectively.

Topics: Adult; Asian People; Case-Control Studies; Chi-Square Distribution; Esophagitis, Peptic; Female; Gastritis; Gene Frequency; Genetic Predisposition to Disease; Haplotypes; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-1beta; Interleukin-8; Linkage Disequilibrium; Logistic Models; Male; Middle Aged; Odds Ratio; Polymorphism, Genetic; Risk Assessment; Risk Factors; Taiwan; Tumor Necrosis Factor-alpha

Efforts to identify potent small molecule inhibitors of Helicobacter pylori led to the evaluation of 23 3',4',5'-trimethoxychalcone analogues. Some of the compounds displayed potent antibacterial activity against H. pylori. Three most active and selective compounds 1, 7, and 13 also showed the bactericide activity against the reference as well as multidrug-resistant strains of H. pylori. Additionally, the aforementioned three compounds potentially inhibited the H. pylori adhesion and invasion to human gastric epithelial (AGS) cells. Furthermore, these selective compounds inhibited the H. pylori-induced gastric inflammation by reduced inflammatory mediator's nuclear factor kappa B activation, and the secretion of interleukin-8.

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents; Cell Line; Chalcones; Epithelial Cells; Gastritis; Helicobacter Infections; Helicobacter pylori; Host-Pathogen Interactions; Humans; Interleukin-8; NF-kappa B

IceA of Helicobacter pylori (H. pylori) has been suggested as a virulence factor for the bacteria, but its pathogenic role remains unelucidated. Here, we examined the effect of iceA mutation on the secretion of IL-8 by human gastric epithelial cells. We also investigated whether the changes in IL-8 production caused by iceA mutation were associated with impaired adherence of H. pylori to the epithelial cells or with impaired apoptosis of these cells. The iceA mutant strain was constructed from wildtype H. pylori strain by insertional mutagenesis of iceA. The human gastric epithelial cells SGC7901 were infected with wildtype or mutant H. pylori for appropriate lengths of time. The adherence of the bacteria to the epithelial cells was examined by fluorescent microscopy using an anti-H. pylori antibody and flow cytometry. The apoptosis of the epithelial cells was studied by annexin-V staining and flow cytometry. The production of IL-8 by SGC 7901 cells was determined by ELISA. We found that iceA mutation was associated with significantly impaired production of IL-8 from the epithelial cells, which was not due to impaired adherence by the bacteria to the epithelial cells as wildtype and mutant H. pylori exhibited similar levels of binding to the epithelial cells. Furthermore, inactivation of iceA did not affect the apoptotic cell death of SGC7901. Our findings indicate that iceA may contribute to the pathogenicity of H. pylori by modulating the production of IL-8 by host epithelial cells.

Topics: Apoptosis; Bacterial Adhesion; Bacterial Outer Membrane Proteins; Cell Line; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mutagenesis, Insertional; Mutation

Bacterial type IV secretion systems are macromolecule transporters with essential functions for horizontal gene transfer and for symbiotic and pathogenic interactions with eukaryotic host cells. Helicobacter pylori, the causative agent of type B gastritis, peptic ulcers, gastric adenocarcinoma, and mucosa-associated lymphoid tissue (MALT) lymphoma, uses the Cag type IV secretion system to inject its effector protein CagA into gastric cells. This protein translocation results in altered host cell gene expression profiles and cytoskeletal rearrangements, and it has been linked to cancer development. Interactions of CagA with host cell proteins have been studied in great detail, but little is known about the molecular details of CagA recognition as a type IV secretion substrate or of the translocation process. Apart from components of the secretion apparatus, we previously identified several CagA translocation factors that are either required for or support CagA translocation. To identify protein-protein interactions between these translocation factors, we used a yeast two-hybrid approach comprising all cag pathogenicity island genes. Among several other interactions involving translocation factors, we found a strong interaction between the coupling protein homologue Cagβ (HP0524) and the Cag-specific translocation factor CagZ (HP0526). We show that CagZ has a stabilizing effect on Cagβ, and we demonstrate protein-protein interactions between the cytoplasmic part of Cagβ and CagA and between CagZ and Cagβ, using immunoprecipitation and pull-down assays. Together, our data suggest that these interactions represent a substrate-translocation factor complex at the bacterial cytoplasmic membrane.

Topics: Antigens, Bacterial; Bacterial Proteins; Bacterial Secretion Systems; Bacterial Translocation; Helicobacter Infections; Helicobacter pylori; Immunoblotting; Interleukin-8; Protein Interaction Domains and Motifs; Two-Hybrid System Techniques

Chronic inflammation associated with Helicobacter pylori infection is a risk factor of gastric adenocarcinoma. Interleukin-8 (IL-8) plays an important role in gastric mucosal inflammation induced by H. pylori infection. Recently, studies on the association of genetic polymorphisms of various proinflammatory cytokines with gastric carcinogenesis showed varying results on the basis of the ethnicity. We conducted this study to investigate the association of IL-8-251 A/T polymorphism with gastric carcinogenesis in H. pylori-infected Koreans.. The IL-8-251 A/T polymorphism was identified by polymerase chain reaction-restriction fragment length polymorphism using DNA from a total of 605 H. pylori-infected subjects; 206 controls, 149 chronic atrophic gastritis and/or intestinal metaplasia, 97 gastric dysplasia, and 153 gastric adenocarcinoma. Degrees of gastric mucosal inflammation and mucosal IL-8 level were also assessed.. The IL-8-251 A carriers showed a higher risk of gastric adenocarcinoma (adjusted odds ratio 2.06, 95% confidence interval 1.16-3.68) than IL-8-251 T/T genotypes. The IL-8-251 A allele was also significantly associated with the degree of neutrophil infiltration, atrophy, and intestinal metaplasia in a younger age group. Among the chronic atrophic gastritis and/or intestinal metaplasia group, mucosal IL-8 level was significantly higher in subjects with IL-8-251 A allele than those with IL-8-251 T/T genotypes (P=0.011).. The IL-8-251 A allele is associated with higher IL-8 production, more severe inflammation, mucosal atrophy, and intestinal metaplasia than IL-8-251 T/T genotype in H. pylori-infected hosts. The IL-8-251 A allele may also increase the risk of gastric adenocarcinoma through an enhanced inflammatory process in H. pylori-infected Koreans.

Topics: Adenocarcinoma; Age Factors; Analysis of Variance; Asian People; Female; Gastric Mucosa; Gastritis, Atrophic; Gene Frequency; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Korea; Male; Metaplasia; Middle Aged; Odds Ratio; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Sex Factors; Stomach Neoplasms

Helicobacter pylori infection is related to gastric cancer development, and chronic inflammation is presumed to be the main cause. The aim of the present study was to evaluate the influence of H. pylori cagA, vacA, iceA, and babA genotypes on COX-2, IL-1beta, and IL-8 expression.. Of the 217 patients included in the study, 26 were uninfected, 127 had chronic gastritis and were H. pylori-positive, and 64 had gastric cancer. Bacterial genotypes were evaluated by polymerase chain reaction (PCR), and the expression values were determined by quantitative real-time PCR and immunohistochemistry.. An association was found between the infection with cagA, vacA s1m1 strains and gastric cancer development. Regarding the 3' region of the cagA gene, we also found an association between the infection with cagA EPIYA-ABCCC strains and clinical outcome. Higher levels of IL-8, IL-1beta, and COX-2 were detected in gastric mucosa from infected patients with chronic gastritis, and they were also associated with the infection by cagA, vacA s1m1 strains. The IL-8 and IL-1beta levels decrease significantly from chronic gastritis to gastric cancer, while the relative expression remained unaltered when COX-2 expression was analyzed among patients with gastritis and cancer.. Since inflammatory response to H. pylori infection plays an important role in cellular proliferation and gastric mucosal damage, the up-regulation of IL-1beta, IL-8, and COX-2 in patients with chronic gastritis has an important clinical implication in gastric carcinogenesis.

Topics: Adult; Aged; Aged, 80 and over; Cyclooxygenase 2; Female; Gastritis; Gene Expression; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Stomach Neoplasms; Up-Regulation

The inflammatory response in Helicobacter pylori-infected gastric tissue is mediated by cag pathogenicity island (PAI)-dependent activation of nuclear factor-kappaB (NF-kappaB). Phosphatidylinositol 3-kinase (PI3K)/Akt signaling is known to play a role in NF-kappaB activation, but little information is available on the relationship between H. pylori and PI3K/Akt signaling in gastric epithelial cells. We examined whether H. pylori activates Akt in gastric epithelial cells, the role of cag PAI in this process and the role of Akt in regulating H. pylori-induced NF-kappaB activation.. Phosphorylated Akt was detected in epithelial cells of H. pylori-positive gastric tissues. Although Akt was activated in MKN45 and AGS cells by coculture with cag PAI-positive H. pylori strains, a cag PAI-negative mutant showed no activation of Akt. H. pylori also induced p65 phosphorylation. PI3K inhibitor suppressed H. pylori-induced p65 phosphorylation and NF-kappaB transactivation, as well as interleukin-8 expression. Furthermore, transfection with a dominant-negative Akt inhibited H. pylori-induced NF-kappaB transactivation. Transfection with small interference RNAs for p65 and Akt also inhibited H. pylori-induced interleukin-8 expression.. The results suggest that cag PAI-positive H. pylori activates Akt in gastric epithelial cells and this may contribute to H. pylori-mediated NF-kappaB activation associated with mucosal inflammation and carcinogenesis.

Topics: Cell Line; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; RNA Interference; Transcription Factor RelA

Matrix metalloproteinases (MMPs) are endopeptidases which perform important functions in extracellular matrix remodeling, cell proliferation, and inflammatory processes. Here, we compared MMP-3 levels with those of tissue inhibitor of metalloproteinases (TIMP)-1 and several inflammatory cytokines in gastric ulcer (GU) patients.. This study enrolled 50 patients with GU and 6 with functional dyspepsia (FD). Samples of gastric mucosa from the antrum and the ulcer site were harvested from GU patients and of antral mucosa alone from FD patients during upper gastrointestinal endoscopy. Mucosal biopsy tissues were cultured for 24 h, and the culture supernatant was measured for levels of MMP-3, TIMP-1, IL-1beta, IL-6, and IL-8.. All GU patients were positive for Helicobacter pylori, while all FD patients were negative. Antral levels of TIMP-1, IL-1beta, IL-6, and IL-8 were significantly higher in GU than FD patients. Further, MMP-3 levels were significantly higher in GU patients at the ulcer site than in the antrum, and had a significantly positive correlation with TIMP-1, IL-1beta, IL-6, and IL-8.. MMP-3 levels were significantly higher at the ulcer site than in the antrum, suggesting that MMP-3 may perform an important function in gastric ulcer healing.

Topics: Adult; Aged; Aged, 80 and over; Dyspepsia; Endoscopy, Gastrointestinal; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Linear Models; Male; Matrix Metalloproteinase 3; Middle Aged; Statistics, Nonparametric; Stomach Ulcer; Tissue Inhibitor of Metalloproteinase-1; Wound Healing

Helicobacter pylori is the most frequent cause of infection-induced cancer worldwide. Gastric carcino-genesis is the consequence of the important and life-long inflammation induced by H. pylori in the stomach. Gastric carcinogenesis, can be studied in many ways. In this chapter, we focus on some aspects concerning the bacteria, and others concerning the host. On the bacterial side, the methods exploring the presence of the cag pathogenicity island including cagA and the consequences on epithelial cells are presented. On the host side, tissue microarray, immunohistochemistry and chromogenic in situ hybridization (CISH) are described.

Topics: Amino Acid Sequence; Antigens, Bacterial; Bacterial Proteins; Bacterial Typing Techniques; Base Sequence; Cell Culture Techniques; Cells, Cultured; Coculture Techniques; Epithelial Cells; Gastric Mucosa; Genes, ras; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Molecular Sequence Data; Proto-Oncogene Proteins; Proto-Oncogene Proteins p21(ras); ras Proteins; Sequence Alignment; Stomach Neoplasms; Tissue Array Analysis

Nuclear factor-kappaB (NF-kappaB) plays a major role in host inflammatory responses and carcinogenesis and as such is an important drug target for adjuvant therapy. In this study, we examined the effect of caffeic acid phenethyl ester (CAPE), an NF-kappaB inhibitor, on Helicobacter pylori (H. pylori)-induced NF-kappaB activation in cell culture and chronic gastritis in Mongolian gerbils. In AGS gastric cancer cells, CAPE significantly inhibited H. pylori-stimulated NF-kappaB activation and mRNA expression of several inflammatory factors in a dose-dependent manner, and prevented degradation of IkappaB-alpha and phosphorylation of p65 subunit. To evaluate the effects of CAPE on H. pylori-induced gastritis, specific pathogen-free male, 6-week-old Mongolian gerbils were intragastrically inoculated with H. pylori, fed diets containing CAPE (0-0.1%) and sacrificed after 12 weeks. Infiltration of neutrophils and mononuclear cells and expression of NF-kappaB p50 subunit and phospho-IkappaB-alpha were significantly suppressed by 0.1% CAPE treatment in the antrum of H. pylori-infected gerbils. Labeling indices for 5'-bromo-2'-deoxyuridine both in the antrum and corpus and lengths of isolated pyloric glands were also markedly reduced at the highest dose, suggesting a preventive effect of CAPE on epithelial proliferation. Furthermore, in the pyloric mucosa, mRNA expression of inflammatory mediators including tumor necrosis factor-alpha, interferon-gamma, interleukin (IL)-2, IL-6, KC (IL-8 homologue), and inducible nitric oxide synthase was significantly reduced. These results suggest that CAPE has inhibitory effects on H. pylori-induced gastritis in Mongolian gerbils through the suppression of NF-kappaB activation, and may thus have potential for prevention and therapy of H. pylori-associated gastric disorders.

Topics: Animals; Blotting, Western; Caffeic Acids; Cytotoxins; Gastric Mucosa; Gastritis; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Immunoenzyme Techniques; Interferon-gamma; Interleukin-6; Interleukin-8; Male; NF-kappa B; Phenylethyl Alcohol; Phosphorylation; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

MicroRNAs (miRNAs) are small, noncoding RNAs that regulate gene expression at posttranscriptional level. H. pylori is a major human pathogenic bacterium in gastric mucosa. To date, the role of miRNAs in response to H. pylori infection has not been explored.. The expression profile of cellular miRNAs during H. pylori infection was analyzed by using microarray and quantitative reverse-transcriptase polymerase chain reaction. The potential target of miR-155 was identified by luciferase assay and Western blot. Promoter analysis and inhibitor experiment were used to investigate the pathway involved in the induction of miR-155. Examination of miR-155 function was performed by overexpression and inhibition of miR-155.. H. pylori was able to increase the miR-155 expression in gastric epithelial cell lines and gastric mucosal tissues, and nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) pathway were required for the induction of miR-155. miR-155 may down-regulate IkappaB kinase epsilon, Sma- and Mad-related protein 2 (SMAD2), and Fas-associated death domain protein. Furthermore, the overexpression of miR-155 negatively regulated the release of interleukin-8 and growth-related oncogene-alpha.. This study provides the first description of increased expression of miR-155 in H. pylori infection, and miR-155 may function as novel negative regulator that help to fine-tune the inflammation response of H. pylori infection.

Topics: Adult; Chemokine CXCL1; Fas-Associated Death Domain Protein; Female; Gastric Mucosa; Gene Expression Profiling; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Kinase; Inflammation; Interleukin-8; Male; MicroRNAs; Middle Aged; NF-kappa B; Oligonucleotide Array Sequence Analysis; Smad2 Protein

Helicobacter pylori CagA, translocated into gastric epithelial cells, induces IL-8 expression through the signalling pathways, including extracellular signal-regulated kinase (ERK) and nuclear factor-kappaB (NF-kappaB). We previously demonstrated that CagA interacts with host alphaPix. The present study was purposed to determine the role of the interaction of alphaPix with CagA on the signalling pathways for IL-8 expression in H. pylori-infected gastric epithelial cells.. H. pylori HP99 strain (CagA+, VacA+) was infected to gastric epithelial AGS cells transfected with non-targeting (NT) or alphaPix- targeting siRNA. Activation of signalling molecules including p21-activated kinase (PAK), ERK and NF-kappaB, and expression of IL-8 in the cells were assessed.. H. pylori CagA was delivered into AGS cells and then interacted with alphaPix at 4 h following H. pylori infection. PAK1, ERK and NF-kappaB were activated in the cells containing NT and alphaPix siRNA at 1-2 h following H. pylori infection. However, after 4 h, the time when CagA was delivered into the cells, the activations of PAK1, ERK and NF-kappaB were inhibited by down-regulation of alphaPix using siRNA but not by NT siRNA. The results indicate that alphaPix is required for H. pylori-mediated signalling of PAK1, ERK and NF-kappaB. Additionally, alphaPix siRNA suppressed IL-8 induction after translocation of CagA into the cells, indicating that interaction of CagA with alphaPix is critical for CagA-mediating signalling for IL-8 expression.. The interaction of alphaPix with CagA activates PAK1, ERK and NF-kappaB, which induces IL-8 expression in H. pylori-infected gastric epithelial cells.

Topics: Antigens, Bacterial; Bacterial Proteins; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gastric Mucosa; Guanine Nucleotide Exchange Factors; Helicobacter Infections; Humans; Interleukin-8; NF-kappa B; p21-Activated Kinases; Rho Guanine Nucleotide Exchange Factors; Signal Transduction

We examined the kinetics of IL-8 production and CagA status in AGS infected with Helicobacter pylori, 26695 (Western CagA-type), HPK5 (Eastern CagA-type) and isogenic cagA-disrupted mutants, exposed to different pH (pH 6 and pH 3). IL-8 was produced in the early and late phases after infection in CagA-dependent and -independent manners, respectively, irrespective of CagA-type. The wild-type exposed to low pH tremendously reduced IL-8 level at early phase, but restored with urea, suggesting that low pH exerted the kinetics of H. pylori-induced IL-8 production in CagA-dependent manner and urea was necessary for effective induction. CagA and phosphorylated CagA increased time-dependently after infection. Phosphorylated CagA from 26695, but not HPK5, rapidly peaked, consistent with the kinetics of IL-8 induction and appearance of hummingbird phenotype. ATF3 transcripts peaked late phase by wild-type, however, induced in two peaks early and late phases by cagA-disrupted mutants, indicating that different CagA-type proteins altered ATF3 induction in the infected cells.

Topics: Activating Transcription Factor 3; Antigens, Bacterial; Bacterial Proteins; Cell Line, Tumor; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Interleukin-8; Phosphorylation

Relations between host genetic factors and clinical outcomes of Helicobacter pylori infection are variable among ethnicities. The aim of this study was to examine gastric mucosal cytokines, matrix metalloproteinase 3 (MMP-3), and serum pepsinogen levels before and after eradication of H. pylori according to IL-1B genotypes and benign gastroduodenal phenotypes in a Korean population.. A total of 349 Koreans including H. pylori-infected subjects (n=230) and H. pylori-negative controls (n=119) were enrolled. The former subjects were classified into groups according to the presence of non-atrophic gastritis (n=74), atrophic gastritis (n=56), gastric ulcer (n=37), and duodenal ulcer (n=63). IL-1B polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Gastric mucosal IL-1beta, IL-8, and MMP-3, and serum pepsinogen I and II levels were measured by ELISA and radioimmunoassay, respectively.. There were no significant differences between the IL-1B-31/-511 haplotype (TT/CC, CT/CT, and CC/TT) frequencies among the H. pylori-positive and -negative groups. The genotypes of IL-1B-31/-511 polymorphisms did not affect clinical phenotypes, inflammatory cytokines, MMP-3, and pepsinogen secretion. Subjects with H. pylori-infected atrophic gastritis exhibited significantly higher basal levels of cytokines and a lower pepsinogen I/II ratio than those of other groups. Following H. pylori eradication, inflammatory cytokines significantly decreased and the pepsinogen I/II ratio increased in all groups.. Mucosal inflammatory cytokines, MMP-3, and pepsinogen secretion are related to gastroduodenal phenotypes but not to IL-1B genotypes. Eradication of H. pylori can reduce mucosal inflammation and restore pepsinogen secretion.

Topics: Adult; Case-Control Studies; Female; Gastric Mucosa; Haplotypes; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1beta; Interleukin-8; Korea; Male; Matrix Metalloproteinase 3; Middle Aged; Pepsinogen A; Phenotype; Polymorphism, Genetic

Helicobacter pylori infection is the most common cause of gastritis, gastric ulcer and adenocarcinoma. It has proven difficult to cure because of its capability to develop strains resistant to antibiotics. The effect of three strains of lactic acid bacteria (LAB) and bovine colostral preparations on the adhesion of H. pylori NCTC 11637 on gastric adenocarcinoma (AGS) cells and on the interleukin (IL)-8 production was studied. Before infection, H. pylori were pretreated with Lactobacillus plantarum MLBPL1, Lactobacillus rhamnosus GG, Lactococcus lactis, or with a colostral preparation with or without specific H. pylori antibodies. The relative number of H. pylori adhered on AGS cells was determined by urease test. IL-8 produced by the cells was studied by enzyme-linked immunosorbent assay. Colostral preparations with and without specific antibodies reduced the adhesion of H. pylori on AGS cells in a dose-dependent manner. Live LAB at a concentration of 10(10) CFU/ml reduced the adhesion by approximately 50% (P < 0.05). After the infection of AGS cells by H. pylori, the IL-8 level rose up to about 10-fold (5500 +/- 1600 pg/ml). Pretreatment of H. pylori with colostral preparations or high concentrations of LAB prevented this IL-8 rise. Similar effect was seen with live and heat-killed LAB, the live LAB being more effective. Heat-killed LAB at a concentration of 10(10) CFU/ml rose the IL-8 level of non-infected cells significantly. Suppression of IL-8 production by LAB or colostral products could have a suppressive effect on inflammation in Helicobacter infection.

Topics: Adenocarcinoma; Animals; Antibodies; Bacterial Adhesion; Biological Products; Cattle; Cell Line, Tumor; Colostrum; Culture Media, Conditioned; Dose-Response Relationship, Drug; Dose-Response Relationship, Immunologic; Female; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Lactobacillus; Pregnancy; Probiotics

To investigate the effect of Lactobacillus bulgaricus (LBG) on the Toll-like receptor 4 (TLR4) pathway and interleukin-8 (IL-8) production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide (H pyloriSS1-LPS).. SGC-7901 cells were treated with H pyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth (LBG-(s)). Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor beta-activated kinase 1 (TAK1), anti-phospho-TAK1, anti-nuclear factor kappaB (NF-kappaB), anti-p38 mitogen-activated protein kinase (p38MAPK), and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA.. H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAK1, subsequently enhanced the activation of NF-kappaB and the phosphorylation of p38MAPK in a time-dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-(s) pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-kappaB, and consequently blocked IL-8 production.. H pyloriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-(s) prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.

Topics: Adenocarcinoma; Cell Line, Tumor; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillus; Lipopolysaccharides; Probiotics; Stomach Neoplasms; Toll-Like Receptor 4

The chronic inflammatory process in patients with Crohn's disease (CD) may affect any part of the gastrointestinal (GI) tract. The pathogenesis of CD involves immunological abnormalities, including deficient or excessive expression of cytokines. We examined Helicobacter pylori infection status, endoscopic and histopathological findings, and cytokine production in the duodenum of CD patients in comparison with controls.. Thirty-eight CD patients underwent diagnostic upper GI endoscopy. Twelve age- and sex-matched health checkup examinees were used as controls. H. pylori infection status was assessed by the (13)C-urea breath test. At the time of endoscopy, two biopsy specimens each were obtained from the second portion of the duodenum, one for hematoxylin-eosin staining and immunohistochemical analysis with anti-CD68 antibody, and one for in vitro organ culture. Interleukin (IL)-6 and -8 levels were measured in organ culture supernatant by enzyme-linked immunosorbent assay.. H. pylori infection was significantly (P<0.05) more frequent in controls (42%) than in CD patients (8%). In the duodenum, erosions or ulcers were more frequent in CD patients (53%) than in controls (8%). Mononuclear cell infiltration in the duodenum was more severe in CD patients than in controls and IL-6 production was higher, whereas IL-8 production showed no significant difference. CD68+ cells in the duodenum were more prominent in CD patients than in controls.. H. pylori infection is unlikely in CD patients, but they show immunological abnormalities in the duodenum, possibly from innate immune responses.

Topics: Adolescent; Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biopsy; Breath Tests; Case-Control Studies; Crohn Disease; Duodenal Ulcer; Duodenoscopy; Duodenum; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Innate; Interleukin-6; Interleukin-8; Organ Culture Techniques; Urea; Young Adult

Induction of inducible nitric oxide synthase (iNOS) may be involved in carcinogenesis of the stomach, because nitric oxide (NO) derived from iNOS can exert DNA damage and post-transcriptional modification of target proteins. In the present study, we investigated the correlation between endoscopic findings and iNOS mRNA expression/NO-modified proteins in the gastric mucosa.. Fifty patients were prospectively selected from subjects who underwent upper gastrointestinal chromoendoscopy screening for abdominal complaints. The Helicobacter pylori (H. pylori) status of patients was determined by anti-H. pylori IgG antibody levels. We classified the mucosal area of the fundus as F0, fine small granules; F1, edematous large granules without a sulcus between granules; F2, reduced-size granules with a sulcus between granules; and F3, irregular-sized granules with extended sulcus between granules. Gastritis was graded using the visual analog scale of the Updated Sydney System. The expression of interleukin (IL)-8 and iNOS mRNA was assayed in gastric biopsy specimens by reverse transcription-polymerase chain reaction. NO-modified proteins were analyzed by Western blotting using novel monoclonal antibodies against nitrotyrosine.. A total of 91.7% (11/12) of the F0 group was H. pylori-negative, whereas 94.7% (36/38) of the F1-3 groups was H. pylori-positive. Spearman's analysis showed good correlation between the endoscopic grading and the score of chronic inflammation (r=0.764) and glandular atrophy (r=0.751). The expression of IL-8 mRNA was significantly increased in F1, F2, and F3 cases compared with the F0 group, with no significant differences among them. iNOS mRNA was significantly increased in the F3 group compared with the other groups, with increased nitration of tyrosine residues of proteins.. The proposed classification by chromoendoscopy is useful for screening patients for atrophic and iNOS-expressing gastric mucosa with NO-modified proteins in H. pylori-associated atrophic gastric mucosa.

Topics: Antibodies, Bacterial; Atrophy; Biomarkers; Gastric Mucosa; Gastritis, Atrophic; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Nitric Oxide; Nitric Oxide Synthase Type II; Prospective Studies; Proteins; RNA, Messenger; Severity of Illness Index; Tyrosine

Topics: Demography; Duodenal Ulcer; Female; Gastric Mucosa; Gene Expression Regulation; Genetic Predisposition to Disease; Haplotypes; Helicobacter Infections; Helicobacter pylori; Humans; India; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Polymorphism, Genetic; RNA, Messenger; Tumor Necrosis Factor-alpha

The plasticity region of the Helicobacter pylori genome comprises strain-specific gene loci. We performed genotyping and functional biology analysis of one such locus (jhp940) that was previously found to be functionally unknown but present in gastric cancer-associated strains from many different countries. We found its geographic prevalence to be independent of cagA presence and disease status. Cloning, expression, and purification of JHP940 revealed a novel, approximately 36-kDa protein in a biologically active form which elicited strong and significant levels of tumor necrosis factor alpha and interleukin-8 in human macrophages. Also, JHP940 was able to induce enhanced translocation of the transcription factor NF-kappaB complex in cultured macrophages. The induction of the proinflammatory cytokines by JHP940, therefore, points to its putative role in chronic gastric inflammation and, possibly, the various other outcomes of H. pylori infection, including gastric cancer.

Topics: Antigens, Bacterial; Bacterial Proteins; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Macrophages; NF-kappa B; Tumor Necrosis Factor-alpha

Genetic variations of the inflammatory IL-8 and TNF-alpha genes can influence the outcome of gastric alterations. Our aims were to determine the prevalence and effect of the T-251A functional polymorphism of IL-8 and the G-308A polymorphism of TNF-alpha in histological and macroscopic gastric diseases related to Helicobacter pylori infection.. Genomic DNA was extracted from biopsy samples from patients with gastritis (n=86, H. pylori positive=41), atrophy (n=32, H. pylori positive=13), intestinal metaplasia (IM) (n=43, H. pylori positive=22) and from histologically negative patients (n=57). The samples were divided by macroscopic diagnosis into erosion and negative groups. The T-251A polymorphism was examined with the amplification refractory mutation system method; the G-308A polymorphism was determined by the polymerase chain reaction-restriction fragment length polymorphism method. For statistical evaluation, Fischer's exact test was used.. In the case of T-251A of IL-8, the frequency of the A/A genotype was significantly increased in gastritis (P=0.049) and IM (P=0.038) groups as compared with the histologically negative ones. No relationship was found between macroscopic erosions and H. pylori infection. In the case of G-308A, the G/G genotype frequency was statistically increased in erosions as compared with negative groups (P=0.035). No difference in the distribution of G-308A genotypes in relation to histological alterations and the H. pylori infection was observed.. The effect of the polymorphism of IL-8 seems to be relevant in the pathogenesis of histological gastritis and IM, and the effect of the polymorphism of TNF-alpha is relevant in the pathogenesis of macroscopic erosive gastritis.

Topics: Atrophy; Gastritis; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Metaplasia; Polymorphism, Genetic; Stomach; Tumor Necrosis Factor-alpha

In Helicobacter pylori gastritis gastric epithelium plays a central role in the innate immunity to H. pylori. However, epithelial receptors interacting with H. pylori have been poorly characterized so far. Recently a new triggering receptor expressed on myeloid cells-1 (TREM-1) has been identified on human neutrophils and monocytes. On these cells TREM-1 triggers innate immunity by stimulating the secretion of interleukin (IL)-8 and tumour necrosis factor (TNF)-alpha and thus amplifies bacterial-induced inflammation. In this study expression and function of TREM-1 in gastric epithelium exposed to H. pylori has been investigated. TREM-1 mRNA and protein were expressed on gastric epithelial cell lines as demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence activated cell sorter analysis. Gastric epithelial TREM-1 expression was up-regulated directly by H. pylori and was independent of epithelial IL-8 induced by H. pylori. Immunohistochemistry and tissue RT-PCR demonstrated significantly stronger TREM-1 expression in H. pylori gastritis compared with the non-inflamed gastric mucosa supporting in vivo that epithelial TREM-1 is up-regulated during H. pylori infection. Stimulation of gastric epithelial TREM-1 receptor resulted in IL-8 up-regulation on mRNA and protein level, as shown by real-time PCR and immunoassay. This is the first study localizing TREM-1 on gastric epithelium. Functional data suggest that TREM-1 expressed on gastric epithelium amplifies inflammation of the underlying gastric mucosa by up-regulation of IL-8.

Topics: Cell Line; Epithelial Cells; Gastric Mucosa; Gastritis; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Innate; Interleukin-8; Lipopolysaccharides; Membrane Glycoproteins; Receptors, Immunologic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Triggering Receptor Expressed on Myeloid Cells-1; Up-Regulation

Although probiotics have been reported to reduce the gastric inflammatory response to Helicobacter pylori infection, little information is available regarding the molecular mechanisms behind this reduction. This study investigates the role of conjugated linoleic acids (CLA) produced by probiotics in interactions of IkappaB kinase (IKK) and heat shock protein 90 (Hsp90) to activate the nuclear factor-kappaB (NF-kappaB) signaling pathway in human gastric epithelial cells infected with H. pylori. Conditioned medium (CM) containing Lactobacillus acidophilus-producing CLA significantly inhibited the activated NF-kappaB signals and the upregulated expression of interleukin-8 (IL-8) in MKN-45 cells infected with H. pylori. Pretreatment with CM with CLA attenuated the increased IKK activity induced by H. pylori. Transfection of siRNA for IKK-beta dramatically reduced H. pylori-induced IkappaBalpha phosphorylation, but siRNA for IKK-alpha had little effect on IkappaBalpha phosphorylation, although the siRNA for IKK-alpha significantly decreased IL-8 production. Furthermore, Hsp90 was associated with IKK-alpha and IKK-gamma in H. pylori-infected cells, and CM with CLA dissociated the complex between Hsp90 and IKK-gamma. These results suggest that CLA produced by probiotics has anti-inflammatory activity in gastric epithelial cells infected with H. pylori via dissociation of the IKK-gamma and Hsp90 complex.

Topics: Cell Line; Enzyme Activation; Gastric Mucosa; Helicobacter Infections; HSP90 Heat-Shock Proteins; Humans; I-kappa B Kinase; Interleukin-8; Lactobacillus; Linoleic Acids, Conjugated; NF-kappa B; Phosphorylation; RNA, Small Interfering; Signal Transduction

Arachidonic acid metabolites have been considered as pivotal mediators in Helicobacter pylori-induced inflammatory response, which are mainly metabolized by two distinct enzymes: cyclooxygenase (COX) and lipoxygenase (LOX). While COX has become well known to play a key role in either carcinogenesis or inflammation related to H. pylori infection, little is known regarding the implication of LOX in H. pylori infection. In this study, we evaluated the roles of 5-LOX and its metabolites in H. pylori-induced host responses and further a potential beneficial action of specific LOX inhibitors against H. pylori infection.. Expressions of cytosolic phospholipase A(2) (cPLA(2)), COX-2, and 5-LOX after H. pylori infection were evaluated by immunofluorescence staining and Western blotting. Synthesis of LOX metabolites was measured with reversed-phase high-performance liquid chromatography. For analyzing the influence of 5-LOX inhibitors, nordihydroguaiaretic acid (NDGA) and geraniin, on H. pylori-induced inflammatory responses, RNase protection assay and RT-PCR were performed.. H. pylori stimulated the translocation of cPLA(2) from cytoplasm to nucleus and increased the biosynthesis of hydroxyeicosatetraenoic acids (HETEs) as a predominant form of 5S-HETE in gastric epithelium. NDGA exerted a strong suppression activity of H. pylori-induced 5-LOX signaling. The administration of LOX inhibitors was related with down-expression of proinflammatory mediators such as interleukin-8 and tumor necrosis factor-alpha in both H. pylori-infected gastric epithelial cells and macrophage cells.. LOX modulation with its specific inhibitors could impose significant anti-inflammatory responses after H. pylori infection, based on the fact that H. pylori infection provoked gastric inflammation through metabolizing arachidonic acid by the 5-LOX pathway.

Topics: Animals; Arachidonate 5-Lipoxygenase; Arachidonic Acid; Cell Line; Down-Regulation; Epithelial Cells; Glucosides; Helicobacter Infections; Helicobacter pylori; Humans; Hydrolyzable Tannins; Interleukin-8; Lipoxygenase Inhibitors; Macrophages; Masoprocol; Mitogen-Activated Protein Kinase 3; NF-kappa B; Oxidation-Reduction; Signal Transduction; Tumor Necrosis Factor-alpha

Intracellular pathogen receptor NOD1 is involved in the epithelial cell sensing Helicobacter pylori, which results in a considerable interleukin (IL)-8 production. The aim of this study was to evaluate the relationship between NOD1 and IL-8 genetic polymorphisms and the development of H. pylori-induced gastritis and duodenal ulcer (DU), as compared with TLR4 polymorphisms.. Eighty-five patients with DU and 135 patients with gastritis were enrolled in the study. Seventy-five serologically H. pylori-positive subjects without gastric or duodenal symptoms served as controls. The G796A (E266K) NOD1 polymorphism was determined by restriction fragment length polymorphism, and the -251 IL-8 polymorphism by amplification refractory mutation system method. The TLR4 (ASP/299/Gly and Thr/399/Ile) gene polymorphisms were examined by melting point analysis.. AA homozygote mutant variants of NOD1 were detected in 20% of the H. pylori-positive patients with DU versus 7% of H. pylori-positive patients with gastritis and versus 6% of the H. pylori-positive healthy controls. The IL-8 heterozygote mutant variant was detected with a significantly higher frequency among the DU patients and those with gastritis than among the H. pylori-positive controls. However, no significant correlation concerning the frequency of the TLR4 gene polymorphism could be revealed between any group of patients and the controls.. E266K CARD4/NOD1, but not the TLR4 gene polymorphism increases the risk of peptic ulceration in H. pylori-positive patients. The -251 IL-8 polymorphism was significantly associated with either gastritis or DU in H. pylori-infected subjects. Host factors including intracellular pathogen receptors and IL-8 production play an important role in H. pylori-induced gastric mucosal damage.

Topics: Case-Control Studies; Duodenal Ulcer; Gastritis; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Nod1 Signaling Adaptor Protein; Polymorphism, Genetic; Toll-Like Receptor 4

To study whether H pylori are associated with chronic cholecystitis.. The subjects were divided into three groups: H pylori-infected cholecystitis group, H pylori-negative cholecystitis group and control group. Pathologic changes of the gallbladder were observed by optic and electronic microscopes and the levels of interleukin-1, 6 and 8 (IL-1, 6 and 8) were detected by radioimmunoassay.. Histological evidence of chronic cholecystitis including degeneration, necrosis, inflammatory cell infiltration, were found in the region where H pylori colonized. Levels of IL-1, 6 and 8 in gallbladder mucosa homogenates were significantly higher in H pylori-infected cholecystitis group than those in H pylori-negative cholecystitis group and control group.. H pylori infection may be related to cholecystitis.

Topics: Case-Control Studies; Cholecystitis; Chronic Disease; Gallbladder; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Metaplasia; Mucous Membrane

Helicobacter pylori CagA is translocated into gastric epithelial cells by a type IV secretion system and interacts with the Src homology 2 phosphatase, altering cell morphology. Multiple EPIYA motifs in CagA are associated with increased activity in cells and with gastric cancer. The aim of this work was to study the heterogeneity in activity in cells of multiple H. pylori single colonies isolated from a single patient and its association with polymorphism in cagA. The presence of cagA, cagE, cagT, and cag10 was studied with 318 H. pylori isolates from the antra and corpora of 18 patients. AGS gastric epithelial cells were infected with 75 isolates, and interleukin-8 (IL-8) secretion, cytoskeletal changes, CagA translocation, and tyrosine phosphorylation were measured. The cagA 3'-variable region was sequenced for 30 isolates to determine the number and types of EPIYA motifs. Isolates from an individual stomach were usually genetically related and had quantitatively similar phenotypic effects on cells (IL-8 induction and cytoskeletal changes). However, strains from different patients with similar CagA EPIYA motif patterns varied widely in these phenotypes. Among isolates with an EPIYA-ABC pattern, the phenotype was variable: IL-8 induction ranged from 200 to 1,200 pg/ml, and morphological changes occurred in 20 to 70% of cells. In several cases, cagA sequence diversity appeared to explain the lack of CagA activity, as isolates with an EPIYA-ACC pattern or a modified B motif had reduced cell activity. cag pathogenicity island-positive H. pylori isolates displayed a high level of heterogeneity in the capacity to induce IL-8 secretion and morphological changes; an absent or modified B motif was associated with low activity.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Amino Acid Sequence; Antigens, Bacterial; Bacterial Proteins; Cell Line; Child; Epithelial Cells; Female; Genetic Variation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Mexico; Middle Aged; Molecular Sequence Data; Phenotype; Sequence Analysis, DNA; Stomach

The intensity of the inflammation induced by Helicobacter pylori colonization is associated with the development of distal gastric cancer (GC). The host response to H. pylori has been related to genetic polymorphisms that influence both innate and adaptive immune responses.Our aim was to investigate whether the presence of the TLR4 Asp299Gly, TLR4 Thr399Ile and IL-8-251 A/T polymorphisms had any influence in the development of distal GC in a Mexican population.. We studied 337 patients that were divided in two groups: 78 patients with histologically confirmed distal GC and 259 non-cancer controls. The presence of H. pylori in the control population was defined by positive results of at least two of four diagnostic tests: serology, histology, rapid urease test and culture. Human DNA was purified and genotyped for TLR4 Asp299Gly polymorphism by pyrosequencing, for TLR4 Thr399Ile by PCR-RFLP and for IL8-251 by the amplification refractory mutation system (ARMS)-PCR.. The non-cancer control group was found to be in Hardy-Weinberg equilibrium at the polymorphic loci studied (chi-square H-W = 0.58 for IL8-251, 0.42 for TLR4 Asp299Gly and 0.17 for TLR4 Thr399Ile). The frequencies of mutated alleles (homozygous plus heterozygous) were compared between cases and controls. We found no significant difference for TLR4- Asp299Gly [the 7.7% of distal GC patients and 7.7 % non-cancer controls (p = 0.82)] and for TLR4 Thr399Ile [the 1.3% of GC patients and the 5% of the control population (p = 0.2)]. In contrast, for IL-8-251 A/T, 80.77% of the GC patients and 66.4% in the control group age and gender matched had at least one copy of mutated allele (OR = 2.12, 95% CI = 1.1-4.2) (p = 0.023).. This study showed that the IL8-251*A allele could be related to the development of distal gastric cancer in this Mexican population.

Topics: Adult; Age Distribution; Aged; Aged, 80 and over; Case-Control Studies; Confidence Intervals; DNA, Bacterial; Female; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Incidence; Interleukin-8; Male; Mexico; Middle Aged; Odds Ratio; Polymorphism, Genetic; Probability; Reverse Transcriptase Polymerase Chain Reaction; Risk Assessment; RNA, Transfer, Asp; Sex Distribution; Stomach Neoplasms; Toll-Like Receptor 4

To analyze the status of expression of inflammation markers, antioxidant and oxidant enzymes in biopsies from patients diagnosed with gastritis, gastric ulcer (GU) and gastric cancer (GC) and the Helicobacter pylori virulence from these isolated biopsies in order to evaluate a possible association among these factors.. H. pylori genotype from isolated biopsies was performed by PCR. The pattern of expression of inflammation (TNF-alpha, IL-1beta, IL-8, IL-10 and IL-12), oxidant (iNOS and Nox1) and antioxidant markers (MnSOD, GPX and CAT) of biopsies from gastritis, GU, GC and control groups was performed by RT-PCR.. Different from other gastric diseases studied here, gastritis is characterized by an oxidative stress with significant expression of TNF-alpha, IL-8, IL-12, iNOS and Nox and significant absence of MnSOD and GPX expression. Gastritis was the only condition where there was an association between TNF-alpha or IL-8 expression and H. pylori cagA+/vacAs1 genotype. In this case, TNF-alpha expression was about 3 times higher when compared to control subjects.. In this study, only gastritis was found to be associated with significant oxidative stress marker expression of TNF-alpha and IL-8 that was also related to H. pylori virulence, suggesting that they are the main oxidant stress markers responsible to trigger an increase in ROS level that contributes to decrease the expression of the MnSOD and GPX.

Topics: Antioxidants; Gastritis; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-12; Interleukin-8; Multienzyme Complexes; NADH, NADPH Oxidoreductases; Nitric Oxide Synthase Type II; Oxidative Stress; Stomach Diseases; Stomach Neoplasms; Stomach Ulcer; Tumor Necrosis Factor-alpha; Virulence

To explore the interactions between the host, environment and bacterium responsible for the different manifestations of Helicobacter pylori infection, we examined the effect of acidic conditions on H. pylori-induced interleukin (IL)-8 expression. AGS gastric epithelial cells were exposed to acidic pH and infected with H. pylori[wild-type strain, its isogenic cag pathogenicity island (PAI) mutant or its oipA mutant]. Exposure of AGS cells to acidic pH alone did not enhance IL-8 production. However, following exposure to acidic conditions, H. pylori infection resulted in marked enhancement of IL-8 production which was independent of the presence of the cag PAI and OipA, indicating that H. pylori and acidic conditions act synergistically to induce gastric mucosal IL-8 production. In neutral pH environments H. pylori-induced IL-8 induction involved the NF-kappaB pathways, the extracellular signal-regulated kinase (ERK)-->c-Fos/c-Jun-->activating protein (AP-1) pathways, JNK-->c-Jun-->AP-1 pathways and the p38 pathways. At acidic pH H. pylori-induced augmentation of IL-8 production involved markedly upregulated the NF-kappaB pathways and the ERK-->c-Fos-->AP-1 pathways. In contrast, activation of the JNK-->c-Jun-->AP-1 pathways and p38 pathways were pH independent. These results might explain the clinical studies in which patients with duodenal ulcers had higher levels of IL-8 in the antral gastric mucosa than patients with simple H. pylori gastritis.

Topics: Cell Line; Duodenal Ulcer; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Interleukin-8; Signal Transduction

The ulcer-causing pathogen Helicobacter pylori uses directed motility, or chemotaxis, to both colonize the stomach and promote disease development. Previous work showed that mutants lacking the TlpB chemoreceptor, one of the receptors predicted to drive chemotaxis, led to less inflammation in the gerbil stomach than did the wild type. Here we expanded these findings and examined the effects on inflammation of completely nonchemotactic mutants and mutants lacking each chemoreceptor. Of note, all mutants colonized mice to the same levels as did wild-type H. pylori. Infection by completely nonchemotactic mutants (cheW or cheY) resulted in significantly less inflammation after both 3 and 6 months of infection. Mutants lacking either the TlpA or TlpB H. pylori chemotaxis receptors also had alterations in inflammation severity, while mutants lacking either of the other two chemoreceptors (TlpC and HylB) behaved like the wild type. Fully nonchemotactic and chemoreceptor mutants adhered to cultured gastric epithelial cells and caused cellular release of the chemokine interleukin-8 in vitro similar to the release caused by the wild type. The situation appeared to be different in the stomach. Using silver-stained histological sections, we found that nonchemotactic cheY or cheW mutants were less likely than the wild type to be intimately associated with the cells of the gastric mucosa, although there was not a strict correlation between intimate association and inflammation. Because others have shown that in vivo adherence promotes inflammation, we propose a model in which H. pylori uses chemotaxis to guide it to a productive interaction with the stomach epithelium.

Topics: Animals; Bacterial Adhesion; Bacterial Proteins; Cell Line; Chemotaxis; Epithelium; Female; Gastric Mucosa; Gastritis; Gene Deletion; Helicobacter Infections; Helicobacter pylori; Histocytochemistry; Humans; Interleukin-8; Membrane Proteins; Methyl-Accepting Chemotaxis Proteins; Mice; Mutagenesis, Insertional

Helicobacter pylori is an important pathogen in major gastroduodenal diseases, including inflammation with ulceration and gastric malignancies. Alterations in H. pylori associated cell turnover in gastric epithelial cells are examined in relation to inflammatory activity, bacteria load and cytokines which may improve knowledge concerning the outcome of gastric diseases caused by H. pylori. Antral biopsies from 42 dyspeptic patients including 27 H. pylori-positive and 15 H. pylori-negative patients were tested for apoptotic activity by the TUNEL assay, and immuno-histochemically for p53 and the proliferative marker Ki-67. H. pylori infection, bacteria load and inflammatory activity were associated with increased cell turnover as judged by enhanced activities of TUNEL, p53 and Ki-67. Only p53 was significantly correlated to IFN-gamma, IL-8 and IL-10. The H. pylori-positive state was furthermore accompanied by varying degrees of altered distribution pattern of the markers studied, with occasional presence of apoptosis in the deeper pit zones, upward extension of Ki-67 and to a lesser degree of p53. Given a similar pattern of change in proliferation and apoptosis in some neoplastic lesions in other parts of the gastrointestinal tract, such studies in cell turnover may provide insights valuable in the investigations of potential precursors of gastric malignancies.

Topics: Apoptosis; Biopsy; Cell Proliferation; Epithelial Cells; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Interferon-gamma; Interleukin-10; Interleukin-8; Ki-67 Antigen; Kinetics; Male; Pyloric Antrum; Tumor Suppressor Protein p53

H. pylori infection accounts for most cases of gastric cancer, but the initiating events remain unclear. The principal H. pylori pathogenicity-associated CagA protein disrupts intracellular SHP-2 signalling pathways including those used by the IL-6 family cytokines, IL-6 and IL-11. Imbalanced IL-6 family cytokine signalling in the gp130(757FF) mouse model of gastric cancer arising from hyperactivation of oncogenic STAT3 after altered SHP-2 : ERK1/2 signalling produces dysplastic antral tumours preceded by gastritis and metaplasia. In a cohort of patient gastric biopsies with known H. pylori and CagA status, we investigated whether (i) STAT3 and ERK1/2 activation is altered in H. pylori-dependent gastritis; (ii) these profiles are more pronounced in CagA+ H. pylori infection; and (iii) the expression of pro-inflammatory cytokines that activate STAT3 and ERK 1/2 pathways is associated with progression to gastric cancer. IL-6, IL-11, and activated STAT3 and ERK1/2 were quantified in antral biopsies from gastritic stomach, metaplastic tissue, and resected gastric cancer tissues. We observed significantly increased STAT3 and ERK1/2 activation (p = 0.001) in H. pylori-dependent gastritis, which was further enhanced in the presence of CagA+ H. pylori strains. Of known gastric ligands that drive STAT3 activation, IL-6 expression was increased after H. pylori infection and both IL-6 and IL-11 were strongly up-regulated in the gastric cancer biopsies. This suggests a mechanism by which IL-11 drives STAT3 activation and proliferation during gastric cancer progression. We addressed this using an in vitro approach, demonstrating that recombinant human IL-11 activates STAT3 and concomitantly increases proliferation of MKN28 gastric epithelial cells. In summary, we show increased STAT3 and ERK1/2 activation in H. pylori-dependent gastritis that is likely driven in an IL-6-dependent fashion. IL-11 expression is associated with adenocarcinoma development, but not gastritic lesions, and we identify a novel mechanism for IL-11 as a potent inducer of proliferation in the human gastric cancer setting.

Topics: Adenocarcinoma; Antigens, Bacterial; Bacterial Proteins; Biopsy; Cell Proliferation; Disease Progression; Enzyme Activation; Gastric Mucosa; Gastritis; Gene Expression Regulation, Neoplastic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-11; Interleukin-6; Interleukin-8; Mitogen-Activated Protein Kinase 3; Neoplasm Proteins; Proton Pump Inhibitors; Pyloric Antrum; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; STAT3 Transcription Factor; Stomach Neoplasms; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins

Capsaicin, the main pungent ingredient of hot red and chilli pepper, has been considered as not only a cytoprotective but also a detrimental agent to the gastric mucosa. However, the effect and mechanism of capsaicin that modulate the induction of pro-inflammatory cytokine in Helicobacter pylori-infected epithelial cells have not been investigated previously. Herein, we demonstrated that capsaicin inhibited the release of pro-inflammatory cytokine, interleukin-8 (IL-8) by H. pylori-infected gastric epithelial cells through nuclear factor-kappaB (NF-kappaB) signal pathway.. AGS or MKN45 cells as gastric epithelial cells and Vac A+, CagA+ wild-type H. pylori strain ATCC 49503 were used. Gastric epithelial cells were pre-treated with various concentrations of capsaicin and infected with H. pylori for different periods of time to determine IL-8 concentrations in culture supernatant by an ELISA assay. We measured IL-8 mRNA transcripts in H. pylori-infected gastric epithelial cells co-treated with capsaicin by reverse transcriptase-polymerase chain reaction analysis. We performed electrophoretic mobility shift assay to examine the NF-kappaB DNA binding activity with capsaicin and immunofluorescence microscopy to examine nuclear staining of p65. We also performed immunoblotting for IkappaB, IKK activity with capsaicin.. Capsaicin inhibits H. pylori-induced IL-8 production by gastric epithelial cells in dose- and time-dependent manner. Capsaicin as low as 100 micromol/L significantly inhibited IL-8 production in H. pylori-infected MKN45 cells (43.2% of control) at 24 hours incubation, whereas inhibited IL-8 production in H. pylori-infected AGS cells (70% of control). We confirmed that capsaicin inhibited IL-8 mRNA expression after infection of gastric epithelial cells with H. pylori for 6 hours. The addition of capsaicin (100 micromol/L) suppressed H. pylori-induced NF-kappaB activation in gastric epithelial cells at 1 hour post-infection. We also found that the degradation of IkappaB and IKK activation were inhibited by capsaicin.. Nontoxic dose of capsaicin inhibited H. pylori-induced IL-8 production by gastric epithelial cells through the modulation of IkappaB-, NF-kappaB-, and IL-8 pathways. We conclude that capsaicin can be proposed as a potential anti-inflammatory drug by inhibition of the production of IL-8 in H. pylori-infected gastric epithelium.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Capsaicin; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Signal Transduction

Topics: Adult; Aged; Bacterial Proteins; Cell Line; Coculture Techniques; Female; Gastric Mucosa; Genomic Islands; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lymphoma, B-Cell, Marginal Zone; Male; Middle Aged; Virulence Factors

Selective inhibition of proinflammatory chemokines such as IL-8 is an important approach to combat inflammatory and infection diseases. Previous studies suggested that interaction of transcription factors NFkappaB repressing factor (NRF) and NFkappaB play a crucial role in activation of IL-8 gene expression. In a search for a specific inhibitor of IL-8 expression, we applied tandem affinity purification to investigate interaction of NRF and NFkappaB p65 in cells. We identified a synthetic peptide corresponding to aa 223-238 of NRF interfering with binding of endogenous p65 to NRF. Furthermore, nucleofection experiments were established to introduce this inhibitory peptide into the nucleus of IL-1 stimulated human cervical and Helicobacter pylori infected gastric epithelial cells. Our data clearly show that the specific peptide disturbing NRF/NFkappaB interaction is able to significantly decrease endogenous IL-8 gene transcription in response to IL-1 or Helicobacter pylori infection. Thus, our study provides novel insights into NRF and NFkappaB interaction in vivo and may facilitate the design of new anti-IL-8 drugs based on novel strategies.

Topics: Cell Line, Tumor; DNA-Binding Proteins; Epithelial Cells; Gene Expression Profiling; HeLa Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; NF-kappa B; Peptides; Protein Binding; Repressor Proteins; Reverse Transcriptase Polymerase Chain Reaction; Transcription, Genetic

Recently, it has been shown that serine proteases derived from microorganisms stimulate epithelial cells to produce inflammatory mediators through protease-activated receptor (PAR). We investigated the involvement of PAR2 in the interleukin (IL)-8 production by Helicobacter pylori-infected gastric epithelial cells.. Human gastric epithelial cells, MKN45 cells, were used. The expression of PAR2 was assessed by real-time PCR and immunocytochemistry, and IL-8 protein was measured by an enzyme-linked immunosorbent assay. PAR2 mRNA and protein were constitutively expressed on unstimulated MKN45 cells. The treatment of cells with H. pylori resulted in a significant increase in PAR2 expression. In addition, trypsin (a natural PAR2 agonist), SLIGKV amide (a synthetic PAR2 agonist), H. pylori live bacteria or H. pylori culture supernatant significantly induced IL-8 production from MKN45 cells. H. pylori-induced IL-8 production was inhibited by nafamostat mesilate (a serine protease inhibitor), neutralizing antibody to PAR2 and in PAR2-deficient cells treated with siRNA.. These results reveal that H. pylori-derived protease activates gastric epithelial cells to produce inflammatory cytokines through PAR2, suggesting an important role for PAR2 in the modulation of gastric inflammation associated with H. pylori.

Topics: Antibodies; Benzamidines; Cell Line, Tumor; Epithelial Cells; Gastric Mucosa; Gene Expression; Guanidines; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Receptor, PAR-2; RNA, Small Interfering; Serine Proteinase Inhibitors; Transfection

Helicobacter species has been shown to be commonly present in extragastric human organs by polymerase chain reaction (PCR). To date, a few studies have reported that infection by Helicobacter pylori (H. pylori) was a risk factor for pancreatic malignancies, but this was not investigated very well. Therefore, we examined effects of H. pylori infection on human pancreatic cancer cells.. Interleukin (IL)-8 and vascular endothelial growth factor (VEGF) secretions by human pancreatic cancer cells which were co-cultured with H. pylori, were measured by enzyme-linked immunosorbent assay (ELISA). We then examined whether activities of proliferation factors nuclear factor-kappaB (NF-kappaB), activator protein-1 (AP-1), and serum response element (SRE) of human pancreatic cancer cells were increased by H. pylori infection. Furthermore, we examined cytotoxin-associated gene A protein (CagA) secretion into pancreatic cancer cells using Western blotting.. IL-8 and VEGF secretion levels and activities of proliferation factors NF-kappaB, AP-1, and SRE of human pancreatic cells increased by H. pylori infection. Moreover, CagA secretion into pancreatic cancer cells was confirmed by Western blotting.. Helicobacter pylori infection of human pancreatic cells may increase malignant potential of pancreatic cells, which seems to involve the same mechanisms as in gastric cancer cells.

Topics: Antigens, Bacterial; Bacterial Proteins; Blotting, Western; Cell Line, Tumor; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; Pancreatic Neoplasms; Serum Response Element; Transcription Factor AP-1; Vascular Endothelial Growth Factor A

Helicobacter pylori infection is associated with increased gastric epithelial cell turnover and non-cardia gastric cancer. Cell cycle progression is dependent on the proteasomal degradation of p27, a cyclin-dependent kinase inhibitor and gastric tumor suppressor, following ubiquitination mediated by Skp2. c-Myc is a transcriptional repressor of p27 and also a target of Skp2. In vitro, H. pylori decreases p27 protein post-translationally. We aimed to determine how p27 is regulated by H. pylori in vivo. The effect of eradicating H. pylori on gastric epithelial p27, Skp2, and c-Myc proteins and mRNA was investigated in 22 patients with chronic gastritis, by immunohistochemistry and laser capture microdissection. The percentage of gastric antral epithelial cells expressing p27 protein was significantly higher after eradication of H. pylori (mean+/-s.e.m. 37+/-2.4% pre-eradication vs 55+/-2.8% post-eradication; P<0.001), while Skp2 and c-Myc protein-expressing cells were lower (Skp2: 35+/-3.8 vs 23+/-2.6%, P=0.009; c-Myc: 47+/-3.6 vs 30+/-3.8%, P<0.001). mRNA expressions of p27, Skp2, and c-Myc (normalized for 18SrRNA) were not changed by H. pylori eradication. H. pylori increases c-Myc and decreases gastric epithelial p27 protein expression in association with increased expression of Skp2, the regulator of p27's ubiquitin ligase complex. H. pylori may influence cell cycle progression and carcinogenesis through post-translational effects on specific gene expression.

Topics: 2-Pyridinylmethylsulfinylbenzimidazoles; Adult; Aged; Amoxicillin; Anti-Bacterial Agents; Anti-Ulcer Agents; Chronic Disease; Clarithromycin; Cyclin-Dependent Kinase Inhibitor p27; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Lansoprazole; Male; Middle Aged; Omeprazole; Proteins; Proto-Oncogene Proteins c-myc; Proton Pump Inhibitors; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; S-Phase Kinase-Associated Proteins

The unavailability of human gastric cell lines representative of the normal gastric epithelial function such as polarized monolayer restricts the application of cell culture system in approaching the field of Helicobacter pylori (H. pylori) infected gastric mucosa models. The present investigation aimed at assessing the usefulness of NCI-N87 cell line as an adequate cellular model to study the pathophysiology of human H. pylori infection.. For the identification of epithelial phenotypes at low magnification, cells were observed on a phase-contrast microscope and confocal microscope. Transepithelial resistance (TER) was measured on NCI-N87 cells seeded on Transwell to identify monolayer polarity two or three times a week after confluency. The IL-8 level was determined by ELISA at 24 hours after the administration of HP60190 and IL-1 alpha on NCI-N87 cells. IL-8 level was compared in both upper and lower well with the control.. A monolayer phenotype was observed in NCI-N87 cell lines by using confocal microscope. TER was measured as 400-500 (Omega x cm2) at two or three weeks after cell culture. In NCI-N87 cell lines, IL-8 level was significantly increased after 24 hour compared to control, and was prominent in the lower well.. These results suggest that NCI-N87 cell line may be useful in H. pylori infected gastric mucosa model.

Topics: Cell Line; Enzyme-Linked Immunosorbent Assay; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Microscopy, Confocal; Microscopy, Phase-Contrast; Phenotype

To study the expression and significance of interleukin-8 (IL-8), cyclooxygenase-2 (COX-2) and trefoil family factor 1 (TFF1) in the remnant stomach mucosa.. Patients after gastrectomy were examined by upper gastrointestinal endoscopy. Biopsy specimens were obtained from stoma and the greater curvature of the upper corpus to be assessed for Hp (by H.E. and Giemsa staining) and conduct real-time semi-quantitative PCR. mRNA was extracted from the biopsy specimens to determine the IL-8, COX-2 and TFF1 gene mRNA levels by real-time PCR method.. In the stoma, COX-2 level in Hp-positive patients was significantly higher than that in Hp-negative patients, but the difference of IL-8 levels between them was not significant. In the corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients. In Hp-negative patients, IL-8 and COX-2 levels in the stoma were significantly higher in B II anastomosis than in B I anastomosis cases; COX-2 level in the stoma was significantly higher in B II anastomosis than in B I anastomosis cases, but the difference of IL-8 levels between them was not significant. TFF1 level in the remnant stomach mucosa showed no significant difference between Hp-positive and Hp-negative patients.. Hp infection and bile reflux are important risk factors for the secondary stomach carcinogenesis. Expression of IL-8 and COX-2 in the remnant stomach mucosa is related to the risk of secondary stomach carcinogenesis. The relationship between the TFF1 expression and secondary stomach carcinogenesis in the remnant stomach mucosa is still unclear and should further be studied.

Topics: Adult; Aged; Aged, 80 and over; Cyclooxygenase 2; Female; Gastrectomy; Gastric Mucosa; Gastric Stump; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Risk Factors; RNA, Messenger; Stomach Neoplasms; Trefoil Factor-1; Tumor Suppressor Proteins

Recent reports have shown an upregulation of macrophage migration inhibitory factor (MIF) during gastric ulcer development in a rat model and elevated counts of MIF-positive cells in biopsies from Helicobacter pylori-infected patients. H. pylori infection is a proven cofactor in humans causing gastritis and gastric ulcers. The aim of this study was to characterize MIF expression in human gastric epithelial cells in response to H. pylori.. MIF mRNA and MIF protein expression was detected in human gastric epithelial cell lines after stimulation with proinflammatory cytokines or infection with H. pylori (cagA+/vacA+) using real-time reverse transcriptase-polymerase and enzyme-linked immunosorbent assay. Interleukin-8 secretion was measured as positive control. MIF mRNA and MIF protein expression was assessed in H. pylori-positive and -negative human gastric biopsy samples.. While interleukin-8 mRNA expression and interleukin-8 secretion were upregulated in gastric epithelial cells in vitro after H. pylori infection, no changes in MIF mRNA expression and MIF secretion could be detected. We found no significant differences in MIF expression in total RNA extracted from gastric biopsy tissue when comparing H. pylori-positive to control patients. Likewise, MIF protein expression in gastric epithelium was unaffected by H. pylori infection as compared to uninfected tissue.. While an increased MIF expression and positive effects of MIF blockade in ulcer healing have been shown in a rodent model and elevated numbers of MIF-positive cells have been found in H. pylori-infected human tissue, we herein could not confirm any differences in human gastric epithelial MIF expression and secretion after H. pylori infection in vitro and in vivo.

Topics: Biopsy; Cell Line; Enzyme-Linked Immunosorbent Assay; Epithelium; Gastric Mucosa; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Macrophage Migration-Inhibitory Factors; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Accumulating evidence indicates that interleukin-8 (IL-8) plays a major role in the mucosal inflammation caused by Helicobacter pylori infection. The purpose of the present study was to examine whether Lactobacillus gasseri OLL2716 (LG21) can inhibit the H. pylori-induced production of IL-8.. A coculture system including MKN45 cells, H. pylori, and LG21 was established for an in vitro analysis. Biopsy specimens were obtained from H. pylori-infected human subjects consisting of 19 men and six women.. When LG21 was 1/100 less than H. pylori in a coculture system, LG21 significantly suppressed both the IL-8 mRNA and protein generation in the coculture. Live, but not heat- or UV-treated LG21, could exert the suppressive effect. However, this amount of LG21 could not suppress either the adhesion of H. pylori to the cell surface or the IL-8 production by tumor necrosis factor-alpha, which induces IL-8 generation through the activation of the transcription. These results thus suggest that LG21 suppresses an event leading to IL-8 production, which is specific for H. pylori-induced IL-8 generation, and this event is located upstream from the IL-8 transcription but downstream from the adhesion. The measurement of the IL-8 level using gastric biopsy specimens from H. pylori-infected subjects demonstrated that LG21 also suppresses the production of IL-8 in the gastric mucosa.. Live LG21 were found to suppress H. pylori-induced IL-8 production in both a gastric cell line and within gastric mucosa.

Topics: Adult; Aged; Animals; Biopsy; Cell Adhesion; Cell Line; Coculture Techniques; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactobacillus; Male; Mice; Middle Aged; NF-kappa B; Signal Transduction

To find out if a functional promoter polymorphism in the IL-8 gene along with cagA status and polymorphisms in vacA gene influence the type of diseases in Iranian patients infected by H pylori.. IL-8 -251 A/T polymorphism was genotyped by oligonucleotide allele specific PCR (ASO-PCR) in a sample of 233 patients with H pylori infection undergoing upper gastrointestinal endoscopy. The presence of cagA gene and polymorphisms in vacA gene was also determined by PCR. Association of these genetic polymorphisms with the development of gastritis, peptic ulcers as well as gastric cancer was tested.. When the patients with different clinical manifestations were compared according to the presence of cagA gene or various vacA genotypes, only the vacA genotypes were significantly different among gastritis, peptic ulcer and gastric cancer patients (chi 2 = 17.8; P = 0.001). Furthermore, there was a significant difference in the frequency of IL-8 -251 A/T genotypes between patients with gastric cancer and benign diseases (chi 2 = 10.47; P = 0.005).. The IL-8 -251 A/T polymorphism and the polymorphisms in H pylori vacA gene are involved in limiting the infection outcome to gastritis and peptic ulcer or in favoring cancer onset in Iranian patients.

Topics: Adult; Alleles; Antigens, Bacterial; Bacterial Proteins; Female; Gastrointestinal Diseases; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Iran; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Genetic; Treatment Outcome

With the steadily increasing occurrence of antibiotic resistance in bacteria, there is a great need for new antibacterial compounds. The approach described here involves targeting virulence-related bacterial type IV secretion systems (TFSSs) with small-molecule inhibitors. The cag TFSS of Helicobacter pylori was chosen as a model, and novel inhibitors directed against the cag VirB11-type ATPase Cagalpha were identified. The cag genes encode proteins that are components of a contact-dependent secretion system used by the bacterium to translocate the effector molecule CagA into host cells. Translocated CagA is associated with severe gastritis, and carcinoma. Furthermore, functional TFSSs and immunodominant CagA play a role in interleukin (IL)-8 induction, which is an important factor for chronic inflammation. Inhibitors of Cagalpha were identified by high-throughput screening of chemical libraries that comprised 524 400 small molecules. The ATPase activity of Cagalpha was inhibited by the selected compounds in an in vitro enzymic assay using the purified enzyme. The most active compound, CHIR-1, reduced TFSS function to an extent that cellular effects on AGS cells mediated by CagA were virtually undetectable, while reduced levels of IL-8 induction were observed. Gastric colonization by CHIR-1-pre-treated bacteria was found to be impaired in a dose-dependent manner using a mouse model of infection. Small-molecule Cagalpha inhibitors, the first described inhibitors of a TFSS, are potential candidates for the development of new antibacterial compounds that may lead to alternative medical treatments. The compounds are expected to impose weak selective pressure, since they target virulence functions. Moreover, the targeted virulence protein is conserved in a variety of bacterial pathogens. Additionally, TFSS inhibitors are potent tools to study the biology of TFSSs.

Topics: Adenosine Triphosphatases; Animals; Anti-Bacterial Agents; Antigens, Bacterial; Bacterial Proteins; Cell Line, Tumor; Colony Count, Microbial; Drug Evaluation, Preclinical; Enzyme Inhibitors; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mice; Microscopy, Confocal; Microscopy, Fluorescence; Virulence

Expression of the Helicobacter pylori outer membrane protein HopH is regulated by phase variation within a CT dinucleotide repeat motif of the hopH gene.. To investigate the importance of HopH for bacterial pathogenicity, we performed a detailed functional genomic and population-based genetic characterization of this contingency locus.. Sequencing of hopH in H. pylori strains from 58 patients revealed that the hopH "on" genotype is linked to bacterial virulence determinants, such as the vacAs1, vacAm1, babA2, and, most strongly, cagA genotypes. hopH mutagenesis resulted in reduced bacterial adherence to gastric epithelia in vitro. Complementation of hopH in trans restored the adherence properties of hopH mutants. Although HopH has been previously linked to proinflammatory epithelial signaling, hopH mutagenesis did not alter epithelial interleukin-8 secretion in vitro. Comparative epithelial gene-expression profiling by cDNA microarrays revealed no significant differences between the wild-type-specific and hopH mutant-specific transcriptomes. By contrast, a large set of genes was differentially regulated in a cag pathogenicity island-dependent manner.. An in-frame hopH gene may be linked to gastroduodenal diseases because of its association with other virulence factors or increased bacterial adherence and colonization. The strong linkage with cagA indicates that HopH may contribute to the fitness of cagA-positive strains in vivo.

Topics: Adhesins, Bacterial; Amino Acid Sequence; Antigens, Bacterial; Bacterial Adhesion; Bacterial Outer Membrane Proteins; Bacterial Proteins; Base Sequence; Cell Line, Tumor; DNA, Bacterial; Gastric Mucosa; Gene Expression Profiling; Genetic Complementation Test; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Molecular Sequence Data; Mutagenesis; Oligonucleotide Array Sequence Analysis; Polymorphism, Genetic; RNA, Messenger; Sequence Alignment; Virulence

Helicobacter pylori infection causes inflammation and increases the expression of IL-8 in human gastric epithelial cells. H. pylori activates NF-kappaB and AP-1, essential transcriptional factors in H. pylori-induced IL-8 gene transcription. Although colonization creates a local oxidative stress, the molecular basis for the transition from infection to the expression of redox-sensitive cytokine genes is unknown. We recently reported that the expression of apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE-1/Ref-1), which repairs oxidative DNA damage and reductively activates transcription factors including AP-1 and NF-kappaB, is increased in human gastric epithelia during H. pylori infection. In this study, we examine whether APE-1/Ref-1 functions in the modulation of IL-8 gene expression in H. pylori-infected human gastric epithelial cells. Small interfering RNA-mediated silencing of APE-1/Ref-1 inhibited basal and H. pylori-induced AP-1 and NF-kappaB DNA-binding activity without affecting the nuclear translocation of these transcription factors and also reduced H. pylori-induced IL-8 mRNA and protein. In contrast, overexpression of APE-1/Ref-1 enhanced basal and H. pylori-induced IL-8 gene transcription, and the relative involvement of AP-1 in inducible IL-8 promoter activity was greater in APE-1/Ref-1 overexpressing cells than in cells with basal levels of APE-1/Ref-1. APE-1/Ref-1 inhibition also reduced other H. pylori-induced chemokine expression. By implicating APE-1/Ref-1 as an important regulator of gastric epithelial responses to H. pylori infection, these data elucidate a novel mechanism controlling transcription and gene expression in bacterial pathogenesis.

Topics: Cell Line; DNA-(Apurinic or Apyrimidinic Site) Lyase; Electrophoretic Mobility Shift Assay; Gastric Mucosa; Gene Expression; Gene Silencing; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lasers; Microdissection; NF-kappa B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; Transcription Factor AP-1; Transcription, Genetic

Ghrelin and leptin are endogenous peptides that have been implicated in the control of food intake, energy homeostasis and body weight gain. Although the stomach is the major source of circulating ghrelin and partly contributes also to plasma leptin, controversy exists over the influence of gastric Helicobacter pylori (Hp) infection on the ghrelin and leptin release. To resolve this controversy, plasma immunoreactive ghrelin and leptin levels were determined in Hp-positive and Hp negative children (N=60) and in adults (N=120) and daily concentrations of these hormones were measured at 2 h intervals before and after meals. Serum levels of ghrelin and leptin as well as gastrin were measured by RIA. Hp status was assessed using (13)C-urea breath test (UBT) and serology. Children with negative UBT showed significantly higher basal serum levels of ghrelin and lower concentrations of leptin than those with positive UBT. Adults without Hp infection also showed significantly higher fasting serum levels of ghrelin and lower levels of leptin than those in Hp infected subjects. In adults, especially without Hp infection, plasma levels of ghrelin showed a marked rise before the meal and sudden decrease following the food intake, while plasma leptin did not showed significant meal-related alterations, but in general its level was significantly higher in Hp positive than Hp negative subjects. Serum gastrin concentrations were significantly elevated in both Hp positive children and adults and these levels were significantly lower in Hp negative subjects. We conclude that Hp infection in children and adults causes a marked reduction in plasma levels of ghrelin, while increasing plasma levels of leptin and gastrin. These alterations in plasma levels of gastric originated appetite-controlling hormones in Hp infected children and adults may contribute to the alterations of the appetite and dyspeptic symptoms observed in these subjects.

Topics: Adolescent; Adult; Child; Eating; Female; Gastrins; Ghrelin; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Leptin; Male; Middle Aged; Tumor Necrosis Factor-alpha

To determine the concentrations of leptin in plasma and gastric fundic mucosa in humans, with reference to Helicobacter pylori (H pylori) infection, and their association with gastric mucosal levels of interleukin (IL)-1beta, IL-6 and IL-8.. Plasma leptin concentrations were determined in 135 outpatients with non-ulcer dyspepsia, consisting of 95 H pylori-infected and 40 uninfected subjects, and 13 patients before and after cure of the infection with anti-H pylori regimen. Using biopsy samples that were endoscopically obtained from the middle corpus along the greater curvature, gastric leptin contents were measured by radioimmunoassay and the mucosal concentrations of IL-1beta, IL-6 and IL-8 were measured by enzyme linked immunosorbent assay. We also analysed the expression of leptin in the fundic mucosa by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry.. The mucosal levels of leptin in the fundic mucosa of H pylori-infected patients were significantly higher than those of uninfected patients. The amount of gastric leptin correlated positively with the mucosal levels of IL-1beta and IL-6, but not IL-8. Circulating leptin correlated with body mass index, but not with H pylori status, and there was no change in plasma leptin levels following cure of the infection. Leptin immunoreactive cells were noted in the lower half of the fundic glands, and its expression of messenger ribonucleic acid in the oxyntic mucosa was detected by RT-PCR.. Leptin production is enhanced in H pylori-infected gastric mucosa. Gastric leptin may be involved in immune and inflammatory response during H pylori infection, through interaction with proinflammatory cytokines.

Topics: Adult; Aged; Aged, 80 and over; Female; Gastric Fundus; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Leptin; Male; Middle Aged; RNA, Messenger

Helicobacter pylori infection is associated with variable clinical outcomes, including gastroduodenal diseases, and genetic factors may be relevant in this process.. We investigated the effects of an interleukin 8 (IL-8) gene polymorphism on the risk of gastroduodenal diseases, the degree of H pylori induced gastritis, and IL-8 gene transcription.. The study was performed in 244 healthy control subjects and 690 H pylori positive patients with non-cardia gastric cancer, gastric ulcer, duodenal ulcer, or gastritis.. We identified the IL-8 -251 A/T polymorphism by direct sequence analysis, and measured the gastritis score and serum pepsinogen (PG). The transcriptional promoter activity of the IL-8 gene was assessed by luciferase assay.. IL-8 -251A was associated with a higher risk of gastric cancer and gastric ulcer. Patients carrying IL-8 -251A showed an increased risk of gastric cancer (odds ratios (OR) 2.01 (95% confidence interval (CI) 1.38-2.92)) and gastric ulcer (OR 2.07 (95% CI 1.37-3.12)). Compared with patients younger than 49 years, atrophy and metaplasia scores in the antrum were significantly higher and the PG I/II ratio significantly lower in -251A carriers than in T/T carriers. In the in vitro assay, IL-8 -251A showed enhanced promoter activity in response to IL-1beta or tumour necrosis factor alpha.. The IL-8 -251A allele may be associated with progression of gastric atrophy in patients with H pylori infection, and may increase the risk of gastric cancer and gastric ulcer in Japanese people.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Asian People; Duodenal Ulcer; Female; Gastritis; Genetic Predisposition to Disease; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Polymorphism, Genetic; Promoter Regions, Genetic; Stomach Diseases; Stomach Neoplasms; Stomach Ulcer

Helicobacter pylori infection is one of the most common gastrointestinal infections worldwide. Although the majority of the infected individuals remain asymptomatic carriers of the bacteria, approximately 15% develop peptic ulcers, which are most prevalent in the duodenum. H. pylori induce a vigorous immune response which, however, fails to clear the infection. Instead, the chronic inflammation that arises in the infected gastroduodenal mucosa may be involved in the development of H. pylori-associated peptic ulcers. We have previously shown that duodenal ulcer (DU) patients have a significantly lower epithelial cytokine, e.g. IL-8, response in the duodenum than asymptomatic (AS) carriers. In this study we have further investigated the mechanisms behind this finding, i.e. whether it can be explained by bacterial factors, down-regulation of epithelial cytokine production by regulatory T cells, or an impaired ability of the duodenal epithelium in DU patients to produce cytokines. Gastric AGS, and intestinal T84 epithelial cell lines were stimulated with H. pylori strains isolated from DU patients and AS carriers, respectively. All strains were found to induce comparable cytokine and cytokine receptor expression in epithelial cells. Regulatory T cells (CD4+ CD25(high)), isolated from human peripheral blood and cocultured with H. pylori stimulated AGS cells, were found to slightly suppress H. pylori-induced epithelial cytokine production. Furthermore, primary cultures of duodenal epithelial cells from DU patients were found to produce markedly lower amounts of cytokines than epithelial cells isolated from AS carriers. These results suggest that the lower epithelial cytokine responses in the duodenum of DU patients, which may be of importance for the pathogenesis of H. pylori-induced duodenal ulcers, most likely can be explained by host factors, i.e. mainly a decreased ability of the duodenal epithelium to produce cytokines, but possibly partly also down-regulation by regulatory T cells.

Topics: CD4-Positive T-Lymphocytes; Cell Line; Coculture Techniques; Down-Regulation; Duodenal Ulcer; Epithelial Cells; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Receptors, Cytokine; Receptors, Interleukin-2; T-Lymphocytes; Transforming Growth Factor beta

The roles of the virD4 and the cagG genes in the cag pathogenicity island of Helicobacter pylori for gastroduodenal pathogenesis are unclear and their roles in vivo have not been examined.. Seven week old male Mongolian gerbils were inoculated with the wild type H pylori TN2GF4, its isogenic virD4, or cagG mutants. Animals were sacrificed at 4, 12, and 24 weeks after inoculation. Gastric inflammation and H pylori density were evaluated by histology, inflammatory response (as measured by interleukin (IL)-1beta mRNA levels), proliferative activity (as assessed by 5'-bromo-2'deoxyuridine labelling indices), and host systemic reaction (as measured by anti-H pylori IgG antibody).. Degree of gastric inflammation, proliferative activity, and mucosal IL-1beta mRNA levels remained low throughout the first 12 weeks in gerbils infected with the virD4 mutants. Degree of gastric inflammation and proliferative activity increased at 24 weeks with the virD4 mutants reaching levels comparative with those seen at four weeks with the wild-type strains. Mucosal IL-1beta mRNA levels were also increased at 24 weeks with the virD4 mutants and levels at 24 weeks were similar between the wild-type and virD4 mutants. In contrast, gerbils infected with the cagG mutants had reduced ability to colonise gerbils, and no or little gastric inflammation or proliferative activity was observed.. Loss of the virD4 gene temporally retarded but did not abrogate gastric inflammation. Loss of the cagG gene abolished gastric inflammation partially via reduced ability to colonise gerbils. Unknown factors related to the type IV secretion system other than CagA may influence gastric inflammation.

Topics: Animals; Bacterial Proteins; Cell Division; Cells, Cultured; Disease Models, Animal; Gastric Mucosa; Gastritis; Genes, Bacterial; Genomic Islands; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Interleukin-1; Interleukin-8; Male; RNA, Messenger; Virulence; Virulence Factors

To study the expressions and the significance of interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the remnant stomach.. Fifty-eight patients with gastrectomy were examined by upper gastrointestinal endoscopy. Two biopsy specimens were obtained from the stoma and the upper corpus gastric mucosa in the remnant stomach. mRNA was extracted from biopsy specimens to measure the IL-8 and COX-2 gene mRNA levels by real-time PCR method.. IL-8 and COX-2 levels were higher in stoma than in corpus, IL-8 levels in BI anastomosis were significantly higher in stoma than in corpus (P< 0.05). In Hp-negative patients, IL-8 and COX-2 levels in stoma were significantly higher in BII anastomosis than in BI anastomosis (P < 0.05). In Hp-positive patients, IL-8 and COX-2 levels in stoma showed no significant differences between BII anastomosis and BI anastomosis. In corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients, (P < 0.05), including in BI anastomosis and in BII anastomosis.. The risk of the secondary stomach carcinogenesis in stoma after distal gastrectomy is higher than that in corpus; The types of anastomosis may influence the risk for the secondary stomach carcinogenesis in the remnant stomach mucosa.

Topics: Adult; Aged; Aged, 80 and over; Female; Gastric Mucosa; Gastric Stump; Gastroenterostomy; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Prostaglandin-Endoperoxide Synthases; RNA, Messenger; Stomach Neoplasms

Macrolide antibiotics have an anti-inflammatory effect by suppressing lipopolysaccharide-induced IL-8 production. IL-8 secretion from monocytes is observed in Helicobacter pylori infection. Although cag gene products are known to induce IL-8 secretion, whether other bacterial substances can initiate the reaction is not determined. In this study, we show that clarithromycin induced down-regulation of Toll-like receptor 4 expression and did not lead to a decrease in IL-8 production and H. pylori lipopolysaccharide. However, Toll-like receptor 4 activation was possibly not the main cause in the induction of inflammation during H. pylori infection.

Topics: Anti-Bacterial Agents; Clarithromycin; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lipopolysaccharides; Macrolides; Membrane Glycoproteins; Monocytes; Receptors, Cell Surface; Toll-Like Receptor 4; Toll-Like Receptors

The aim of this analysis was to evaluate the effects of the presence of the IL-1RA gene polymorphism and H. pylori infection on markers of a systemic inflammatory response taking into account virulence markers of this infection. Serum concentrations of interleukin (IL)-6, IL-8, and tumour-necrosis factor (TNF)-alpha of 479 occasional blood donors were not statistically significantly higher in subjects having antibodies against H. pylori, or more specifically against CagA and VacA, and being homozygous for the pro-inflammatory IL-1RN*2 allele compared to others after adjustment for covariates. The findings suggest that the possible pro-inflammatory effect of the IL-1RN*2 allele in combination with H. pylori infection is limited to the mucosal level.

Topics: Adult; Aged; Female; Germany; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-6; Interleukin-8; Male; Middle Aged; Polymorphism, Genetic; Receptors, Interleukin-1; Sialoglycoproteins; Tumor Necrosis Factor-alpha

CD74, or the class II MHC-associated invariant chain, is best known for the regulation of Ag presentation. However, recent studies have suggested other important roles for this protein in inflammation and cancer studies. We have shown that CD74 is expressed on the surface of gastric cells, and Helicobacter pylori can use this receptor as a point of attachment to gastric epithelial cells, which lead to IL-8 production. This study investigates the ability of H. pylori to up-regulate one of its receptors in vivo and with a variety of gastric epithelial cell lines during infection with H. pylori. CD74 expression was increased dramatically on gastric biopsies from H. pylori-positive patients and gastric cell lines exposed to the bacteria. Gastric cells exposed to H. pylori-conditioned medium revealed that the host cell response was responsible for the up-regulation of CD74. IL-8 was found to up-regulate CD74 cell surface expression because blocking IL-8Rs or neutralizing IL-8 with Abs counteracted the increased expression of CD74 observed during infection with H. pylori. These studies demonstrate how H. pylori up-regulates one of its own receptors via an autocrine mechanism involving one of the products induced from host cells.

Topics: Antigens, Differentiation, B-Lymphocyte; Bacterial Adhesion; Cell Line; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Histocompatibility Antigens Class II; Humans; In Vitro Techniques; Interleukin-8; Neutralization Tests; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Up-Regulation

To explore the relationship between helicobacter pylori (Hp) infection and expression of interleukin-8 (IL-8) in the remnant stomach mucosa.. Fifty-eight patients with gastrectomy were examined by upper gastrointestinal endoscopy. Biopsy specimens were obtained from stoma and the greater curvature of the upper corpus to detect Hp infection (by H.E. and Giemsa staining) and IL-8 expression (by real-time semi-quantitative RT-PCR).. Hp infection was detected in 65.5% (38/58)of the remnant stomach mucosa. In 34 patients with Billroth I (BI) anastomosis, IL-8 in corpus was significantly higher in Hp-positive patients than that in Hp-negative patients (0.11+/-0.07 vs 0.02+/-0.01, P<0.05). In 24 patients with Billroth II (BII) anastomosis, IL-8 in stoma and corpus in Hp-positive patients was significantly higher than that in Hp-negative patients (P<0.05).. Hp infection induces IL-8 expression in remnant stomach mucosa. In corpus, IL-8 mRNA expression is mainly related with Hp infection, while in stoma, IL-8 mRNA expressions may be related with bile reflux besides Hp infection.

Topics: Adult; Aged; Aged, 80 and over; Endoscopes, Gastrointestinal; Female; Gastric Mucosa; Gastric Stump; Gastroenterostomy; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; RNA, Messenger

Helicobacter pylori has been known to provoke gastric inflammation, ulceration, and DNA damage, based on which WHO defined H. pylori as a class I carcinogen. Although ginseng, the root of Panax ginseng C.A. Meyer, has been reported to possess antiadhesion or antimicrobial activity against H. pylori, in this study, we examined the protective effect of red ginseng extracts (RGE) against H. pylori-induced cytotoxicity and DNA damage. RGE significantly attenuated both H. pylori-induced DNA damage assessed by comet assay and apoptosis measured by DNA fragmentation. Inactivation of ERK1/2 signaling and attenuation of caspase-3 activation and PARP cleavage were revealed with RGE against H. pylori infection. RGE decreased H. pylori-stimulated IL-8 gene expression, which resulted from the transcriptional regression of NF-kappaB. In conclusion, RGE showed significant gastroprotective effects against H. pylori-associated gastric mucosal cell damage, suggesting that red ginseng could be used as a medicinal phytonutrient against H. pylori infection.

Topics: Apoptosis; Cell Division; Cell Line, Tumor; Cell Survival; Cytoprotection; DNA Damage; Gastric Mucosa; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Panax; Plant Extracts

To determine the role of host immune responses in H. pylori infection, we examined the relationship between gastric mucosal IL-8 levels and histological gastritis in patients with H. pylori infection. Biopsy tissue obtained from 99 patients were homogenizedand mucosal IL-8 levels measured by ELISA. The gastric mucosal IL-8 levels in both the antrum and corpus were higher in patients with H. pylori than in H. pyloi negativepatients. IL-8 levels in the corpus but not the antrum correlated with the severity of the atrophy. The IL-1B polymorphism had no influence on the degree of IL-8 production. These findings indicate that IL-8 production is independent of IL-1B polymorphisms and IL-8 may play an important role in the development of atrophic gastritis.

Topics: Adult; Aged; Aged, 80 and over; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Neutrophil Infiltration; Polymorphism, Genetic

Persistent interleukin-8 (IL-8) production contributes to chronic inflammation of the stomach. The proinflammatory IL-1beta polymorphisms, which enhance the cytokine production, are associated with increased risk of gastric cancer. The -251A/T polymorphism of the IL-8 promoter is involved in several human diseases. Particularly, the -251A is associated with decreased risk of colorectal cancer. We aimed to determine whether the -251 allele resulting in high IL-8 expression was associated with increased risk of gastric carcinoma.. The -251A/T promoters were cloned and analyzed by luciferase assay. Binding of nuclear proteins to the -251A/T promoters was analyzed by electrophoretic mobility shift assay. The -251A/T promoters were differentiated by PCR-RFLP. Comparison of gastric cancer risk between the -251A/T promoters was done by a case-control study.. The -251T allele possessed transcriptional activity 2- to 5-fold stronger than the -251A counterpart. Electrophoretic mobility shift assay showed that the -251A promoter had strong ability to bind to an unknown protein or multiprotein complex. The -251T allele was associated with increased risk of noncardia (P(trend) = 0.012) and cardia (P(trend) = 0.029) carcinomas. Gastric carcinoma patients with the low-risk AA genotype had a tendency to sustain intestinal-type carcinomas (chi(2) = 6.816; P = 0.033); however, the high-risk -251T allele was associated with >2-fold increased risk of diffuse-type (AA versus AT + TT: odds ratio, 2.52; 95% confidence interval, 1.16-5.49; P = 0.017) and mixed-type (AA versus AT + TT: odds ratio, 2.22; 95% confidence interval, 1.12-4.40; P = 0.019) carcinomas.. The IL-8 -251T allele is significantly associated with increased risk of gastric carcinoma, particularly the diffuse and mixed types in Chinese population.

Topics: Adult; Aged; Alleles; Base Sequence; Case-Control Studies; Cell Line, Tumor; Chi-Square Distribution; China; Female; Gene Frequency; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Odds Ratio; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Risk Factors; Stomach Neoplasms; Tumor Cells, Cultured

Gastric epithelial cell lines have been utilized extensively as tools to define aspects of the interactions between Helicobacter pylori and host epithelial cells. Fetal calf serum (FCS) is employed as a growth stimulant, but it is unclear whether this agent may in itself alter host responses.. Two gastric epithelial cell lines were utilized to ascertain the effects of varying FCS concentration on cellular responses following H. pylori infection. Media containing 0%, 5% or 10% FCS was added to cell lines prior to infection with H. pylori of defined genotype. Cellular interleukin (IL)-8 production was measured as a marker of cellular response. Effects of altered FCS upon cell viability were also determined by trypan blue exclusion.. Interleukin-8 production by AGS cells following H. pylori infection was not altered by variation of media FCS concentration. However, KATO-III cells produced greater amounts of IL-8 when media was FCS-free than at 5% or 10% FCS. Although cellular viability was not altered in AGS cells exposed to varied concentrations of FCS, viability was decreased in serum-free KATO-III cells, but not when cells were kept at 5% FCS.. Serum-derived factors alter cellular responses to H. pylori infection in a cell-line-dependent manner and impaired cellular viability may relate to this effect. However, the mechanisms for these observations are unclear and further work is now required to determine the nature of these important interactions.

Topics: Animals; Cattle; Cell Line, Tumor; Cell Survival; Fetal Blood; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8

Chronic inflammation induced by Helicobacter pylori infection is closely associated with epithelial cell proliferation and apoptosis, which are related to cellular turnover in gastric mucosa. Reg protein is a regenerating gene product and a potent growth factor for gastric mucosal cells, however, little is known regarding its association with the pathogenesis of H. pylori infection. The aim of this study was to investigate Reg protein production and its regulation in H. pylori-associated gastritis.. Gastric fundic biopsy samples were taken from patients with and without H. pylori infection. In vivo expression of Reg protein was examined by Western blotting and immunohistochemistry methods. The effects of interleukin (IL)-8 on Reg protein expression and transcriptional activation of the Reg gene in ECC10 cells were investigated by Western blotting and luciferase assays, respectively.. Reg expression was found localized in the deeper part of gastric fundic glands and clearly shown in chromogranin A-positive cells in the gastric corpus. Semiquantitative immunohistochemistry and Western blotting results for Reg expression were significantly associated with polymorphonuclear neutrophil activity and chronic inflammation of gastric mucosa. IL-8 production in the gastric mucosa was significantly augmented by H. pylori infection, while IL-8 dose-dependently stimulated Reg protein production and Reg promoter activity in vitro in cultured ECC10 cells.. The present study showed for the first time that Reg protein may be a potent stimulator of gastric epithelial cells in H. pylori-infected human gastric mucosa stimulated by IL-8. Further, our findings provide evidence of a novel link between Reg protein and H. pylori infection, which may help explain the molecular mechanisms underlying H. pylori-associated diseases, including gastric cancer.

Topics: Adult; Aged; Calcium-Binding Proteins; Cell Culture Techniques; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lithostathine; Male; Middle Aged; Nerve Tissue Proteins; Neutrophil Infiltration; Neutrophils

Host genetic susceptibility may influence gastric carcinogenesis caused by Helicobacter pylori infection. We aimed to clarify the relationship of interleukin (IL)-8 polymorphism with the risk of atrophic gastritis and gastric cancer. We examined IL-8 -251 T > A, IL-1B -511 C > T, and IL-1RN intron 2 polymorphisms in 252 healthy controls, 215 individuals with atrophic gastritis, and 396 patients with gastric cancer. We also investigated the effect of the IL-8 polymorphism on IL-8 production and histologic degree of gastritis in noncancerous gastric mucosa. Although no correlation was found in the analysis of the IL-1B and IL-1RN polymorphisms, IL-8 -251 A/A genotype held a higher risk of atrophic gastritis [odds ratio (OR), 2.35; 95% confidence interval (CI), 1.12-4.94] and gastric cancer (OR, 2.22; 95% CI, 1.08-4.56) compared with the T/T genotype. We also found that the A/A genotype increased the risk of upper-third location (OR, 3.66; 95% CI, 1.46-9.17), diffuse (OR, 2.79; 95% CI, 1.21-6.39), poorly differentiated (OR, 2.70; 95% CI, 1.14-6.38), lymph node (OR, 2.50; 95% CI, 1.01-6.20), and liver metastasis (OR, 5.63; 95% CI, 1.06-30.04), and p53-mutated (OR, 1.91; 95% CI, 1.13-3.26) subtypes of gastric cancer. The A/A and A/T genotypes were significantly associated with higher levels of IL-8 protein compared with the T/T genotype. Neutrophil infiltration score was significantly higher in the A/A genotype than in the T/T genotype. In conclusion, we showed that the IL-8 -251 T > A polymorphism is associated with higher expression of IL-8 protein, more severe neutrophil infiltration, and increased risk of atrophic gastritis and gastric cancer.

Topics: Adult; Aged; Aged, 80 and over; Atrophy; Case-Control Studies; Female; Gastritis; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Humans; Interleukin-8; Japan; Male; Middle Aged; Odds Ratio; Polymorphism, Genetic; Promoter Regions, Genetic; Risk Factors; Stomach Diseases; Stomach Neoplasms; Up-Regulation

In clinical settings, Lactobacillus johnsonii La1 administration has been reported to have a favorable effect on Helicobacter pylori-associated gastritis, although the mechanism remains unclear. We administered, continuously through the water supply, live La1 to H. pylori-infected C57BL/6 mice and followed colonization, the development of H. pylori-associated gastritis in the lamina propria, and the levels of proinflammatory chemokines macrophage inflammatory protein 2 (MIP-2) and keratinocyte-derived cytokine (KC) in the serum and gastric tissue over a period of 3 months. We documented a significant attenuation in both lymphocytic (P=0.038) and neutrophilic (P=0.003) inflammatory infiltration in the lamina propria as well as in the circulating levels of anti-H. pylori immunoglobulin G antibodies (P=0.003), although we did not observe a suppressive effect of La1 on H. pylori colonizing numbers. Other lactobacilli, such as L. amylovorus DCE 471 and L. acidophilus IBB 801, did not attenuate H. pylori-associated gastritis to the same extent. MIP-2 serum levels were distinctly reduced during the early stages of H. pylori infection in the La1-treated animals, as were gastric mucosal levels of MIP-2 and KC. Finally, we also observed a significant reduction (P=0.046) in H. pylori-induced interleukin-8 secretion by human adenocarcinoma AGS cells in vitro in the presence of neutralized (pH 6.8) La1 spent culture supernatants, without concomitant loss of H. pylori viability. These observations suggest that during the early infection stages, administration of La1 can attenuate H. pylori-induced gastritis in vivo, possibly by reducing proinflammatory chemotactic signals responsible for the recruitment of lymphocytes and neutrophils in the lamina propria.

Topics: Animals; Antibodies, Bacterial; Colony Count, Microbial; Follow-Up Studies; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Immunoglobulin G; Interleukin-8; Lactobacillus; Lymphocytes; Mice; Mice, Inbred C57BL; Neutrophil Infiltration

The risk factors for secondary stomach carcinogenesis after distal gastrectomy have not been evaluated in detail.. Using gastrointestinal endoscopy, we examined 112 patients who had undergone gastrectomy. Biopsy specimens were taken from the stoma and the upper corpus mucosa in the remnant stomach to examine the associations among Helicobacter pylori (H.pylori) infection, bile reflux, and the expressions of interleukin-8 (IL-8), cyclo-oxygenase-2 (COX-2), and trefoil factor family 1 (TFF1) genes in the stomach mucosa.. The IL-8 levels in the corpus mucosa were significantly higher in the H.pylori-positive patients than in the H.pylori-negative patients (P = 0.015). The IL-8 levels were significantly higher in the stomal mucosa than in the corpus mucosa in the H.pylori-positive patients (P = 0.047). The COX-2 levels in the corpus mucosa tended to be higher in the H.pylori-positive patients, but these levels were not significantly different in the stoma mucosa. The COX-2 levels in the corpus were significantly higher after Billroth II (BII) anastomosis than after Billroth I (BI) anastomosis (P = 0.041). TFF1 expression in the stoma was higher in the H.pylori-positive patients than in the H.pylori-negative patients, but the difference was not significant.. Both H.pylori infection and bile reflux increased IL-8 levels after BI anastomosis. Furthermore, COX-2 levels were higher after BII than after BI anastomosis. These indicators will become useful not only as biomarkers to predict the degree of inflammation in the stomach mucosa, but also as surrogate biomarkers to predict the risk of secondary stomach carcinogenesis in the remnant stomach mucosa.

Topics: Adult; Aged; Aged, 80 and over; Bile Reflux; Biomarkers, Tumor; Cyclooxygenase 2; Female; Gastrectomy; Gastric Stump; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Predictive Value of Tests; Risk Factors; Stomach Neoplasms; Trefoil Factor-1; Tumor Suppressor Proteins

To determine the role of interleukin (IL)-17 in gastric ulcerogenesis.. Thirty-six gastric ulcer (GU) patients and 29 non-ulcer (NU) patients were enrolled in this study. Mucosal biopsy samples were obtained from the gastric antrum and GU site during endoscopy. Samples were used in in situ stimulation for 48 h in the presence of 10 microg/mL phytohemagglutinin-P (PHA), histological examination, and Helicobacter pylori (H pylori) culture. IL-17 and IL-8 protein levels in culture supernatants were assayed by ELISA. IL-17 mRNA expression was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). H pylori cagA and vacA status was assessed by reverse hybridization using a line probe assay (LiPA). IL-8 levels in culture supernatants were assayed after AGS cells were co-cultured with H pylori strain 26,695 or recombinant human (rh) IL-17.. All 36 GU patients and 15 of 29 NU patients were found to be H pylori-positive, while 14 NU patients were H pylori-negative. All 51 H pylori strains from both GU and NU patients were cagA- and vacAs1/m1-positive. Antral mucosal tissues from H pylori-positive patients contained significantly (H pylori-positive NU patients: median 467 pg/mg/protein, range 53-2,499; H pylori-negative NU patients: median 104 pg/mg/protein, range 16-312, P< 0.0005) higher levels of IL-17 than those from uninfected patients. IL-17 levels at the ulcer site were significantly (ulcer site: median 1,356 pg/mg/protein, range 121-1,3730; antrum: median 761 pg/mg/protein, range 24-7,620, P< 0.005) higher than those at distant sites in the antrum. Biopsies from H pylori-positive GU and NU patients showed IL-17 mRNA expression in all samples whereas those from the antrum of the H pylori-negative controls showed no detectable expression. A significant correlation was seen between IL-17 and IL-8 levels at each biopsy site (ulcer: r = 0.62, P< 0.0001; antrum: r = 0.61, P< 0.0001) in GU patients. RhIL-17 and H pylori strain 26,695 each stimulated IL-8 production from AGS cells.. IL-17 may play an important role in the inflammatory response to H pylori colonization, and may ultimately influence the outcome of H pylori-associated diseases that arise within the context of gastritis.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Biopsy; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-8; RNA, Messenger; Stomach Ulcer

Oxidant-sensitive transcription factors, nuclear factor-kappaB (NF-kappaB), and activator protein-1 (AP-1) have been considered as the regulators of inducible genes such as chemokines. Since oxygen radicals are considered as an important regulator in the pathogenesis of Helicobacter pylori (H. pylori)-induced gastric ulceration and carcinogenesis, chemokines such as interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) may be regulated by NF-kappaB and/or AP-1. Ras, the upstream activator for mitogen-activated protein kinase (MAPK) and MAPK cascade regulate AP-1 activation. The present study aims to investigate whether H. pylori in a Korean isolate (HP99) induces the expression of chemokines (IL-8, MCP-1), which is regulated by Ras, MAPK, AP-1, and NF-kappaB in gastric epithelial AGS cells, and whether these transcriptional regulations of chemokines are inhibited by transfection with mutant genes for Ras (ras N-17), c-Jun (TAM-67), and IkappaBalpha (MAD-3) or treatment with MAPK inhibitors (U0126 for extracellular signal-regulated kinase or SB203580 for p38 kinase). In addition, virulence factors of HP99 were characterized by PCR analysis for the isolated DNA. As a result, HP99 is identified as cagA+, vacA s1b, m2, iceA1 H. pylori strain. HP99 induced a time-dependent expression of mRNA and protein for IL-8 and MCP-1 via mediation of MAPK, AP-1, and NF-kappaB. Transfection with mutant genes for Ras, c-Jun, and IkappaBalpha and treatment with MAPK inhibitors suppressed H. pylori-induced activation of transcription factors (NF-kappaB, AP-1) and expression of chemokines (IL-8, MCP-1) in AGS cells. In conclusion, Ras and MAPK cascade may act as the upstream signaling for the activation of AP-1 and NF-kappaB, which induce chemokine expression in H. pylori-infected AGS cells. Specific targeting of the activation of NF-kappaB and AP-1 may be effective for the prevention or treatment of gastric inflammation associated with H. pylori infection.

Topics: Cell Line, Tumor; Chemokine CCL2; DNA, Bacterial; Gastric Mucosa; Gene Expression Regulation, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mitogen-Activated Protein Kinases; NF-kappa B; Polymerase Chain Reaction; RNA, Messenger; Transcription Factor AP-1; Virulence Factors

Infection of epithelial cells by the microbial pathogen Helicobacter pylori leads to activation of the transcription factor nuclear factor kappaB (NF-kappaB), the induction of pro-inflammatory cytokine/chemokine genes, and the motogenic response (cell scattering). Here we report that H. pylori-induced NF-kappaB activation and the subsequent release of interleukin 8 (IL-8) are inhibited by curcumin (diferuloylmethane), a yellow pigment in turmeric (Curcuma longa L.). Our results demonstrate that curcumin inhibits IkappaBalpha degradation, the activity of IkappaB kinases alpha and beta (IKKalpha and beta), and NF-kappaB DNA-binding. The mitogen-activated protein kinases (MAPK), extracellular signal-regulated kinases 1/2 (ERK1/2) and p38, which are also activated by H. pylori infection, were not inhibited by curcumin. Further, the H. pylori-induced motogenic response was blocked by curcumin. We conclude that curcumin, due to inhibition of NF-kappaB activation and cell scattering, should be considered as a potential therapeutic agent effective against pathogenic processes initiated by H. pylori infection.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Line; Cell Movement; Curcumin; Dose-Response Relationship, Drug; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B

Gastric mucosal interleukin (IL)-8 levels are related to the presence of both the cag pathogenicity island (PAI) and OipA. We investigated the regions of the IL-8 promoter and the upstream signaling involved in IL-8 gene transcription.. We cocultured parental Helicobacter pylori and isogenic oipA, hopZ, or cagE gene knockout mutants with gastric cancer cells. The regulatory sites in the IL-8 promoter were examined by luciferase reporter gene assay, electrophoretic mobility shift assays, and immunoblot analyses. Phosphorylated signal transducers and activators of transcription 1 (STAT1) levels in the antral gastric mucosa were measured by enzyme-linked immunosorbent assay.. Maximal H. pylori -induced IL-8 gene transcription required the presence of the interferon-stimulated responsive element (ISRE)-like element, nuclear factor (NF)-kappa B and activator protein (AP)-1 binding sites. In vitro studies showed that OipA and the cag PAI were involved in inducing interferon regulatory factor (IRF)-1 to bind and activate the ISRE-like element and that the cag PAI, but not OipA, was involved in activating AP-1 and NF-kappa B. Both in vitro and in vivo studies showed that OipA, but not the cag PAI, was involved in STAT1 phosphorylation, as upstream signaling of IRF-1.. OipA and the cag PAI are both necessary for full activation of the IL-8 promoter but act via different pathways that diverge upstream of IRF-1 where only OipA is involved in the STAT1-IRF1-ISRE pathway. The mucosal inflammatory response to H. pylori infection is complex and involves different pathways converging on the IL-8 promoter.

Topics: Antigens, Bacterial; Bacterial Adhesion; Bacterial Outer Membrane Proteins; Bacterial Proteins; Cell Line, Tumor; DNA-Binding Proteins; Epithelial Cells; Gastritis; Gene Expression Regulation; Genes, Reporter; Helicobacter Infections; Helicobacter pylori; Humans; Interferon Regulatory Factor-1; Interleukin-8; Mutagenesis; NF-kappa B; Phosphoproteins; Promoter Regions, Genetic; Signal Transduction; STAT1 Transcription Factor; Stomach Neoplasms; Trans-Activators; Transcription Factor AP-1; Transcriptional Activation

Low recurrence of gastritis is seen in patients infected with Helicobacter pylori carrying the type II urease B gene, compared with H. pylori carrying types I and III. The underlying mechanism has been studied in terms of the urease activity and interleukin (IL)-8 production capacity of different strains of H. pylori.. Forty-five patients infected with different strains of H. pylori (type I; 15, type II; 15 and type III; 15) were enrolled in the study. H. pylori was isolated from gastric mucosa and cultured in the presence of urea at pH 5.5 to evaluate urease activity. The capacity of different strains of H. pylori to induce IL-8 mRNA and IL-8 from a human gastric cancer cell line and human peripheral blood mononuclear cells was evaluated.. The urease activity of type II H. pylori[523 +/- 228 micro g of ammonia/dl/10(8) colony-forming units (CFU)/ml] was significantly lower than that of type I (1355 +/- 1369 micro g of ammonia/dl/10(8) CFU/ml) and type III (1442 +/- 2229 micro g of ammonia/dl/10(8) CFU/ml) (p <.05). Gastric cancer cells cocultured with type II H. pylori produced lower levels of IL-8 mRNA compared with type I and type III H. pylori. The levels of IL-8 were also significantly lower in cultures induced by type II H. pylori compared with those induced by type I and type III H. pylori. Peripheral blood mononuclear cells also produced lower levels of IL-8 when cocultured with type II compared with type I H. pylori.. These results indicate that both the lower level of urease activity and the low IL-8-inducing capacity of type II H. pylori might underlie the lower recurrence rate of gastritis caused by type II H. pylori.

Topics: Antigens, Bacterial; Bacterial Proteins; Cell Line; DNA Fingerprinting; DNA, Bacterial; Female; Gastritis; Gene Expression; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Recurrence; RNA, Messenger; Stomach Neoplasms; Urease

Helicobacter pylori colonizes the human stomach for decades unless pharmacologically eradicated. We hypothesized that this flagellated pathogen escapes immune clearance, in part, by avoiding detection by the flagellin receptor Toll-like receptor 5 (TLR5). In contrast to other gram-negative microbes, H. pylori did not release flagellin. Furthermore, recombinant H. pylori flagellin (FlaA) was significantly less potent (1000-fold) than Salmonella typhimurium flagellin in activating TLR5-mediated interleukin (IL)-8 secretion. TLR5 can mediate flagellin-induced IL-8 secretion via p38 mitogen-activated protein kinase signaling; however, compared with potent induction by S. typhimurium flagellin, H. pylori FlaA-dependent p38 activation was substantially attenuated. In addition, disruption of H. pylori flaA decreased motility but had no effect on H. pylori-induced IL-8 secretion, which indicates that H. pylori flagellin plays no role in activating epithelial orchestration of inflammation. We conclude that H. pylori evades TLR5-mediated detection, which may contribute to its long-term persistence in individual hosts.

Topics: Animals; Blotting, Western; Culture Media, Conditioned; Dogs; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Flagellin; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Membrane Glycoproteins; Mutagenesis, Insertional; Receptors, Cell Surface; Recombinant Proteins; Salmonella typhimurium; Stomach Diseases; Toll-Like Receptor 5; Toll-Like Receptors

Few studies have looked at the cytokine profile in gastric mucosa in children with Helicobacter pylori infection. This study investigated cytokines and their effects on histological abnormalities in the gastric mucosa of children with H. pylori infection.. The levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and IL-8 proteins were measured in biopsy specimens from the gastric antrum and corpus of children with H. pylori infection, and related to inflammatory cell infiltrations.. The antral and corporal mucosal levels of IFN-gamma and IL-8 proteins were significantly higher in children with H. pylori infection than in uninfected children, but there was no such difference in the levels of IL-4 protein. The antral mucosal level of IL-8 protein was significantly higher than the corporal mucosal level of IL-8 protein in the infected children. Inflammatory cell infiltration was significantly higher in the infected children than in the uninfected children, but there were no significant correlations between mucosal cytokine levels and inflammatory cell infiltrations.. The results suggest that the predominant Th1 cytokine response and enhanced IL-8 production in the mucosa may be involved in the gastric inflammation seen in children infected with H. pylori, as well as in adult patients.

Topics: Adolescent; Child; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interferon-gamma; Interleukin-4; Interleukin-8; Male; Neutrophils; Pyloric Antrum

Little information is currently available on the contribution of locally generated inflammatory and chemotactic cytokines to endothelial cell activation and subsequent neutrophil transendothelial migration in patients with Helicobacter pylori (H. pylori)-associated gastritis.. The contents of interleukin (IL)-1beta and IL-8 in the organ culture supernatants of antral mucosal tissues were measured with an enzyme-linked immunosorbent assay. The effects of the endogenous IL-1beta and IL-8 in mucosal tissues on neutrophil adherence and transendothelial migration were investigated using an experimental model of human umbilical vein endothelial cells (HUVEC).. The contents of IL-1beta and IL-8 in organ cultures of antral mucosal tissues were significantly higher in patients with H. pylori infection than in those without infection. The organ culture supernatants from H. pylori-positive patients induced the expression of intercellular adhesion molecule-1 mRNA in HUVEC with increased binding of neutrophils, and these stimulatory effects were inhibited when HUVEC were pretreated with a nuclear factor-kappaB inhibitor, MG-132. Moreover, neutrophil adherence to HUVEC induced by the supernatants decreased after preincubation with neutralizing anti-IL-1beta antibody. As compared with the supernatants from H. pylori-negative patients, the samples from H. pylori-positive patients exhibited a significantly higher chemotactic activity for neutrophils, which was inhibited almost completely by preincubation of the supernatants with anti-IL-8 antibody.. Locally generated IL-1beta and IL-8 could coordinate with each other during the process of neutrophil infiltration into the gastric mucosa in patients with H. pylori infection.

Topics: Adolescent; Adult; Aged; Cell Adhesion; Cell Movement; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Neutrophils; Pyloric Antrum

At the gastric cardia, the molecular mechanisms of inflammation and metaplasia are incompletely understood. Thus, the aim of this study was to determine the expression of TFF1, TFF2 and TFF3 at this site and correlate these data with Helicobacter pylori infection or gastro-esophageal reflux disease (GERD). In 27 patients without intestinal metaplasia at the cardia, endoscopic biopsies were obtained for histology and RT-PCR. TFF1 and TFF2 were expressed in all cardia samples. TFF3 expression was significantly more frequent at the cardia (n = 15/24) than in the corpus (n = 2/26). TFF3 expression at the cardia was mainly observed in GERD patients, and there was a clear tendency towards higher interleukin-8 (IL-8) transcription levels; whereas TFF3 expression was not correlated with the H. pylori status or to tumor necrosis factor-alpha (TNF-alpha) expression. The expression of TFF3 at the cardia may represent an adaptation to GERD and precede the development of Barrett's esophagus.

Topics: Barrett Esophagus; Cardia; Esophagogastric Junction; Gastroesophageal Reflux; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mucins; Muscle Proteins; Peptides; Proteins; Trefoil Factor-1; Trefoil Factor-2; Trefoil Factor-3; Tumor Necrosis Factor-alpha; Tumor Suppressor Proteins

Helicobacter pylori (H. pylori) is a non-invasive microorganism causing intense gastric mucosal inflammatory and immune reaction. H. pylori-induced gastric mucosal cytokine overproduction has been clearly documented previously. The stomach has a large surface area and continuous spill-over of locally produced cytokines into the blood stream is a possibility. There are few and conflicting data on circulatory proinflammatory cytokine levels in patients with H. pylori infection.. Forty-two dyspeptic patients were enrolled into the study. The presence of H. pylori infection was diagnosed with antral histopathologic examination. After overnight fasting; serum samples were obtained from each patient to determine circulating interleukin (IL)-6, IL-8 and tumor necrosis factor-alpha (TNF-alpha) levels.. H. pylori was shown in 30 cases using Giemsa stain in antral histopathologic evaluation. Twelve cases were negative for H. pylori staining. Both the age and sex distribution had an insignificant difference in both H pylori-positive and H. pylori-negative groups. The mean circulatory levels of IL-6, IL-8 and TNF-a in both groups were not different. The situation was same in respect to the serum levels of these cytokines and the degree of inflammation, H. pylori density and activation scores according to Sydney classification.. We could not show elevated circulatory levels of IL-6, IL-8 and TNF-alpha in H. pylori-infected cases. We believe that H. pylori-related cytokine activation become concentrated on gastric mucosa and this pathogen-induced local inflammatory cascade does not cause changes in circulatory levels of these cytokines. Moreover, there is no correlation between the levels of serum cytokines and Sydney parameters.

Topics: Adult; Cytokines; Dyspepsia; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

Helicobacter pylori infection leads to significant inflammations in the gastric mucosa, which is closely associated with development of gastric cancer. Heat shock protein 90 (HSP 90) has been revealed to be critical for intracellular signaling that participates in inflammatory response as well as carcinogenesis. In this study, we investigated a regulatory role of HSP 90 in H. pylori-induced IL-8 production. Our results showed that H. pylori stimulated significant phosphorylation of HSP 90 and the phosphorylation was diminished by administration of HSP 90 inhibitor, geldanamycin (GA). Treatment of GA completely inhibited H. pylori-induced IL-8 production due to deactivation of ERK1/2 and NF-kappaB. These results subsequently lead to inactivation of AP-1 and NF-kappaB, which are known to be major transcriptional factors of IL-8. Our data provide important insights that HSP 90 is involved as a crucial regulator in H. pylori-induced IL-8 production and its inhibitor could be potentially used for the inhibition of H. pylori-provoked inflammation.

Topics: Benzoquinones; Cell Line; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; HSP90 Heat-Shock Proteins; Humans; Interleukin-8; Lactams, Macrocyclic; NF-kappa B; Quinones; Transcription Factor AP-1; Transcriptional Activation

TLR4, a member of pattern recognition receptors, is the main receptor of LPS. MD-2 physically associates with TLR4 on the cell surface and confers LPS responsiveness. Helicobacter pylori LPS is one of the major virulence factors for induction of gastritis. We demonstrated in this study the role of MD-2 in TLR4-dependent signaling in H. pylori-associated gastritis. Gastric biopsy samples collected from patients with and without H. pylori infection and four gastric cancer cell lines were used for this study. TLR-4 and MD-2 expression in biopsy specimens and the cell lines was examined by using RT-PCR. Localization of TLR-4 in histological sections was evaluated by immunohistochemistry. For in vitro functional assays, we established stable transfectants of AGS cells expressing TLR4 and MD-2. Cellular distribution of TLR4 was examined by flow cytometry. NF-kappaB activation and activation of IL-8 and MD-2 promoters were assessed by reporter gene assay. H. pylori infection up-regulated the TLR4 and MD-2 expression in gastric mucosa. TLR4 staining was observed predominantly in epithelial cells, located in both the cytoplasm and at the apical surface. MD-2 transfection in AGS cells markedly increased cell surface expression of TLR4 and augmented the activation of NF-kappaB and IL-8 promoter upon stimulation with H. pylori LPS. Live H. pylori also stimulated transcriptional activation of MD-2. This study revealed that MD-2 expression is elevated in gastric epithelial cells during H. pylori infection, suggesting that the TLR4/MD-2 system is a potent receptor complex involved in the response to H. pylori LPS in the stomach.

Topics: Adult; Antigens, Surface; Female; Gastritis; Genes, Reporter; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lipopolysaccharides; Lymphocyte Antigen 96; Male; Membrane Glycoproteins; Middle Aged; NF-kappa B; Promoter Regions, Genetic; Receptors, Cell Surface; RNA, Messenger; Signal Transduction; Toll-Like Receptor 4; Toll-Like Receptors

The Helicobacter pylori cag pathogenicity island encodes a secretory system that translocates CagA into epithelial cells, where it becomes tyrosine phosphorylated and induces cytoskeletal rearrangements. Strains with more CagA tyrosine phosphorylation motifs are most closely associated with gastric cancer. Here we assess whether clinical strains can deliver CagA, whether strains with different numbers of CagA phosphorylation motifs have CagA phosphorylated to different degrees, and whether this induces different amounts of epithelial cytoskeletal change.. Forty-four H. pylori strains from South African patients, all cagA gene positive, were cocultured with the gastric adenocarcinoma cell line AGS. CagA expression and phosphorylation were determined by Western blot and interleukin-8 secretion by enzyme-linked immunosorbent assay. The cagA 3' variable regions of 22 strains were sequenced and shown to possess 3-6 phosphorylation motifs. These strains were used to quantify CagA phosphorylation and cytoskeletal rearrangements.. cagA genotype and typing of cag pathogenicity island genes were poorly predictive of phenotype. Thirty-four of 44 strains expressed CagA protein that could be delivered to and phosphorylated within AGS cells. Only these 34 strains induced interleukin-8 secretion from AGS cells. Among those strains, the number of CagA tyrosine phosphorylation motifs determined the degree of CagA phosphorylation and the level of biologic activity in terms of degree and extent of AGS cell elongation.. H. pylori strains that deliver CagA with more phosphorylation motifs induce higher levels of CagA phosphorylation in epithelial cells, induce more cytoskeletal changes, and are more likely to be associated with gastric cancer.

Topics: Amino Acid Sequence; Antigens, Bacterial; Bacterial Proteins; Cells, Cultured; Cytoskeleton; Epithelial Cells; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Molecular Sequence Data; Peptide Fragments; Phosphorylation; Polymerase Chain Reaction; Tyrosine; Virulence

Helicobacter pylori infection of the stomach is commonly associated with infiltration of neutrophils. Gastric epithelial cells are recognized as central mediators of tissue responses to this organism. The aim of the present study was to ascertain patterns of production of three neutrophil chemoattractant chemokines following infection of gastric epithelial cells with H. pylori in vitro.. Two gastric cancer-derived epithelial cell lines were infected with H. pylori organisms of previously defined cagE and cagA status for periods of up to 24 h. The production of three chemokines (interleukin [IL]-8, epithelial neutrophil activating protein [ENA]-78 and growth-related oncogene [GRO]-alpha) over this time was measured in culture supernatants using immunoassays.. Both IL-8 and GRO-alpha were produced by both AGS and KATO-III cells in response to H. pylori infection, and in a cag PAI-dependent manner. ENA-78, however, was not increased following H. pylori infection.. GRO-alpha is expressed by epithelial cells following H. pylori infection along with IL-8. Both may contribute to neutrophilic infiltration present in gastric mucosa associated with H. pylori infection. In contrast, H. pylori infection does not lead to an increased synthesis of ENA-78, suggesting that this may not be as important in vivo.

Topics: Analysis of Variance; Cell Line; Chemokine CXCL1; Chemokine CXCL5; Chemokines, CXC; Enzyme-Linked Immunosorbent Assay; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; In Vitro Techniques; Intercellular Signaling Peptides and Proteins; Interleukin-8; Tumor Cells, Cultured

Helicobacter pylori-induced mucosal inflammation results in high production of interleukin 17 (IL-17), a potent inducer of IL-8 in gastric epithelial cells. The aim of this study was to investigate signaling pathways by which IL-17 regulates IL-8 production in human gastric epithelial cells. Activation of mitogen-activated protein (MAP) kinases in both IL-17-stimulated MKN28 cells and epithelial cells isolated from H. pylori-colonized gastric mucosa was assessed by Western blotting. In IL-17-stimulated MKN28 cells the activation of activatior protein 1 (AP-1), nuclear factor (NF)-IL-6, and NF-kappaB was also assessed by electrophoretic mobility shift assay. IL-8 production was evaluated by reverse transcription-PCR and enzyme-linked immunosorbent assay (ELISA) both for IL-17-stimulated MKN28 cells treated with specific MAP kinase inhibitors and gastric biopsy cultures treated with a neutralizing IL-17 antibody. Serum from H. pylori-infected patients was tested for immunoglobulin G response to CagA by ELISA. Treatment of MKN28 cells with IL-17 caused activation of extracellular signal-regulated protein kinase 1/2 (ERK 1/2) but not other MAP kinases and had the downstream effects of AP-1 and NF-kappaB activation and IL-8 synthesis. Blocking ERK 1/2 activity inhibited AP-1-mediated, but not NF-kappaB-mediated, IL-8 induction. Enhanced activation of ERK 1/2 was seen in gastric epithelial cells isolated from H. pylori-infected patients in comparison to uninfected controls, and this was associated with high IL-8. These effects were even more pronounced in patients seropositive for CagA than in seronegative ones. In gastric biopsy cultures, the addition of a neutralizing IL-17 antibody decreased ERK 1/2 activation, thus resulting in a significant inhibition of IL-8. In H. pylori-colonized gastric epithelial cells, IL-17-induced IL-8 synthesis is associated with and depends at least in part on the activation of ERK 1/2 MAP kinase.

Topics: Adult; Aged; Cell Line; Cells, Cultured; Enzyme Activation; Epithelial Cells; Female; Gastric Mucosa; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases

Thrombotic microangiopathy (TMA) has attracted attention as a complication of bone marrow transplantation (BMT). The association of Helicobacter pylori (H. pylori) with thrombotic thrombocytopenic purpura and hemolytic uremic syndrome (TTP/HUS) after BMT was studied. Among 74 consecutive patients undergoing transplantation, six developed TTP/HUS (the TTP/HUS group) and 68 did not (controls). These six patients were compared with the other 68 patients to investigate differences of the IL-12 and 8 levels, H. pylori and various clinical characteristics. The patients who developed TTP/HUS seemed not apparently different from those who did not in background characteristics, except that they had a significantly higher H. pylori-positive rate (p < 0.05). In the TTP/HUS group, however, the levels of interleukin-12 and interleukin-8 increased significantly during the leukocyte recovery after BMT and at the onset of TTP/HUS, respectively, to 45.8 +/- 57.6 pg/mL and 274.8 +/- 65.9 pg/mL (p < 0.05 for both), when compared with their levels of 5.0 pg/mL in the control group. Thus, H. pylori may play a role in the pathogenesis of TTP/HUS after BMT, with cytokines (interleukin-8 and interleukin-12) also being involved.

Topics: Adolescent; Adult; Bone Marrow Transplantation; Colony Count, Microbial; Female; Helicobacter Infections; Helicobacter pylori; Hemolytic-Uremic Syndrome; Humans; Interleukin-12; Interleukin-8; Leukocytes; Male; Middle Aged; Purpura, Thrombotic Thrombocytopenic; Retrospective Studies; Risk Factors; Transplantation Conditioning

Topics: Animals; Female; Helicobacter Infections; Humans; Interleukin-6; Interleukin-8; Male; Stomach Neoplasms; Tumor Necrosis Factor-alpha

Topics: Apoptosis; DNA; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interferon-gamma; Interleukin-6; Interleukin-8; RNA, Messenger; Stomach Ulcer; Tumor Necrosis Factor-alpha

Helicobacter pylori (H. pylori) infection causes various gastric diseases, among them H. pylori-associated gastritis characterized by diffuse redness of the gastric mucosa. The haemoglobin index (IHb) of the fundic mucosa is an objective parameter of the extent of mucosal redness, but it is unclear whether or not IHb can be used as a diagnostic marker for H. pylori infection. The purpose of this investigation was to evaluate the correlations between IHb of the fundic mucosa and H. pylori infection, inflammatory cell infiltration, and inflammatory mediator production.. IHb of the fundic mucosa was measured in 108 patients with various gastric diseases (group 1), and values were compared between H. pylori-positive and H. pylori-negative patients. Fifteen patients with H. pylori infection from group 1 underwent H. pylori eradication therapy and IHb was measured before and after treatment. Both IHb and inflammatory cell infiltration were assessed in 61 patients (group 2). In 31 patients from group 2, the expression of interleukin (IL)-8 and inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was assayed in gastric biopsy specimens by the reverse transcription-polymerase chain reaction (RT-PCR).. IHb levels were significantly higher in H. pylori-positive patients than in H. pylori-negative patients (P < 0.001). IHb was decreased at one month after the eradication of H. pylori (P < 0.001). IHb was higher in patients with infiltration by both mononuclear cells and neutrophils (P < 0.001). There was a significant correlation between the IHb level and the expression of IL-8 mRNA (P < 0.001), as well as between IHb and iNOS mRNA expression (P < 0.05).. There were significant correlations between IHb of the gastric mucosa and H. pylori infection, inflammatory cell infiltration, and IL-8/iNOS mRNA expression, suggesting that IHb is a reliable marker of H. pylori infection for use during follow-up endoscopy after H. pylori eradication therapy.

Topics: Biomarkers; Gastric Mucosa; Gastritis; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Hemoglobins; Humans; Inflammation; Interleukin-8; Neutrophils; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Regional Blood Flow; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

In Helicobacter pylori gastritis, neutrophil activation and migration, which play central roles in the pathogenesis of the disease, are regulated by the neutrophil attractant chemokines interleukin 8 (IL-8) and Groalpha, whose secretion is induced by H. pylori. However, the modulation of the corresponding chemokine receptors CXCR1 and CXCR2 on human neutrophils under the influence of H. pylori has not been investigated. Incubation of neutrophils with cag(+) and cag deletion H. pylori strains resulted in a complete downregulation of the CXCR1 and the CXCR2 receptors after 0.5 h, as tested by fluorescence-activated cell sorter analysis, independent of the cag status. Downregulation of CXCR1 and CXCR2 seems to occur via receptor internalization and rapid degradation, as shown by confocal microscopy and immunoblotting. Neither the proinflammatory cytokines IL-8 and tumor necrosis factor alpha produced by the neutrophils themselves nor H. pylori lipopolysaccharide, which are the known regulators of these two chemokine receptors, was responsible for the downregulation. Reverse transcription-PCR analysis showed that CXCR1 and CXCR2 mRNAs of neutrophils were reduced at a later time than the CXCR1 and CXCR2 proteins. Moreover, cag(+) H. pylori strains induced significantly stronger downregulation of CXCR1 and CXCR2 mRNAs than the cag deletion mutant. Therefore, receptor protein and mRNA downregulation seem to be mediated by two independent mechanisms. Data obtained by immunohistochemistry suggested that downmodulation of CXCR1 and CXCR2 on neutrophils may also occur in vivo in the human stomach during H. pylori infection. Downregulation of CXCR1 and CXCR2 expression on neutrophils in H. pylori infection by H. pylori itself may represent a new mechanism of modulating neutrophil migration and activation in the gastric mucosa.

Topics: Down-Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lipopolysaccharides; Neutrophils; Receptors, Interleukin-8A; Receptors, Interleukin-8B; RNA, Messenger; Tumor Necrosis Factor-alpha

Topics: Esophagoscopy; Esophagus; Gastroesophageal Reflux; Helicobacter Infections; Humans; Immunohistochemistry; Interleukin-8; Mucous Membrane; NF-kappa B; RNA, Messenger

Elevated serum gastrin and a low pepsinogen A/C ratio are well-recognized markers for atrophic body gastritis (ABG). We have shown that the presence of body atrophy is also associated with elevated serum pro-inflammatory cytokines. This study tested the hypothesis that serum cytokines provide additional information to gastrin and pepsinogens in screening for ABG.. Two hundred and twenty-six consecutive patients were investigated on referral for upper gastrointestinal endoscopy: 150 were patients with gastro-oesophageal reflux disease, receiving acid inhibitory medication either with proton pump inhibitors (n = 113) or with histamine2-receptor antagonists (n = 37), and 76 were nontreated controls, who had normal endoscopic findings. Gastric mucosal biopsies were sampled for histological examination (Sydney classification). Serum samples were analyzed for gastrin, chromogranin A (CgA), and pepsinogens A and C by RIA, and for the interleukins (IL)-1beta, IL-6, and IL-8 by ELISA.. Subjects with ABG had significantly higher serum gastrin (P < 0.01) and serum CgA (P < 0.01) levels and significantly lower pepsinogen A/C ratios (P < 0.001) than those without ABG. Additionally, serum IL-1beta, IL-6 and, especially, IL-8 levels were significantly higher in the subjects with than in those without ABG (P < 0.0001, for all cytokines). To optimize the detection of body atrophy we defined the ABG index: the ratio between the simultaneously measured IL-8 and pepsinogen A/C. The area under the ROC curve for the ABG index was significantly greater than that for serum gastrin and for serum pepsinogen A/C alone (0.91 +/- 0.029 vs. 0.72 +/- 0.042, and vs. 0.83 +/- 0.031, P = 0.018 and P = 0.049). Using the ABG index at a cut-off value of 1.8 pg mL-1, 91% of the cases were classified correctly.. The ratio between serum IL-8 and pepsinogen A/C accurately predicts the presence of ABG. We therefore propose the ABG index as a noninvasive screening test for ABG in population-based studies.

Topics: Adult; Aged; Biomarkers; Female; Gastrins; Gastritis, Atrophic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Male; Mass Screening; Middle Aged; Pepsinogen A; Pepsinogen C; Pepsinogens; ROC Curve; Sensitivity and Specificity

Oxygen radicals are important regulators in Helicobacter pylori-induced gastric ulceration and carcinogenesis. IL-8 may be regulated by oxidant-sensitive transcription factors, NF-kappaB, and AP-1. The present study aims to investigate whether H. pylori-induced IL-8 expression is regulated by NF-kappaB and AP-1 in gastric epithelial AGS cells and whether this transcriptional regulation of IL-8 is inhibited by N-acetylcysteine (NAC). As a result, H. pylori induced the expression of mRNA and protein for IL-8 via activation of NF-kappaB and AP-1. NF-kappaB activation accompanied by a decrease in I-kappaBalpha and activated AP-1 complex was a c-jun/c-fos heterodimer in H. pylori-infected AGS cells. NAC inhibited H. pylori-induced activation of transcription factors and IL-8 expression in AGS cells. In conclusion, oxygen radicals induce the activation of NF-kappaB and AP-1 and IL-8 expression. Antioxidants such as NAC might be useful anti-inflammatory agents by inhibiting activation of transcription factors and decreasing IL-8 production in H. pylori-induced gastric inflammation.

Topics: Analysis of Variance; Base Sequence; Blotting, Northern; Blotting, Western; Cells, Cultured; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Molecular Sequence Data; NF-kappa B; Polymerase Chain Reaction; Probability; Reactive Oxygen Species; RNA, Messenger; Sensitivity and Specificity; Stomach Ulcer; Transcription Factor AP-1

Chemokines play a key role in the pathogenesis of various inflammatory conditions. However, there is little information on their profile in reflux esophagitis (RE). We sought to study esophageal mucosa levels of chemokines in RE.. A total of 32 outpatients with RE and 13 normal controls were studied. Endoscopic severity of RE was classified according to the Los Angeles grading system. Paired biopsy specimens were taken from the esophagus 3 cm above the gastroesophageal junction; one biopsy was snap frozen for measurement of mucosal levels of interleukin 8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), regulated on activation normal T-cell expressed and presumably secreted (RANTES), and IL-1 beta by enzyme linked immunosorbent assays, while the other was formalin-fixed for histopathological evaluation.. IL-8, MCP-1, and RANTES levels were significantly higher in esophageal mucosa of RE patients than those of the controls. IL-8 levels correlated significantly with the endoscopic severity of RE. Basal zone hyperplasia and papillary elongation, histopathological hallmarks of RE, were both associated with higher levels of IL-8 and MCP-1. The presence of intraepithelial neutrophils and eosinophils, which also indicate RE, was associated with high levels of IL-8 and RANTES, respectively. There were no significant differences in IL-1 beta levels between the RE and control groups, but IL-1 beta levels correlated significantly with the IL-8 production. Again, the IL-8 levels were significantly decreased after lansoprazole treatment.. Our results indicate that chemokines produced locally in the esophageal mucosa may be involved in the development and progression of RE.

Topics: Adult; Aged; Aged, 80 and over; Biopsy; Chemokine CCL2; Chemokine CCL5; Chemokines; Enzyme-Linked Immunosorbent Assay; Esophagitis, Peptic; Esophagus; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Mucous Membrane; Severity of Illness Index

Helicobacter pylori induces inflammation of gastric mucosa regulated by several interleukins. This study examined associations between anti-Helicobacter pylori immunoglobulin G antibody seropositivity and functional polymorphisms of interleukin-8 T-251 A and interleukin-10 T-819C.. The subjects were 454 health check-up examinees (126 males and 328 females) without a history of cancer, aged 35-85 years, residing in Nagoya, Japan. After written informed consent was obtained individually, residual blood was anonymously applied for anti-Helicobacter pylori immunoglobulin G antibody testing and genotyping by the polymerase chain reaction with confronting two-pair primers.. The genotype frequency of interleukin-8 T-251 A was 52.2% for TT, 39.5% for TA, and 8.3% for AA, and that of interleukin-10 T-819C was 49.5% for TT, 39.9% for TC and 10.6% for CC. Although the differences in the positive rates among the genotypes were not marked, 115 individuals with interleukin-8-251TT (low expression genotype) and interleukin-10-819TT (high expression genotype) had a higher rate (63.5%) than the others (52.0%). Relative to the combination of interleukin-8-251TT and interleukin-10-819TT, the sex-age-adjusted odds ratio for those with the other combinations was 0.62 (95% confidence interval, 0.39-0.98). The adjusted odds ratio among 65 current smokers was 0.13 (0.03-0.61).. The observed association suggests that individuals with interleukin-8-251TT and interleukin-10-819TT, a combination presumably causing mild inflammation, have a higher probability of the continuing Helicobacter pylori infection, especially among current smokers.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Bacterial; Female; Genetic Predisposition to Disease; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged

The evaluation of eosinophil cationic protein (ECP) concentration--one of late allergy reaction markers was performed in serum of children with food allergy and children with food allergy and H. pylori or Giardia lamblia infection of the gastrointestinal tract. The ECP values were referred to the characteristics of histopathological changes in gastric mucosa and to the values of cytokines (IL-4, IL-5, IL-8) determined in biopsy specimens of gastric mucosa from these patients. The studies indicate that the exclusive evaluation of ECP concentration in serum does not reflect unequivocally the severity of pathological changes of gastric mucosa in children with food allergy.

Topics: Adolescent; Blood Proteins; Case-Control Studies; Child; Enzyme-Linked Immunosorbent Assay; Eosinophil Granule Proteins; Female; Food Hypersensitivity; Gastric Mucosa; Giardiasis; Helicobacter Infections; Humans; Inflammation Mediators; Interleukin-4; Interleukin-5; Interleukin-8; Intestinal Mucosa; Male; Ribonucleases

Billroth I or II reconstruction after distal gastrectomy often is associated with inflammation in the gastric remnant. We sought to determine which reconstructive procedure was most effective in preventing such remnant gastritis. Patients undergoing curative distal gastrectomy for cancer ( n = 82) were classified as group A (Roux-en- Y, n = 22); group B (Billroth I, n = 40); or group C (Billroth II, n = 20). Interleukin (IL)-8 concentrations in gastric mucosa were measured 3 months after surgery. In the absence of Helicobacter pylori infection, IL-8 concentrations were 13, 56, and 87 pg/mg protein in groups A, B, and C, respectively ( p < 0.05). In the presence of H. pylori infection, IL-8 concentrations were 61, 161, and 234 pg/mg protein in groups A, B, and C ( p < 0.01). Roux-en- Y reconstruction is better able to prevent remnant gastritis than either the Billroth I or II procedure as judged from IL-8 concentrations in gastric remnant mucosa.

Topics: Adult; Aged; Aged, 80 and over; Duodenogastric Reflux; Female; Gastrectomy; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Stomach Neoplasms

Isolates of Helicobacter pylori from dyspeptic patients in England and South Africa were tested for ability to induce interleukin-8 (IL-8) in gastric cells. All isolates were cagA-positive, which was used as a marker for the presence of the cag pathogenicity island. The aims were to determine if activities were related to diversity within cagE (HP0544), a locus encoding a key component in the Type IV secretion system, and if disease severity might be linked to a combination of strain features. We found that isolates were heterogeneous in ability to induce IL-8 activity with the 23 positive isolates (59%) showing activities ranging from 260 to 3200 pg ml(-1). The cagE locus was detected in most isolates and RFLP analysis of a 1.52-kb internal fragment showed interstrain diversity with 12 combined (MboI/NlaIII) types. Most cagE genotypes were not associated with IL-8 induction, however two genotypes were found only in IL-8-inducing strains and one genotype was associated with lack of IL-8 induction. IL-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. Although we found a weak association between cagE type and the ability to induce IL-8, our results imply that gastric cell factors or bacterial factors other than vacA, cagA and cagE are involved in the induction of IL-8 and the development of severe gastric disease.

Topics: Antigens, Bacterial; Bacterial Proteins; Dyspepsia; Epithelial Cells; Gastric Mucosa; Gastrointestinal Diseases; Genetic Variation; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Phosphorylation; Severity of Illness Index; Tyrosine

Helicobacter pylori is an important pathogen in gastroduodenal inflammation and ulceration. Several mechanisms have been proposed to explain its role. We studied the cytokine production patterns in situ in gastric mucosal biopsies from H. pylori-positive and H. pylori-negative patients with dyspepsia. Immunohistochemistry with monoclonal antibodies was used. The study showed enhanced expression of interleukin (IL) -8, IL-10 and interferon-gamma (IFN-gamma) in H. pylori infection and a significant association was found between these cytokines and the following parameters: bacteria load, chronic inflammation and activity. These parameters were significantly correlated with the cell markers CD19 and CD56. The study indicates a dual effect of H. pylori on the Th1 response, i.e. a stimulation of the response verified by increased IFN-gamma and a feed-back verified by an increase of the counterinflammatory IL-10, which may dampen the inflammatory and cytotoxic effect of the Th1 response. Furthermore, the study confirms the connection between increase of IL-8 and inflammatory activity in gastric mucosa in H. pylori infection.

Topics: Adult; Aged; Aged, 80 and over; Chronic Disease; Colony Count, Microbial; Cytokines; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Inflammation; Interferon-gamma; Interleukin-10; Interleukin-8; Male; Middle Aged; Peptic Ulcer

To date only a few Helicobacter pylori strains have been demonstrated to colonise Mongolian gerbils successfully. The aim of this study was to establish stable colonisation of Chinese strains of H. pylori in gerbils. Fresh clinical isolates from Chinese patients were inoculated into gerbils. At 4-6 weeks post inoculation, infection status was evaluated by culture, biopsy urease test and pathology. Sequencing of glmM and random amplified polymorphic DNA (RAPD) fingerprinting of DNA from cultured H. pylori were used to evaluate the genetic identity of pre-inoculated and post-inoculated strains. The ability of pre- and post-inoculated strains to stimulate interleukin-8 transcription in L5F11 gastric epithelial cells was analysed. Three of five clinical isolates colonised gerbils. The three pre- and post-inoculation strains had identical glmM sequences and RAPD profiles, and stimulated luciferase secretion from L5F11 epithelial cells. The strain that caused severe pathological changes was selected for repeat infection to prove reproducible and stable colonisation. The cagA+ strain 42GX gave stable colonisation in the gerbil and induced severe gastritis.

Topics: Animals; Base Sequence; Cell Line; Disease Models, Animal; Gastric Mucosa; Gastritis; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Molecular Sequence Data; Phosphoglucomutase; Random Amplified Polymorphic DNA Technique; Sequence Analysis, DNA

There is no direct evidence for an animal model of Helicobacter pylori induced duodenal ulcer.. In this study we evaluated the roles of bacterial strain and age of experimental animals in induction of duodenitis and duodenal ulcer in Mongolian gerbils after H pylori infection.. Specific pathogen free Mongolian gerbils were inoculated orally with three bacterial strains (H pylori ATCC 43504, TN2GF4, and K-6, a clinical isolate from a patient with gastric cancer in our clinic). These strains have both the cagA gene and VacA. Five week old gerbils were used to emulate prematurity infection and 14 week old animals were used as mature test subjects. Animals were observed for 12 weeks after inoculation. Interleukin 8 (IL-8) production in gastric epithelial cells (MKN74) after coculture with the H pylori strains was measured by ELISA.. Gastritis and gastric ulcers were found in all gerbils infected with the three strains. However, duodenitis and gastric metaplasia were seen more frequently in gerbils infected with TN2GF4 and K-6 strains than in the ATCC 43504 infected or control groups (p<0.05). Superficial duodenal ulcers with severe duodenitis and gastric metaplasia were found in two gerbils inoculated at 14 weeks with the TN2GF4 strain but none at five weeks. The TN2GF4 strain stimulated significantly higher levels of IL-8 than ATCC 43504 and K6 strains (p=0.0039).. When injected into adult Mongolian gerbils, a specific strain (TN2GF4) of H pylori can induce duodenitis with gastric metaplasia and superficial duodenal ulcers. Induction of duodenal ulcer in an animal model fulfills the requirements of Koch's postulates for establishing a role for H pylori as a causative agent.

Topics: Age Factors; Animals; Coculture Techniques; Disease Models, Animal; Duodenal Ulcer; Duodenitis; Gastric Mucosa; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Male; Metaplasia

To investigate the role of nuclear factor kappaB (NF-kappaB) in the gastric inflammation induced by Helicobacter pylori (H. pylori).. SGC-7901 was transfected with IkappaB (NF-kappaB inhibitor gene) by electroporation, (beta)lacZ activity assay was used to examine transfected efficacy. Expression of IkappaB was assessed by Western-blot. Different concentration of live and heat-killed Hp (ATCC 43504) and supernatant of liquid culture were cocultured with SGC7901-IkappaB and its negative control SGC7901- neo. Activation of intracellular NF-kappaB was examined by electrophoretic mobility shift analyses (EMSA) and luciferase report gene assay at different time point. IL-8 levels were measured by ELISA at different time point.. IL-8 release was evident 4 hours after infection of SGC-7901-neo with H. pylori, and this effect was dose-time dependent. SGC-7901-IkappaB in which NF-kappaB has not been activated could not secret IL-8 after infection with H. pylori.. Secretion of IL-8 by gastric epithelial cell upon H pylori infection is dependent on activation of NF-kappaB.

Topics: Cell Line, Tumor; Electroporation; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; RNA, Messenger; Stomach Neoplasms

To investigate the relation between serum level of interleukin-8 (IL-8), nitrogen monoxide (NO) and Helicobacter pylori (HP) infection, as well as the possible molecular mechanism of HP infection causing the imbalance of apoptosis and proliferation in gastric epithelial cells, which may lead to oncogenesis in stomach.. Serum IL-8 level was detected with enzyme-linked immunosorbent assay (ELISA). Serum NO was detected with chrome reduction method using plated copper.. Serum level of IL-8 were 22.50 +/- 1.87 pg/ml in the normal tissue, 34.99 +/- 7.89 pg/ml in superficial gastritis, 65.27 +/- 10.60 pg/ml in atrophic gastritis and 94.84 +/- 11.09 pg/ml in gastric cancer (P < 0.01). Serum level of NO in the atrophic gastritis group (39.93 +/- 5.43 micromol/L) was significantly higher than that in the gastric cancer group (37.02 +/- 4.13 micromol/L) (P < 0.05). The differences in the other groups were not significant. IL-8 and NO levels in the HP(+) group were significantly higher than those in the HP(-) group (77.30 +/- 20.92 pg/ml vs 39.16 +/- 14.40 pg/ml, P < 0.01; 39.77 +/- 5.57 micromol/L vs 35.35 +/- 5.24 micromol/L, P < 0.01). Serum levels of IL-8 and NO in the cytotoxin-associated gene A protein (CagA)(+)HP group were significantly higher than those in the CagA(-)HP group (83.45 +/- 16.92 pg/ml vs 66.24 +/- 23.21 pg/ml, P < 0.01; 40.97 +/- 4.59 micromol/L vs 37.62 +/- 6.58 micromol/L, P < 0.05). Serum levels of IL-8 and NO showed positive correlation between superficial gastritis and atrophic gastritis groups (r = 0.881, r = 0.995), whereas no correlation was found in the normal or gastric cancer groups.. Serum levels of IL-8 and NO are correlated with CagA(+)HP strain infection. Combined detection of serum level of IL-8, NO and HP-CagA will contribute to the early diagnosis of precancerous lesion in the stomach.

Topics: Antigens, Bacterial; Bacterial Proteins; Cell Proliferation; Early Detection of Cancer; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Nitric Oxide; Stomach Neoplasms

Clarithromycin (CAM)-resistant Helicobacter pylori sometimes offers serious problems with eradication by antibiotics. The aim of this study was to determine whether a probiotic can be an alternative therapy in CAM-resistant Hp infection.. The effects of Lactobacillus gasseri (strain OLL2716) on the growth of CAM-susceptible and CAM-resistant H. pylori and interleukin (IL)-8 production provoked by these strains were examined by in vitro experiments. Moreover, mice were infected with these CAM-susceptible or CAM-resistant H. pylori, and were treated with CAM or L. gasseri.. In vitro experiments demonstrated that L. gasseri inhibited the growth of H. pylori and suppressed H. pylori-associated IL-8 production. Such effects were noted in CAM-resistant and CAM-susceptible H. pylori. Similarly, in an in vivo model of H. pylori infection, H. pylori colonization was significantly decreased by L. gasseri.. Therefore, L. gasseri was found to act as a probiotic in CAM-resistant H. pylori infection.

Topics: Animals; Anti-Bacterial Agents; Clarithromycin; Drug Resistance, Bacterial; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Lactobacillus; Male; Mice; Mice, Inbred BALB C; Microbial Sensitivity Tests; Probiotics

and aims: Matrix metalloproteinases (MMPs) are endopeptidases with roles in extracellular matrix remodelling, cell proliferation, and inflammatory processes. We showed previously that Helicobacter pylori infection of human gastric adenocarcinoma (AGS) cells increased epithelial secretion of epithelial MMP-1 and MMP-3 and bacterial secretion of MMP-3-like activity. In the present study, we sought to characterise the role of interleukin (IL)-1beta in H pylori induced secretion of epithelial MMPs.. AGS cells were treated with H pylori and/or IL-1beta. Comparable IL-8 secretory responses (approximately 1700 ng/ml) measured by ELISA were induced by 2.0 ng/ml IL-1beta and by H pylori at a multiplicity of infection (MOI) of 50. The same IL-1beta and H pylori concentrations induced comparable increases in AGS cell caseinolytic activity at 60 kDa. MMP-3 monoclonal antibody immunoblots of AGS cell conditioned media detected immunoreactive bands at 71 kDa and 56 kDa. H pylori (MOI=50-100) induced dose dependent increases in both bands whereas IL-1beta (0.2-2 ng/ml) induced dose dependent increases only in the 71 kDa band, which was identified as a MMP-3/TIMP-3 (tissue inhibitor of metalloproteinases 3) heterodimer. AGS/H pylori conditioned media expressed 24 times more MMP-3 activity than AGS/IL-1beta conditioned media. There was a strong interaction between IL-1beta and H pylori on MMP-3 secretion.. We conclude that IL-1beta induces gastric epithelial cell MMP-3 secretion, contributing to epithelial tissue destruction during H pylori infection. However, other bacterial/host factors are needed to mediate the full gastric epithelial cell MMP-3 secretory response induced by H pylori infection.

Topics: Culture Media, Conditioned; Enzyme Activation; Gastrointestinal Neoplasms; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Intestinal Mucosa; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Receptors, Interleukin-1; Tissue Inhibitor of Metalloproteinase-3; Tumor Cells, Cultured

Helicobacter pylori infection, which is always associated with gastritis, can progress to ulceration or malignancy. The diversity in clinical outcomes is partly attributed to the expression of virulence factors and adhesins by H. pylori. However, H. pylori may not have to adhere to the epithelium to cause gastritis. We hypothesize that outer membrane vesicles (OMV), which are constantly shed from the surface of H. pylori, play a role as independent activators of host cell responses. In this study, we found that low doses of OMV from cag PAI+ toxigenic and cag PAI- nontoxigenic strains increased proliferation of AGS gastric epithelial cells. At higher doses, we detected growth arrest, increased toxicity, and interleukin-8 (IL-8) production. The only strain differences detected were vacuolation with the toxigenic strain and higher levels of IL-8 production with OMV from the cag PAI- nontoxigenic strain. In summary, we suggest that constitutively shed OMV play a role in promoting the low-grade gastritis associated with H. pylori infection.

Topics: Cell Division; Cell Line; Cell Membrane; Epithelial Cells; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Stomach; Virulence

Migration of neutrophil across the intestinal epithelium is a structural hallmark of active intestinal diseases. One of the characteristic histologic features of Helicobacter pylori (H. pylori) infection is neutrophil infiltration of the gastric mucosa. This study was aimed to establish an in vitro model system of transepithelial migration of neutrophil by H. pylori.. Caco-2 cells were grown on polycarbonate membrane. Confluence and integrity were assessed by measuring the transepithelial electrical resistance (TER) and paracellular permeability of macromolecular marker. IL-8 mRNA expression was assessed after H. pylori infection by quantitative RT-PCR. Isolated neutrophils and H. pylori were applied to the basolateral and apical compartment in separate. Also neutrophil transmigration was assessed by light and electron microscopy.. Paracellular leakage of 3[H]-mannitol was negligible over TER 166 Omega x cm2. IL-8 mRNA expression increased after H. pylori infection. Neutrophil transmigration was increased by adding of N-formyl-methionyl-leucyl-phenylalanine or H. pylori.. This in vitro model system could be applied to the study of pathogenesis of H. pylori infection. A further study of quantification of transmigrated neutrophils is demanded to reach a confirmation.

Topics: Caco-2 Cells; Cell Movement; Cells, Cultured; Electric Impedance; Epithelium; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Membranes, Artificial; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Permeability; Polycarboxylate Cement; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Few reports exist on inflammation and interleukin (IL)-8 response in H. pylori-infected children. The aim of this study was to determine the intensity of inflammation, density of colonization and magnitude of IL-8 response in children with and without H. pylori infection.. We studied 45 children with dyspeptic symptoms, 21 infected with H. pylori and 24 without infection. Antrum and corpus gastric biopsies were obtained and studied for H. pylori infection with an immunofluorescence technique and for IL-8 with an immunohistochemical assay. Biopsy specimens were stained with hematoxilin and eosin and gastritis was graded according to the Sydney system. The magnitudes of the IL-8 response and H. pylori colonization were estimated microscopically with image analyzer software.. In H. pylori-infected children, mild mono-nuclear cell infiltration was found in 50%, and no neutrophils in 40% of cases. In the antrum but not in the corpus, the intensity of colonization correlated with neutrophil and mononuclear cell infiltration. The IL-8 response was significantly higher in the antrum (p <.05) and corpus (p <.02) of infected children, and was localized mainly in the surface and crypts of the epithelium. No correlation was found between the magnitude of the IL-8 response and the infiltration of either neutrophil or mononuclear cells.. In H. pylori-infected children, poor mononuclear and neutrophil infiltration was observed. Infection was associated with a higher IL-8 response by gastric epithelial cells. The density of colonization but not the IL-8 response correlated with neutrophil cell infiltration.

Topics: Adolescent; Child; Dyspepsia; Epithelial Cells; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Leukocytes, Mononuclear; Male; Neutrophil Infiltration

Little has been known about the pathogenesis of non-erosive reflux disease (NERD). Recent studies have implicated interleukin 8 (IL-8) in the development and progression of gastroesophogeal reflux disease (GERD). The purpose of this study was to determine IL-8 RNA expression levels in NERD patients with or without subtle mucosal changes.. We studied 26 patients with NERD and 13 asymptomatic controls. Biopsy sample was taken from the esophagus 3 cm above the gastroesophageal junction and snap frozen for measurement of IL-8 mRNA levels by real-time quantitative polymerase chain reaction (PCR). We also examined mRNA expression of IL-8 receptors, CXCR-1 and -2 by reverse transcriptase PCR. The patients were endoscopically classified into grade M (mucosal color changes without visible mucosal break) and N (neither minimal involvement nor mucosal break) of the modified Los Angeles classification.. The relative IL-8 mRNA expression levels were significantly higher in esophageal mucosa of NERD patients than those in esophageal mucosa of the controls. There was a significant difference in IL-8 mRNA levels between grades M and N. The CXCR-1 and -2 mRNAs were constitutively expressed in esophageal mucosa.. Our results suggest that high IL-8 levels in esophageal mucosa may be involved in the pathogenesis of NERD through interaction with its receptors. NERD seems to be composed of a heterogeneous population in terms of not only endoscopically minimal involvement but also immune and inflammatory processes.

Topics: Adult; Aged; Alcohol Drinking; Base Sequence; DNA Primers; Endoscopy, Digestive System; Female; Gastroesophageal Reflux; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Hernia, Hiatal; Humans; Interleukin-8; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; RNA, Messenger; Smoking

Numerous studies have shown an association between Helicobacter pylori (Hp) infection and gastric cancer (GC).. This study was designed to determine the role of cytotoxin-associated gene A (CagA)-positive Hp infection, serum amidated gastrins and their precursor, progastrin, gastric acidity and serum pepsinogen I (PG-I) levels in gastric cancerogenesis in 74 cancer patients and in 77 age- and gender-matched controls. Serum IgG antibodies to Hp and CagA and levels of IL-8 and PG-I were measured by ELISA, while progastrin and amidated gastrin by specific radioimmunoassay.. The overall Hp and CagA seropositivity in GC patients were significantly higher (82 and 60%) than in matched controls (61 and 27%, respectively). Progastrin and amidated gastrin levels over their cutoff points (122 and 32 pM, respectively) were found in a significantly larger number of GC (59.4 and 44.5%) than in controls (9.0 and 16.8%, respectively). Histologically, all these GCs with increased serum progastrin and amidated gastrins were of intestinal type and showed CagA and Hp seropositivity. Serum IL-8 and gastric pH, above their cutoff points (pH >4.5), and serum PG-I level below its cutoff point (44.2 microg/l) were observed in a significantly higher number of GC patients as compared to controls.. (1) GC patients have higher Hp and CagA seroprevalence than matched controls, confirming that CagA-positive Hp infection is associated with higher risk of GC; (2) serum levels of amidated gastrins and their precursor, progastrin, as well as IL-8 are significantly higher, while serum PG-I levels are reduced in intestinal type GC compared to controls, and (3) determination of high serum progastrin, amidated gastrins and IL-8 combined with low serum PG-I may be useful biomarkers of GC.

Topics: Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Biomarkers, Tumor; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Gastric Acid; Gastrins; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Middle Aged; Pepsinogen A; Protein Precursors; Risk Factors; Seroepidemiologic Studies; Stomach Neoplasms

To determine the changes of IL-8 and IL-8 mRNA in gastric and duodenal mucosa of children with Hp infection, to study the effect of Helicobacter pylori (Hp) infection on the expression of IL-8 and IL-8 mRNA, and to evaluate its possible roles in the pathogenesis of gastric and duodenal mucosal inflammation in Hp related gastroduodenal diseases.. Biopsy specimens were taken from the antral and duodenal mucosa on endoscopy in patients with or without Hp infection, which was diagnosed by urease test and Warshing-Starry staining. The expression of IL-8 in gastric and duodenal mucosa was determined by ELISA, the expression of IL-8 mRNA was determined by using RT-PCR.. Inflammation of gastric antral mucosa was more severe in Hp-positive group than in Hp-negative group. Active inflammation often existed on the basis of chronic inflammation in Hp-positive mucosa, and duodenal mucosa had mild chronic inflammation in Hp-positive group. Of 17 children who were not infected with Hp, 4 had pathologically normal gastric mucosa and had mild chronic gastritis, one child had an active chronic gastritis. Nineteen children were infected with Hp and all had chronic gastritis with signs of active inflammation. Gastric and duodenal mucosal IL-8 and IL-8 mRNA were higher in HP infected than in non infected children (IL-8: in gastric mucosa 24.66 - 177.77 pg/mg, 2.94 - 12.98 pg/mg, t = 12.34, P < 0.01; in duodenal mucosa: 4.28 - 47.76 pg/mg, 2.04 - 9.52 pg/mg, t = 7.18, P < 0.01. IL-8 mRNA: in gastric mucosa 2.37 - 4.99, 0.05 - 0.44, t = 29.29, P < 0.01; in duodenal mucosa 1.22 - 1.87, 0.01 - 0.23, t = 37.20, P < 0.01). Children with active chronic gastritis had higher interleukin-8 levels and IL-8 mRNA expression than those with inactive gastritis (IL-8 in gastric mucosa: 12.98 - 177.77 pg/mg, 2.04 - 10.43 pg/mg, t = 10.66, P < 0.01; in duodenal mucosa: 5.28 - 47.76 pg/mg, 3.19 - 8.14 pg/mg, t = 6.52, P < 0.01. IL-8 mRNA in gastric mucosa: 0.51 - 4.99, 0.01 - 0.44, t = 18.62, P < 0.01; in duodenal mucosa: 0.23 - 1.87, 0.01 - 0.20, t = 19.10, P < 0.01).. Higher expression of IL-8 and IL-8 mRNA was seen in Hp-positive gastric and duodenal mucosa and in active gastritis. IL-8 may play an important role in the local gastric and duodenal mucosal inflammatory infiltration with a large number of neutrophils when there is Hp infection.

Topics: Adolescent; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Gastroscopy; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Male; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

The unique environment of the human stomach makes it difficult to establish representative in vitro models for Helicobacter pylori that mimic the natural infection. The in vitro explant culture (IVEC) technique is based on coculture of human gastric explants with H. pylori, where bacteria-host interaction is studied on the basis of interleukin (IL)-8 secretion of the explant tissue in response to infection. In this study, it was shown that H. pylori attachment to gastric epithelial tissue was required for induction of representative inflammatory responses, assessed here by IL-8 production. Furthermore, IL-8 production by the explant tissue in response to H. pylori infection demonstrated that the explants were adequately responsive. The IVEC technique for studies of the interplay between H. pylori and the human gastric mucosa during conditions of experimental infections in vitro could add knowledge to our understanding of the complex bacteria-host cross-talk in vivo.

Topics: Adult; Biopsy; Colony Count, Microbial; Culture Techniques; Female; Fucosyltransferases; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Stomach Diseases

Disease-associated virulence factors of Helicobacter pylori may not be independent of one another. The aim was to determine which H. pylori virulence factor(s) was the most important predictor of severity of gastric inflammation or clinical outcome.. cag Pathogenicity island (PAI), vacA babA2, and iceA status were determined by polymerase chain reaction (PCR). oipA functionality was based on switch status determined by PCR-based sequencing. A backward stepwise multiple regression analysis was performed to determine which factor(s) was the most discriminating for clinical outcome as well as the relationship to mucosal histology (H. pylori density, neutrophil infiltration, intestinal metaplasia, and gastric atrophy) and mucosal interleukin 8 (IL-8) production.. H. pylori were obtained from 247 patients (86 with gastritis, 86 with duodenal ulcer, and 75 with gastric carcinoma). Although oipA status was closely linked to specific cag PAI, vacA, and babA2 genotypes, only oipA status remained in the final model to discriminate duodenal ulcer from gastritis (adjusted odds ratio [OR] = 5 and 95% confidence interval [CI] = 2.1-11.9). Among the factors, only a functional oipA was significantly associated with high H. pylori density, severe neutrophil infiltration, and high mucosal IL-8 levels (P < 0.001). oipA status had no relationship to gastric atrophic changes.. oipA functional status was related to clinical presentation, H. pylori density, and gastric inflammation. cag PAI, babA2, or vacA status appear important only as surrogate markers for a functional oipA gene.

Topics: Amino Acid Sequence; Bacterial Outer Membrane Proteins; Base Sequence; Female; Gastric Mucosa; Gastritis; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Pyloric Antrum; Virulence

Despite numerous epidemiological studies, the association between Helicobacter pylori infection and gastric cancer (GC) remains unexplained. This study was designed to determine the seropositivity of H. pylori and cytotoxin-associated gene A (CagA), serum gastrin and interleukin-8 (IL-8) levels as well as basal intragastric pH and maximal histamine-induced gastric acid outputs (MAO) in a large series of GC patients and controls.. 337 GC patients (118 men and 219 women; median age 59.4; range 21-87) and 337 controls randomized for sex and age entered the study. Serum IgG antibodies to H. pylori and CagA and serum levels of IL-8 were measured by enzyme-linked immunosorbent assay, while serum-amidated gastrin was determined by specific radioimmunoassay and correlated with gastric luminal pH.. The numbers of GC patients and controls involved in the study in various age groups, ranging from 20 to > 70 years, were similar, but overall H. pylori IgG seropositivity in GC patients was significantly higher (90.8%) than in controls (79.2%). The overall CagA seropositivity in GC patients was about double (58.2%) that in controls (25.2%). Serum gastrin levels over the calculated cut-off value (38.88 pM/L) were found in several-fold larger number in GC patients (48%) than in controls (8.3%) and. similarly, serum IL-8 values over the cut-off point (1.77 pg/mL) occurred in almost all (99.7%) GC patients but in only a few controls (0.3%). Basal intragastric pH above the cut-off point (pH = 4.50) was observed in about 58.2% of GC patients compared to 15.1% in controls, and strong correlation between the serum gastrin and gastric pH was found in GC but weak in controls. The cut-off value for MAO was 12.3 mml/h; MAO below this cut-off value occurred in 89.9% of GC patients and in only 4.7% of controls. A summary odds ratio (SOR) in GC for H. pylori IgG was 2.59 (95% Cl: 1.61-4.22) for CagA - 4.12 (95% Cl; 2.93-5.8), for serum gastrin - 10.25 (95%; 6.47-16.47) and for MAO - 15.2 (95% Cl; 9.45-39.82). Multivariable analysis of serum gastrin, IgG and CagA, and luminal pH and MAO values revealed that only gastrin and CagA have significant influence on GC formation (OR > 1 in logistic regression).. 1. CG patients show significantly higher H. pylori IgG and CagA seropositivity than dyspeptic age- and gender-matched controls, confirming that gastric infection with CagA expressing H. pylori greatly increases the risk of GC. 2. Serum gastrin levels in GC but not in controls are correlated with the rise in intragastric pH, indicating that excessive gastrin release in GC is affected by lower intragastric pH. 3. Serum gastrin level and CagA seropositivity are significantly increased in the majority of GC patients, and are the only variables in multivariable analysis to have a predominant influence on GC formation, which suggests that both these parameters may be implicated in H. pylori-related gastric carcinogenesis. 4. H. pylori-infected GC patients produce significantly more IL-8 than do non-GC controls, probably reflecting CagA-positive H. pylori-associated gastritis.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Bacterial; Antigens, Bacterial; Bacterial Proteins; Dyspepsia; Enzyme-Linked Immunosorbent Assay; Female; Gastric Acid; Gastrins; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin G; Interleukin-8; Male; Middle Aged; Radioimmunoassay; Stomach Neoplasms

Infection by Helicobacter pylori (Hp) has been linked to extradigestive pathologies including ischemic cerebral disease. The aim of our study was to assess the relationship between chronic Hp infection and ischemic stroke risk factors.. 80 patients (pts) aged 60-75 years with ischemic stroke confirmed by CT scans (group I) and 80 age- and gender-matched healthy controls (group II) were included into trial. Atherosclerotic plaques from 20 Hp positive pts were obtained at carotid endarterectomy for Hp DNA assessment by PCR. In all groups following parameters were determined; 1) the prevalence of Hp infection using (13)C-Urea Breath Test (UBT), 2) plasma anti-Hp and anti-CagA IgG and interleukin-8 (IL-8), and 3) plasma lipids and fibrinogen. Hp positive pts and controls received one-week anti-Hp therapy and after six months total cholesterol, low-density lipoprotein (LDL)-cholesterol, fibrinogen and IL-8 levels were re-examined.. Hp infection was detected by UBT in 83.75% of stroke pts but only in 65% of controls. CagA seropositivity was also significantly higher in stroke pts (57.5%) than in controls (33.75%). Plasma levels of cholesterol, LDL-cholesterol and fibrinogen as well as IL-8 were significantly higher in Hp positive subjects, especially in pts with ischemic stroke. Six months following successful anti-Hp therapy, the plasma levels of total cholesterol, LDL-cholesterol, fibrinogen and IL-8 were significantly lower than those in Hp positive stroke pts and controls.. Hp infection represents risk factor of ischemic stoke via an interaction of Hp cytotoxins or cytokines with atherosclerotic plaques in carotic arteries.

Topics: Aged; Antigens, Bacterial; Bacterial Proteins; Brain Ischemia; Breath Tests; Cholesterol; Cholesterol, LDL; Chronic Disease; Fibrinogen; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin G; Interleukin-8; Middle Aged; Risk Factors; RNA, Ribosomal, 16S; Stroke

To determine the role of host genetic factors in Helicobacter pylori infection, we examined the relation between gastroduodenal diseases and IL-1B polymorphisms in patients with H. pylori infection. In addition, we also compared gastric mucosal cytokine levels in those patients. We confirmed the findings that the IL-1B-31 C-to-T base transition was inverted in association with the -511 T-to-C base transition. There was no relation regarding to IL-1B polymorphisms and clinical outcomes. The gastric mucosal IL-1B level of the body of the stomach but not the antrum was significantly different among IL-1B genotypes. Furthermore, the IL-8 levels in the body were also higher in IL-1B-511C/C/ IL-1B-31TT than H. pylori negative patients. These findings suggested that IL-1B polymorphisms enhance not only IL-1-B production but also IL-8 production in the gastric body and may play an important role in the development of atrophic gastritis.

Topics: Cytokines; Gastric Mucosa; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Polymorphism, Genetic; Stomach Neoplasms

Associations of Helicobacter pylori genotypes with disease differ between Western countries and Asia. Therefore, we directly compared histopathological and in vitro responses to clinical isolates with similar genotypes. Sixty-three cagA(+) vacAs1/m1 H. pylori isolates (United States, n = 24; Japan, n = 39) and eight cagA-negative vacAs2/m2 strains were incubated with AGS cells, and supernatants were assayed for interleukin-8 (IL-8) and for DNA fragmentation. CagA tyrosine phosphorylation in AGS cells and the sequence of the putative HP0638 (oipA) signal sequence region were determined for 22 representative strains. HP0638 and/or cag island mutant strains were created and examined in IL-8 and CagA tyrosine phosphorylation assays. Levels of IL-8 induction and DNA fragmentation were similar in the U.S. and Japanese cagA(+) vacAs1/m1 isolates. All 10 of the isolates with the highest IL-8 induction and 8 of the 10 isolates with the lowest IL-8 induction had an in-frame oipA open reading frame, and all 10 of the isolates with the highest IL-8 induction and 7 of the 10 isolates with the lowest IL-8 induction induced CagA tyrosine phosphorylation in AGS cells. Eight isolates from gastric ulcer patients induced significantly more apoptosis in vitro, and more severe gastritis and atrophy in vivo, than other Japanese isolates. Disruption of HP0638 did not affect IL-8 induction or CagA tyrosine phosphorylation. Thus, H. pylori cagA(+) vacAs1/m1 isolates from the United States and Japan induce similar IL-8 and apoptosis levels. Inactivation of HP0638 does not alter epithelial responses mediated by the cag island in vitro. Assessment of apoptosis in vitro identified a group of H. pylori isolates that induce more severe gastric inflammation and atrophy.

Topics: Adult; Aged; Antigens, Bacterial; Apoptosis; Bacterial Proteins; DNA Fragmentation; Female; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Japan; Male; Middle Aged; Phosphorylation; United States

In recent studies, the involvement of mast cells in the pathogenesis of Helicobacter pylori infection was suggested. In the present study, using isolated canine gastric mucosal mast cells, we undertook to elucidate the effects of interleukin-8 (IL-8) and H. pylori on histamine release from these cells.. Enriched canine gastric mucosal mast cells (50% target cells) were incubated in Hanks medium with IL-8, or water extract or sonicate of H. pylori for 15 min at 37 degrees C. The content of histamine in the supernatants and the cell pellets after centrifugation was assayed with a histamine radioimmunoassay (RIA) kit.. IL-8 (50 ng/ml) and concanavalin A (20 microg/ml) significantly increased histamine release from enriched gastric mucosal mast cells. Dose-dependent stimulation of histamine release by IL-8 (5-50 ng/ml) was also seen. Water extract and sonicate of H. pylori (10(8) bacteria) increased histamine release from mast cells. A concentration-dependent stimulation of histamine release by water extract or sonicate was also seen. The maximal response of histamine release was seen at the highest concentration of the water extract or sonicate.. The results indicated that IL-8 and H. pylori had stimulatory effects on histamine release from canine gastric mucosal mast cells. The results imply that IL-8 and soluble factors of H. pylori may accelerate inflammation of the gastric mucosa via histamine release from mast cells.

Topics: Animals; Bacterial Proteins; Cells, Cultured; Concanavalin A; Dogs; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Histamine Release; In Vitro Techniques; Interleukin-8; Mast Cells; Stomach Diseases

NSAID use and Helicobacter pylori both cause damage to the gastric mucosa and can cause peptic ulcers. Our aim was to test the relationship between gastric mucosal polymorphonuclear leukocyte (PMN) infiltration and the severity of NSAID-induced gastric injury. H. pylori density, mucosal interleukin-8 (IL-8), and nitrite levels were assessed after receiving placebo and again after receiving 1000 mg of naproxen daily for three days. Histology was graded using a visual analog scale (0-5). IL-8 levels were assayed by ELISA and nitrite levels by Griess reaction. Eleven healthy volunteers with H. pylori infection entered. All had normal-appearing gastric mucosa after placebo. Postnaproxen gastric damage included three with none, one with mild, three with moderate, two with severe, and three were very severe mucosal injury (including one with an ulcer >5 mm). There was an inverse correlation between endoscopic score and the pH of the gastric juice post-therapy (R = -0.77, P = 0.004). There was no significant change in histologic or biochemical parameters from pretreatment levels. And none of the parameters (eg, PMN density) predicted endoscopic outcome. In conclusion, there was no relation between mucosal PMN density and endoscopic mucosa injury. PMN infiltration, while not predictive, may be a surrogate for an H. pylori infection-related increased risk of NSAID ulcers.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Cell Count; Female; Gastric Acidity Determination; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Interleukin-8; Male; Middle Aged; Naproxen; Neutrophils; Nitrites; Stomach Ulcer

Presence of the Helicobacter pylori adherence factor blood group Ag-binding adhesin (BabA; binding to Lewis(b) (Le(b))) is associated with ulcer disease, adenocarcinoma, and precancerous lesions. The importance of BabA for bacterial colonization and the inflammatory response is unknown. A total of 141 antral biopsies from H. pylori-infected patients were assessed in regard to the degree of granulocytic (G0 degrees--G3 degrees) and lymphocytic (L1 degrees--L3 degrees) infiltration. DNA genotypes of babA2 (the transcriptionally active gene of BabA), cagA, and vacAs1/2 were determined by PCR. Colonization density and Le(b) status on gastric epithelial cells were determined by immunohistochemistry. Real-time quantitative (TaqMan) RT-PCR determined mRNA expression of IL-8, TNF -alpha, and the Th1 markers IFN-gamma and the IL-12R beta2 chain. A total of 91% of infected patients were Le(b) positive. The vacAs1(+)/cagA(+) strains harboring babA2 showed significantly higher levels of granulocytic infiltration, bacterial colonization, and IL-8 mRNA than vacAs1(+)/cagA(+) strains lacking babA2. IL-8 mRNA and protein production by KATO III cells in vitro increased dose dependently with addition of different numbers of type 1 strains (G27 and 2808 strains, 0.1--20 bacteria/cell). The mRNA expression of TNF-alpha, IFN-gamma, and IL-12R beta2 was higher in H. pylori-positive patients than in controls, but it did not differ significantly between patients infected with different strain types. These data suggest that BabA facilitates colonization of H. pylori and thereby increases IL-8 response, resulting in enhanced mucosal inflammation. Infection with strains harboring BabA thereby augment a nonspecific immune response, whereas the Th1 response toward H. pylori appears to be independent of BabA, cytotoxin-associated gene A, or vacuolating cytotoxin.

Topics: Adhesins, Bacterial; Adjuvants, Immunologic; Adult; Aged; Aged, 80 and over; Atrophy; Carrier Proteins; Cell Line; Cell Movement; Colony Count, Microbial; Female; Gastric Mucosa; Genes, Bacterial; Genotype; Granulocytes; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Intestines; Lewis Blood Group Antigens; Male; Metaplasia; Middle Aged; Oligosaccharides; Pyloric Antrum; Th1 Cells

The sensitivity of Helicobacter pylori chromosomal DNA to MboI digestion was investigated in 208 strains from several continents. Only 11 (5%) of strains were sensitive to MboI, and it was hypothesized that HpyIII, a type II restriction/modification enzyme with sequence homology to MboI, mediated the protection. This was confirmed by PCR analysis of the gene locus of hpyIII, normally composed of hpyIIIR and hpyIIIM. In all but one strain sensitive to MboI, no PCR product of hpyIIIR was obtained. In contrast, all strains yielded a product for hpyIIIM, independent of MboI phenotype. Further examination of the hpyIII locus in strains lacking a hpyIIIR PCR product identified a novel gene, hrgA, upstream of hpyIIIM. All 208 strains examined had either hpyIIIR or hrgA, but not both, upstream of hpyIIIM. Although hrgA has homology with a Campylobacter jejuni gene (Cj1602), its function is not known. In Western countries, hrgA was more prevalent (53%) than in Asia (25%; P < 0.0001, chi(2)). In Asia, hrgA was more prevalent among gastric cancer patients (18 of 43; 42%) than among noncancer patients (16 of 95; 17%; P = 0.001, chi(2)). All 143 Asian strains tested were cagA(+), but among Western strains, hrgA was more prevalent in cagA(+) strains (26 of 42; 62%) than in cagA(-) strains (9 of 23; 39%; P = 0.04, chi(2)). In coculture with epithelial cells, hpyIIIR and hrgA strains did not show any significant differences in interleukin-8 induction and apoptosis. Although a direct function for hrgA in virulence could not be demonstrated, our data indicate that hrgA is a strain-specific gene that might be associated with gastric cancer among H. pylori isolates from Asian patients.

Topics: Adult; Aged; Bacterial Proteins; Deoxyribonucleases, Type II Site-Specific; DNA, Bacterial; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Stomach Neoplasms; Virulence

Adherence of Helicobacter pylori to the gastric epithelium is believed to be an important step in the induction of active inflammation of the mucosal layer. However, structural evidence showing a quantitative relationship between the adherence of H. pylori and severity of gastric mucosal inflammation is lacking. We therefore investigated the correlations between severity of gastritis and adherence of morphologically different forms of H. pylori. Fifty-seven biopsy specimens from the gastric bodies of patients with H. pylori-induced gastritis were examined. The severity of gastritis and the adherence and structure of H. pylori were determined with the use of light and scanning electron microscopy. We also investigated the ability of H. pylori organisms with different structural features to induce interleukin-8 secretion by human gastric adenocarcinoma (AGS) cells in vitro because production of interleukin-8 is related to H. pylori-associated gastritis. Furthermore, serum pepsinogen concentrations and cytotoxin-associated protein status in relation to adherence of H. pylori to the epithelial surface were examined. The results indicated that H. pylori organisms, which adhered firmly to the epithelial surface, were consistently long, tightly coiled bacilli. Histologically, those gastric mucosa samples with H. pylori firmly attached showed severe gastritis. H. pylori bacilli of greater length induced higher levels of interleukin-8 secretion. The serum pepsinogen I/II ratio showed a significant negative correlation with the grade of H. pylori adhesion (r = -0.401, P <.01). We also noted a significant correlation between cytotoxin-associated protein status and the adherence of H. pylori (r = 0.344, P <.05). A quantitative correlation was found between adherence of H. pylori and gastric inflammation. Both adherence and the induction of inflammation were found to be related to the structure of H. pylori.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Adhesion; Bacterial Proteins; Epithelial Cells; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Microscopy, Electron, Scanning; Middle Aged; Pepsinogen A; Severity of Illness Index; Virulence

Colonization of the gastric mucosa with Helicobacter pylori is associated with a dense infiltration of granulocytes into the lamina propria in the active phase of gastritis. In this study, we investigated the involvement of epithelial cell-derived neutrophil-activating protein 78 (ENA-78) in development of H. pylori-associated gastritis. Antral biopsies from 27 patients with H. pylori-associated gastritis and 25 from H. pylori-negative individuals were first analyzed for ENA-78 and interleukin-8 (IL-8) mRNA by semiquantitative reverse transcription (RT)-PCR. In H. pylori-positive patients, significantly elevated levels were found for both chemokines (P<0.05). Only IL-8 mRNA levels differed significantly (P<0.05) in H. pylori-infected individuals who had serum antibodies for cytotoxin-associated protein CagA versus H. pylori-infected CagA-negative persons. Quantification of ENA-78 transcript levels by competitive RT-PCR yielded a significant 45-fold upregulation for ENA-78 transcripts in biopsies of H. pylori-positive versus H. pylori-negative patients (P<0.05). In contrast to earlier findings with IL-8, the degree of ENA-78 mRNA upregulation was independent of the grade of activity of gastritis. Immunofluorescence studies on tissues of antral biopsies localized ENA-78 protein expression mainly to the gastric epithelium of H. pylori-positive patients, while control tissues were negative. Upregulation of ENA-78 and IL-8 mRNA and protein expression was also observed in an in vitro system using a gastric adenocarcinoma cell line. Only viable H. pylori yielded a strong ENA-78 and IL-8 induction, while H. pylori outer membrane proteins or water-soluble proteins had no significant effect. These data provide evidence for the importance of both IL-8 and ENA-78 in the development and perpetuation of H. pylori-associated gastritis.

Topics: Antigens, Bacterial; Bacterial Proteins; Chemokine CXCL5; Chemokines, CXC; Gastric Mucosa; Gastritis; Gene Expression Regulation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Helicobacter pylori virulence is associated with the presence of the cag pathogenicity island (PAI). The cag PAI is involved in the ability to induce interleukin-8 (IL-8) secretion by human cells, which is implicated in the inflammatory response of the gastric mucosa to H. pylori infection. The aim of this study was to determine whether the genetic structure of the cag PAI is conserved and whether it is linked to IL-8 induction ability. Detection of specific markers (cagA, picB, cag13-cag14, virD4, and IS605) by PCR and dot blot hybridization and long-distance PCR determination of the presence of cagI, cagII, and the middle region of the cag PAI were performed on 153 strains isolated from adults suffering from ulcers (n = 79) or gastritis (n = 74). IL-8 induction ability was evaluated by coculture of the strains with HEp-2 cells. Eighty-three strains (54.3%) had an entire cag PAI, 12 strains (7.8%) had the cag PAI split in two, 49 strains (32%) had no cag PAI, and 9 strains exhibited other structural combinations. The presence of an entire cag PAI was statistically correlated with the presence of IS605 (P = 0.006) and the ability to induce IL-8 secretion but not with clinical presentation of the infection. The structure of the cag PAI appears to be rather conserved and is related to the proinflammatory power of a strain. The existence of strains inducing IL-8 secretion regardless of the cag PAI structure suggests that this region is not the only requirement for IL-8 secretion.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Female; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Virulence Factors

The etiology and pathophysiology of stomach carcinoma is complex, and the mechanism whereby H. pylori directly or indirectly induces carcinoma remains unclear. In this study, interleukin (IL)-8, IL-4 and interferon (IFN)-gamma were measured in the tissue culture supernatant of gastric organ cultures from subjects with chronic gastritis with or without H. pylori infection, and with or without gastric cancer and gastric dysplasia.. Interleukin-8 levels were higher in cancer- and H. pylori-infected gastritis subjects than in H. pylori-negative subjects (12.95 +/- 3.16, 10.48 +/- 1.55 and 4.49 +/- 1.28 ng/mL, respectively). Elevated levels of IFN-gamma were detected in both H. pylori-infected and non-infected subjects with uncomplicated gastritis (72.23 +/- 19.0 and 34.61 +/- 5.30 pg/mL) and in non-infected dysplasia subjects (88 +/- 20.5 pg/mL). Background levels of IL-4 (< or = 9.4 pg/mL) in uncomplicated gastritis subjects and relatively high levels of IL-4 in dysplasia subjects (25.8 +/- 7.3 pg/mL) were detected. In contrast, trace amounts of IFN-gamma (16.01 +/- 0.35 pg/mL) and high levels of IL-4 (42.81 +/- 8.49 pg/mL) in gastric biopsy culture supernatants were found in cancer subjects. Mucosal IL-4 levels (but not IL-8 levels) correlated with infection and mucosal anti-H. pylori immunoglobulin G antibody.. The significant differences between gastritis with and without cancer and dysplasia indicated a shift from a Th1 to a Th2 helper cell pattern of cytokine secretion. This study has identified a local mucosal defect in gastric cancer. The near absence of IFN-gamma production from the mucosa at the margins of the tumor may be a critical factor in promoting growth of neoplastic cells.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antibodies, Bacterial; Female; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin A; Immunoglobulin G; Interferon-gamma; Interleukin-4; Interleukin-8; Male; Middle Aged; Stomach Neoplasms; T-Lymphocytes

Helicobacter pylori enhances the risk for ulcer disease and gastric cancer, yet only a minority of H. pylori-colonized individuals develop disease. We examined the ability of two H. pylori isolates to induce differential host responses in vivo or in vitro, and then used an H. pylori whole genome microarray to identify bacterial determinants related to pathogenesis. Gastric ulcer strain B128 induced more severe gastritis, proliferation, and apoptosis in gerbil mucosa than did duodenal ulcer strain G1.1, and gastric ulceration and atrophy occurred only in B128+ gerbils. In vitro, gerbil-passaged B128 derivatives significantly increased IL-8 secretion and apoptosis compared with G1.1 strains. DNA hybridization to the microarray identified several strain-specific differences in gene composition including a large deletion of the cag pathogenicity island in strain G1.1. Partial and complete disruption of the cag island in strain B128 attenuated induction of IL-8 in vitro and significantly decreased gastric inflammation in vivo. These results indicate that the ability of H. pylori to regulate epithelial cell responses related to inflammation depends on the presence of an intact cag pathogenicity island. Use of an H pylori whole genome microarray is an effective method to identify differences in gene content between H. pylori strains that induce distinct pathological outcomes in a rodent model of H. pylori infection.

Topics: Animals; Antigens, Bacterial; Apoptosis; Bacterial Proteins; Cell Division; Cell Line; Duodenal Ulcer; Gastric Mucosa; Gastritis; Genome, Bacterial; Gerbillinae; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Oligonucleotide Array Sequence Analysis; Sequence Deletion; Stomach Ulcer

The pathogenic role of human neutrophils has been implicated in Helicobacter pylori-associated gastritis. Ecabet sodium, a locally acting antiulcer drug, has anti-H. pylori actions. The aim of this study was to examine the effects of ecabet on the ability of H. pylori to stimulate human neutrophils. H. pylori were added to 1 x 10(5) neutrophils and incubated for 30 min in the presence of ecabet. Bacterial suspensions which had been incubated with ecabet for 30 min were also used to stimulate neutrophils. The intracellular production of reactive oxygen species was measured with a FACScan. Bacterial suspensions were also added to neutrophils in the presence of ecabet and incubated at 37 degrees C for 12 h to measure interleukin (IL)-8 production by enzyme-linked immunosorbent assay. The mean fluorescence intensity was found to be attenuated dose-dependently by ecabet (P < 0.01). Ecabet also inhibited IL-8 production by neutrophils in a dose-dependent manner (P < 0.001). Bacteria with prior incubation with ecabet induced significantly lower IL-8 production than those without this incubation (P < 0.05). Ecabet sodium has preventive effects on the ability of H. pylori to stimulate human neutrophils. It may lead to reduced gastritis activity and decreased oxidative damage of the gastric mucosa in H. pylori-associated gastritis.

Topics: Abietanes; Anti-Bacterial Agents; Anti-Ulcer Agents; Diterpenes; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Luminescent Measurements; Neutrophils; Reactive Oxygen Species

Interleukin 10 (IL-10) is a counter-inflammatory peptide implicated in the downregulation of human intestinal immune responses. Enhanced secretion of IL-10 has been documented in gastric biopsy organ culture in Helicobacter pylori infection. This study aimed to define the cellular origins of IL-10 in H pylori associated gastritis, and to determine the effects of endogenous IL-10 on proinflammatory cytokine secretion in vitro.. Endoscopic biopsies were obtained from the gastric antrum at endoscopy from patients with dyspepsia. Two pairs of antral biopsies were cultured in vitro for 24 hours, one pair in the presence of neutralising anti-IL-10 monoclonal antibody, the other pair as controls. The cytokine content of culture supernatants (tumour necrosis factor alpha (TNF-alpha), IL-6, and IL-8) was determined by enzyme linked immunosorbent assay and corrected for biopsy weight. Helicobacter pylori status was established by histology and biopsy urease test, and histopathology graded by the Sydney system. In a subgroup of patients, western blotting was used to establish CagA serological status. Immunohistochemistry for IL-10 was performed on formalin fixed tissues using a combination of microwave antigen retrieval and the indirect avidin-biotin technique. Immunoreactivity was scored semiquantitatively.. In vitro culture was performed in 41 patients: 31 with H pylori positive chronic gastritis and 10 H pylori negative. In vitro secretion of TNF-alpha, IL-6, and IL-8 for "control" biopsies was significantly higher in H pylori positive versus negative samples, with values of TNF-alpha and IL-6 correlating with the degree of active and chronic inflammation and being higher in CagA seropositive cases. No evidence for enhanced cytokine secretion was seen in biopsies cocultured in the presence of anti-IL-10 monoclonal antibody. Immunohistochemistry was performed in 29 patients, of whom 13 were H pylori positive. IL-10 immunoreactivity was observed in the surface epithelium in all H pylori positive cases and in 13 of 16 negative cases, especially in areas of surface epithelial degeneration. Lamina propria mononuclear cells (LPMNCs) were positively stained in all H pylori positive cases and in 12 of 16 negative cases, with a significantly greater proportion of positive LPMNCs in the positive group.. This study localised IL-10 protein to the gastric epithelium and LPMNCs. In vitro proinflammatory cytokine secretion was increased in H pylori infection (especially CagA positive infection), but blocking endogenous IL-10 secretion did not significantly increase cytokine secretion. IL-10 is implicated in H pylori infection and might "damp down" local inflammation. The role of gastric IL-10 secretion in determining the clinicopathological outcome of infection merits further study.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Case-Control Studies; Chronic Disease; Epithelium; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-10; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Organ Culture Techniques; Stomach; Tumor Necrosis Factor-alpha

Chemokines that play a primary role in active inflammation are increased in gastric mucosa infected with Helicobacter pylori. Cigarette smoking increases the risk of peptic ulcer disease and gastric cancer, whereas alcohol might exert an antibacterial role. The aim of this study was to examine the association between smoking or alcohol consumption and mucosal chemokine mRNA expression in H pylori associated gastritis.. Gastric biopsy specimens were obtained from 46 patients with dyspepsia who were infected with H pylori, and total RNA was extracted. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of three C-X-C chemokines (interleukin 8 (IL-8), growth related oncogene alpha (GRO alpha), epithelial neutrophil activating protein 78 (ENA-78)) and two C-C chemokines (regulated on activation normal T cell expressed and secreted (RANTES) and monocyte chemotactic protein 1 (MCP-1)).. GRO alpha and ENA-78 mRNA expression was significantly increased (p < 0.05) in 22 smokers compared with 24 non-smokers; however, no difference was seen in the expression of IL-8, RANTES, and MCP-1 mRNA. No differences were observed in chemokine mRNA expression in relation to alcohol consumption.. The increased C-X-C chemokine mRNA expression seen in smokers might play a role in inducing enhanced inflammatory activity in gastritis and the consequent severe diseases associated with H pylori infection.

Topics: Alcohol Drinking; Case-Control Studies; Chemokine CCL2; Chemokine CCL5; Chemokine CXCL1; Chemokine CXCL5; Chemokines; Chemokines, CXC; Chemotactic Factors; Chi-Square Distribution; Dyspepsia; Gastric Mucosa; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Smoking

The pathophysiology of hypergastrinemia in H. pylori infection has been largely investigated and different reports clearly show that the infected antrum has a marked inflammatory response with a suggestive local production of cytokines. Notwithstanding, a few data are available on the circulating levels of cytokines and gastrin in the asymptomatic people carrying H. pylori infection. Thus, aim of the study was to evaluate circulating proinflammatory cytokines [Interleukin (IL)-8, Interleukin (IL)-10, Interferon (IFN)-gamma, and Tumor Necrosis Factor (TNF)-alpha] and gastrin levels in H. pylori positive asymptomatic subjects vs. H. pylori negative ones. To this end, thirty healthy volunteers with no digestive symptoms or systemic disease were enrolled and H. pylori infection was identified by a 13C-urea breath test. Plasma levels of gastrin were determined using the RIA kit whereas IL-8, TNF-alpha, IL-10, and IFN-gamma levels in serum were measured with a solid-phase ELISA. Fifteen infected people showed significantly higher gastrin and TNF-alpha levels than uninfected subjects. On the contrary, IL-8 levels were significantly higher in the uninfected subjects than in H. pylori positive ones (P < 0.0422). IFN-gamma and IL-10 circulating levels were not affected by H. pylori presence, being not significantly different in the two groups.

Topics: Adult; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Gastrins; Helicobacter Infections; Helicobacter pylori; Humans; Interferon-gamma; Interleukin-10; Interleukin-8; Male; Middle Aged; Radioimmunoassay; Tumor Necrosis Factor-alpha

Further elucidation of the consequences of Helicobacter pylori infection on gastric mucosal inflammation and gastric secretory function would be facilitated by an animal model that is susceptible to infection with H. pylori, is broadly similar in gastric physiology and pathology to people, and is amenable to repeated non-invasive evaluation. The goal of this study was to examine the interrelationship of bacterial colonization, mucosal inflammation and gastric secretory function in cats with naturally acquired H. pylori infection.. Twenty clinically healthy cats with naturally acquired H. pylori infection (cagA-, picB) and 19 Helicobacter-free cats were evaluated. Gastric colonization was determined by tissue urease activity, light microscopy, culture and PCR. The mucosal inflammatory response was evaluated by light microscopy, and by RT-PCR of the pro-inflammatory cytokines IL-1alpha, IL-1beta, IL-8 and TNF-alpha in gastric mucosa. Gastric secretory function was assessed by measuring pentagastrin-stimulated acid secretion, fasting plasma gastrin, and antral mucosal gastrin and somatostatin immunoreactivity.. H. pylori colonized the pylorus, fundus and cardia in similar density. Bacteria were observed free in the lumen of gastric glands and were also tightly adherent to epithelial cells where they were associated with microvillus effacement. Mononuclear inflammation, lymphoid follicle hyperplasia, atrophy and fibrosis were observed primarily in H. pylori-infected cats, with the pylorus most severely affected. Neutrophilic and eosinophilic infiltrates, epithelial dysplasia, and up-regulation of mucosal IL-1beta and IL-8 were observed solely in infected cats. Fasting plasma gastrin concentrations and pentagastrin-stimulated acid output were similar in both infected and uninfected cats. There was no relationship of bacterial colonization density or gastric inflammation to plasma gastrin concentrations or gastric acid output.. The pattern of colonization and the mucosal inflammatory response in cats with naturally acquired H. pylori are broadly similar to those in infected people, particularly children, and non-human primates. The upregulation of IL-8 in infected cats was independent of cagA and picB. Our findings argue against a direct acid-suppressing effect of H. pylori on the gastric secretory-axis in chronically infected cats.

Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Cardia; Cat Diseases; Cats; Disease Models, Animal; Female; Gastric Acidity Determination; Gastric Fundus; Gastric Mucosa; Gastrins; Gastritis; Helicobacter Infections; Helicobacter pylori; Interleukin-1; Interleukin-8; Male; Pyloric Antrum; Reverse Transcriptase Polymerase Chain Reaction; Somatostatin; Tumor Necrosis Factor-alpha

In order to study the role of Helicobacter pylori infection in gastric carcinogenesis, we have measured oxidized (carbonyls) and nitrated (nitrotyrosine-containing) proteins as markers for oxidative and nitrative stress in 216 human gastric biopsies using dot and western immunoblots and correlated the results with H. pylori, cagA status, expression of interleukin-8 and inducible nitric oxide synthase (iNOS) mRNAs, and gastric pathology. Higher levels of both oxidized and nitrated proteins were found in patients with either chronic gastritis or duodenal ulcer than in those with normal mucosa. The levels of modified proteins were significantly higher in inflamed samples infected with H. pylori, especially cagA+ strains, and in those with expression of interleukin-8 and iNOS mRNAs than in those negative for these parameters. These results indicate that infection with cagA+ H. pylori induces significant oxidative and nitrative stress in stomach mucosa, contributing to the pathogenesis of H. pylori-associated gastroduodenal diseases.

Topics: Antigens, Bacterial; Bacterial Proteins; Biopsy; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrosation; Oxidative Stress; RNA, Messenger; Stomach

Helicobacter pylori infection stimulates several intracellular signaling pathways and is accompanied by increased gene expression in gastric epithelial cells. High-density cDNA microarray was used to characterize the mRNA expression profile of genes in human gastric cancer cells (MKN45, AGS) cocultured with H. pylori. Coculture with cag pathogenicity island (PAI)-positive H. pylori (wild-type) significantly up-regulated mRNA expression in 8 of 2304 genes tested. In 6 (interleukin-8, I(kappaB)alpha, A20, ERF-1, keratin K7, glutathione peroxidase) of the 8 genes, up-regulation was confirmed by RT-PCR. In coculture with isogenic cagE-negative mutant ((Delta)cagE), which encodes a type IV secretion system with other genes in the cag PAI, no significant up-regulation was found. We further analyzed the role of A20. Transfection of expression vector encoding A20 resulted in an inhibition of H. pylori-mediated NF-kappaB activation, indicating that H. pylori-mediated A20 expression could be a negative regulator of NF-kappaB activation. Taken together, these results indicate the importance of microarray technology as a tool for analyzing the complex interplay between H. pylori and the host.

Topics: Adenocarcinoma; Antigens, Bacterial; Bacterial Proteins; Coculture Techniques; DNA-Binding Proteins; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Helicobacter Infections; Helicobacter pylori; Humans; I-kappa B Proteins; Interleukin-8; Intracellular Signaling Peptides and Proteins; NF-kappa B; NF-KappaB Inhibitor alpha; Nuclear Proteins; Oligonucleotide Array Sequence Analysis; Proteins; RNA, Messenger; Stomach Neoplasms; Transcription Factors; Tumor Cells, Cultured; Tumor Necrosis Factor alpha-Induced Protein 3

Helicobacter pylori is etiologically involved in the development of gastric cancer and infected gastric mucosa has been shown to possess elevated levels of cytokines [for example interleukin (IL)-1beta, IL-6 and IL-8]. Because specific cytokines have also been shown to enhance the development of certain cancers, we examined the relationship between the levels of cytokines, the type and stage of gastric cancers, and the H. pylori infection.. Cytokines were measured from gastric cancer tissues, adjacent normal appearing mucosa, and the serum in 66 patients with early or advanced gastric cancer and from controls using semiquantitative RT-PCR and ELISA.. IL-6 and IL-8 levels were more than 10-fold increased in cancer tissues as compared with normal gastric tissues. IL-8 levels in cancer tissues were more than 2-fold higher in advanced gastric cancer as compared with early gastric cancer irrespective of H. pylori status. IL-6 levels were significantly higher in early gastric cancer with active H. pylori infection as compared with early cancer without H. pylori infection (8.7 + 1.4 vs. 1.2 + 0.3 pg/mg protein, p <.001) and decreased significantly after the cure of H. pylori (11.1 + 2.9-8.2 + 2.3 pg/mg protein, p <.05).. IL-8 levels in gastric cancer tissue are largely independent of H. pylori infection. In contrast, tissue IL-6 levels were high in H. pylori infected early gastric cancer and fell significantly after the cure of H. pylori suggesting a relationship between H. pylori infection and early gastric cancer.

Topics: Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; Stomach Neoplasms

To investigate whether Helicobacter pylori infection, but not drugs, affects gastric somatostatin, interleukin-8 (IL-8), histological inflammation through eradication therapy, and interactions among these parameters.. Twenty-eight H. pylori-positive patients (21 males; mean age 47.0 years) with either gastric ulcer (GU: n = 11) or duodenal ulcer (n = 17) diagnosed endoscopically were treated with dual therapy. Eradication was defined as negative microbiologic tests and 13C-urea breath test. Levels of antral and gastric juice somatostatin and mucosal IL-8 were measured by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. Histology was assessed by the Sydney system.. H. pylori was eradicated in 15 patients (10 males, 6 GU) out of 28 (54%). The patients' backgrounds did not affect the eradication of H. pylori. Successes in eradication significantly increased antral and juice somatostatin contents, and dramatically decreased IL-8 levels and histological gastritis. In contrast, persistent H. pylori infection did not affect somatostatin and histological gastritis. An inverse correlation was present between changes in somatostatin levels and histological activity. No relationship was observed in changed values between antral somatostatin and IL-8.. These results indicate that eradication of H. pylori, but not the drugs used, induced an increase in somatostatin levels in the antrum and gastric juice, suggesting a close relationship between H. pylori and gastric somatostatin regulation. A close correlation between an increase in gastric somatostatin levels and the normalization of histological activity was present, suggesting that certain peptide-immune interactions in the gastric mucosa exist in H. pylori infection.

Topics: Adult; Aged; Duodenal Ulcer; Endoscopy, Gastrointestinal; Female; Gastric Juice; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Peptic Ulcer; Pyloric Antrum; Somatostatin; Stomach; Stomach Ulcer

Helicobacter pylori infection might activate nuclear factor-kappaB (NF-kappaB), a transcriptional regulator of inducible expression of inflammatory genes, interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2). We studied the role of NF-kappaB on expression of IL-8 and COX-2 in H. pylori-stimulated AGS gastric epithelial cells by using antisense oligonucleotide (AS ODN) for NF-kappaB subunit p50 and an antioxidant, glutathione (GSH) as well as a NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC).. AGS cells were treated with p50 AS ODN, GSH or PDTC in the presence of H. pylori. mRNA expression and protein levels for IL-8 and COX-2 were determined by Northern blot analysis and Western blot analysis. Levels of IL-8, 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) and thromboxane B2 (TXB2) were measured in the medium by enzyme-linked immunosorbent assay. NF-kappaB activation was examined by electrophoretic mobility shift assay.. H. pylori induced a time-dependent expression of mRNA and protein for IL-8 and COX-2 via activation of NF-kappaB and increased the levels of IL-8, 6-keto-PGF1alpha and TXB2, which were inhibited by GSH and PDTC. H. pylori-induced expression of IL-8 and COX-2 was blocked in AGS cells transfected with p50 AS ODN.. NF-kappaB may play a novel role in expression of IL-8 and COX-2 in H. pylori-induced gastric inflammation.

Topics: Adenocarcinoma; Antioxidants; Cyclooxygenase 2; Drug Evaluation, Preclinical; Epithelium; Gastric Mucosa; Gastritis; Gene Expression Regulation, Neoplastic; Glutathione; Helicobacter Infections; Helicobacter pylori; Humans; Immunity, Mucosal; Interleukin-8; Isoenzymes; Membrane Proteins; NF-kappa B; Oligonucleotides, Antisense; Prostaglandin-Endoperoxide Synthases; Pyrrolidines; Stomach Neoplasms; Thiocarbamates; Time Factors; Transcription, Genetic; Tumor Cells, Cultured

Helicobacter pylori adheres to gastric epithelial cells and stimulates interleukin-8 production. Ceramide, a lipid second messenger, has become known as an important mediator of some actions of several cytokines. We have recently reported that H. pylori-dependent ceramide production may activate nuclear factor-kappaB and mediate interleukin-8 expression in human gastric cancer cell lines. In this study, we evaluated the effect of rebamipide, an antigastritis and antiulcer agent, on H. pylori-dependent ceramide production and subsequent interleukin-8 expression in Kato III cells. Rebamipide inhibited ceramide-induced interleukin-8 expression in a dose-dependent manner. Rebamipide decreased the ceramide-induced increase of the interleukin-8 mRNA level as assessed by Northern blotting. Rebamipide suppressed interleukin-8 gene transcription and nuclear factor-kappaB-dependent transcriptional activity as assessed by luciferase assay. Rebamipide inhibited the ceramide-induced degradation of IkappaB-alpha (a major cytoplasmic inhibitor of nuclear factor-kappaB), further supporting that rebamipide inhibits the activation of nuclear factor-kappaB. Rebamipide also inhibited the ceramide-dependent activation of mitogen-activated protein kinases. Furthermore, rebamipide significantly attenuated the H. pylori-dependent increase in the intracellular ceramide level. These results suggest a novel mechanism by which rebamipide may protect against the mucosal inflammation associated with H. pylori infection.

Topics: Alanine; Anti-Ulcer Agents; Blotting, Northern; Blotting, Western; Ceramides; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gastric Mucosa; Gene Expression Regulation, Neoplastic; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Mitogen-Activated Protein Kinases; NF-kappa B; Quinolones; Signal Transduction; Stomach Neoplasms; Tumor Cells, Cultured

Helicobacter pylori (HP) infection is usually accompanied by an increased plasma level of gastrin, a potent mitogen able to induce cyclooxygenase (COX)-2. This study examined (a) the seroprevalence of HP, its cytotoxic protein, CagA, and cytokines (tumor necrosis factor alpha, interleukins 1beta and 8) in 80 patients with colorectal cancers, before and after the removal of tumor, compared with 160 age- and gender-matched controls; (b) the gene expression of gastrin and its receptors (CCKB-R) in the cancer tissue, (c) the plasma levels and tumor tissue contents of gastrin, and (d) the mRNA expression of COX-1, COX-2, and apoptotic proteins (Bax and Bcl2) in cancer tissue and intact colonic mucosa. Anti-HP IgG, anti-CagA IgG seroprevalence, and cytokine levels were analyzed by enzyme-linked immunosorbent assay tests; gene expressions of gastrin, CCKB-R, COX-1, COX-2, Bax, and Bcl2 by reverse transcriptase polymerase chain reaction; and gastrin by radioimmunoassay. The seroprevalence of HP, especially that expressing CagA, was significantly higher in cancer patients than in controls and did not change 1 week after tumor resection while plasma cytokines were significantly reduced after this operation. Both gastrin and CCKB-R mRNA were detected in the cancer tissue and the resection margin; similarly, COX-2 mRNA was expressed in most of cancers and their resection margin but not in intact colonic mucosa, where only COX-1 was detected. The colorectal cancer tissue contained several folds more immunoreactive gastrin than cancer resection margin and many folds more than the intact colonic mucosa. We conclude that colon adenocarcinoma and its resection margin overexpress gastrin, its receptors, CCKB-R, and COX-2, and that HP infection may contribute to colonic cancerogenesis via overexpression of gastrin and COX-2, which may account for the stimulation of the tumor growth and the reduction in apoptosis as documented by enhanced mRNA expression of anti-apoptotic Bcl2 over proapoptotic Bax proteins.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Antigens, Bacterial; Apoptosis; Bacterial Proteins; Colorectal Neoplasms; Cyclooxygenase 2; Cytokines; Female; Gastrins; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Isoenzymes; Male; Membrane Proteins; Middle Aged; Prostaglandin-Endoperoxide Synthases; Tumor Necrosis Factor-alpha

Helicobacter pyloriinfection of the gastric mucosa in humans is usually acquired early in life. The chronic inflammation that ensues involves the increased production of inflammatory cytokines. Published data on production of these mediators by gastric mucosa of H. pylori-infected children are few.. Seventy-nine children, aged 5 to 18 years, referred for upper gastrointestinal endoscopy to four separate hospitals in Chile, were studied. The concentrations of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor alpha were measured in homogenates of gastric mucosal biopsy specimens. Cytokine expression was confirmed by reverse transcription polymerase chain reaction. These data were correlated with the patients' clinical, histologic and sociodemographic status.. Patient rate of colonization by H. pylori was inversely correlated with socioeconomic status (P < 0.005) and positively correlated with age (P < 0.0025). In gastric mucosa, concentrations of IL-1beta, IL-8, and tumor necrosis factor alpha were all significantly higher in H. pylori-positive patients than in H. pylori-negative patients and in patients who had histologic gastritis than in those with normal gastric mucosa. In patients with peptic ulcer disease, only IL-1beta and IL-8 concentrations were significantly elevated when compared with those of patients without ulcers. Interleukin-6 concentrations were comparable among the different groups analyzed.. This study suggests that increased gastric mucosal production of the proinflammatory cytokines IL-1beta and IL-8 is probably involved in H. pylori-associated gastric damage in children and may be crucial in determining the different clinical outcomes.

Topics: Adolescent; Age Factors; Biopsy; Child; Child, Preschool; Endoscopy; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Male; Peptic Ulcer; Reverse Transcriptase Polymerase Chain Reaction; Social Class; Tumor Necrosis Factor-alpha

The cytokine response during the acute phase of Helicobacter pylori infection in humans has not been studied. The aim of this study was therefore to investigate the early cytokine responses against H. pylori using cultured human stomach explants as a model of acute infection.. Gastric corpus tissue obtained from 13 adult uninfected and 3 H. pylori-infected patients undergoing gastric surgery due to obesity was used for preparation of mucosal explants. The cultured explants were exposed to different H. pylori strains or antigens, that is, lipopolysaccharides (LPS), urease and heat-shock protein (Hsp) B. The responses of the CXC chemokines interleukin (IL)-8, growth-related oncogene alpha (GROalpha) and interferon-inducible protein (IP) 10 as well as the CC chemokine regulated on activation normal T-cell expressed and secreted (RANTES) were determined by ELISA. In addition, IL-4, IL-6, IL-10, IL-12, interferon gamma (IFNgamma), tumour necrosis factor alpha (TNFalpha) and granulocyte-macropage-colony stimulating factor (GM-CSF) were studied.. In vitro H. pylori infection of the explants preferentially induced responses of the CXC chemokines GROalpha (P < 0.05) and IL-8 (P < 0.05), whereas the CC chemokine response (RANTES) was weak. In addition, the production of IL-6 was increased after H. pylori infection. Stimulation of the explants with different LPS preparations also induced strong GROalpha, IL-8 and IL-6 responses; the GROalpha responses being significantly higher after stimulation with rough than smooth H. pylori LPS (P < 0.05).. GROalpha, IL-8 and IL-6 are increased early during acute H. pylori infection and may influence the development of gastric disease.

Topics: Adult; Biopsy; Chemokine CCL5; Chemokines; Culture Techniques; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Granulocyte-Macrophage Colony-Stimulating Factor; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-6; Interleukin-8; Interleukins; Lipopolysaccharides; Male; Middle Aged; Receptors, Cytokine

The cagA-positive Helicobacter pylori strains are thought to be able to induce interleukin-8 expression and to be associated with gastroduodenal diseases. Inducible nitric oxide synthase (iNOS) may be involved in inflammatory pathogenesis. Our aim was to investigate the interrelationships between cagA and the expression of interleukin-8 and iNOS messenger RNAs, and with the type and degree of inflammation in gastric mucosa. In biopsies from 108 Chinese patients, the cagA, 16S rRNA, interleukin-8, and iNOS mRNAs were analyzed using reverse-transcription polymerase chain reaction. Specimens infected with cagA-positive strains had significantly more severe infiltration by mononuclear and polymorphonuclear leukocytes and more frequently expressed interleukin-8 and iNOS mRNAs than those infected with cagA-negative strains. iNOS and interleukin-8 mRNAs were significantly more frequently expressed together in the specimens with moderate or severe inflammation than in those with normal mucosa or mild inflammation. Our data suggest that interleukin-8 and excess nitric oxide play important roles in the pathogenesis of H. pylori-associated gastroduodenal diseases.

Topics: Antigens, Bacterial; Bacterial Proteins; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Prevalence; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Topics: Animals; Cytokines; Gastric Acid; Guinea Pigs; Helicobacter Infections; Helicobacter pylori; Interleukin-1; Interleukin-8; Parietal Cells, Gastric; Tumor Necrosis Factor-alpha

This study was undertaken to determine whether infection with Helicobacter pylori strains that contain the cagE gene was associated with duodenal ulceration in children. The presence of flaA, cagA, and cagE genes was determined by polymerase chain reaction in H. pylori previously cultured from 29 children. Twelve (92%) of 13 children with duodenal ulcers were infected with cagE-positive isolates, compared with only 5 (31%) of 16 with gastritis alone (P<.01). Infection of gastric cells in tissue culture by cagE-positive H. pylori resulted in greater increments in interleukin-8 levels compared with cagE-negative strains (2.3+/-0.1 vs. 1.3+/-0.2 ng/mL in AGS cells [P<.005]; 1.5+/-0.3 vs. 0.5+/-0.2 ng/mL in KATO-III cells [P<.05]). H. pylori-containing cagE was associated with the presence of duodenal ulceration in children. Enhanced chemokine production after infection with cagE-positive H. pylori could affect disease outcome.

Topics: Adolescent; Antigens, Bacterial; Bacterial Proteins; Cells, Cultured; Child; DNA, Bacterial; Duodenal Ulcer; Epithelial Cells; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Polymerase Chain Reaction; Retrospective Studies; Stomach

The transcription factor nuclear factor-kappaB (NF-kappaB) plays a pivotal role in inflammatory responses by upregulating mRNA expression of, for example, proinflammatory cytokines and chemokines. Although in vitro studies have shown that Helicobacter pylori can induce NF-kappaB activation in gastric cancer cell lines, there is little information on cellular localization of NF-kappaB in H. pylori-infected gastric mucosa.. H. pylori-infected and -negative patients with various endoscopic findings were examined. NF-kappaB expression was studied by means of Southwestern histochemistry, a new method of localizing transcription factors. Labeled double-stranded oligo-DNA with specific consensus sequence for the NF-kappaB binding site was reacted with frozen sections from gastric biopsy specimens. We compared mucosal interleukin-8 (IL-8) and IL-1beta levels as measured with enzyme-linked immunosorbent assay with the degree of NF-kappaB expression.. NF-kappaB expression was often evident in nuclei of epithelial cells in H. pylori-infected gastric mucosa. The degree of NF-kappaB expression on the epithelium was significantly more severe in H. pylori-infected than in -negative mucosa. The degree of NF-kappaB expression also correlated with mucosal IL-8 levels but not with IL-8.. H. pylori infection increases the expression of NF-kappaB in gastric mucosa, suggesting that NF-kappaB is involved in inflammatory responses to H. pylori.

Topics: Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Male; Middle Aged; NF-kappa B; Oligonucleotide Probes; Peptic Ulcer

Recent studies have shown that acid-suppressive therapy increases the severity of Helicobacter pylori- associated gastritis in the corpus.. To evaluate interleukin (IL)-8 production in the gastric corpus mucosa before and during acid-suppressive therapy in H pylori-infected patients.. Ten patients with reflux esophagitis (five H pylori-positive and five H pylori-negative) were treated with omeprazole 20 mg. Serum gastrin concentrations, H pylori colonization density and mucosal IL-8 levels in the corpus were investigated at entry and two weeks after starting therapy. IL-8 levels were measured by ELISA. The organism density was determined, and scored according to area occupied by the bacterial colonies. The presence of H pylori was assessed by rapid urease testing and histological finding of gastric biopsy specimens.. In H pylori-positive patients, concentrations of IL-8 during therapy significantly exceeded those before therapy (36.2+/-6. 8 versus 18.3+/-3.8 pg/mg protein; P<0.05) without altering H pylori density. In H pylori-negative patients, IL-8 levels were similar before and during therapy (6.1+/-2.7 versus 6.3+/-3.0 pg/mg protein). Concentrations of gastrin during therapy were significantly higher than those before therapy in all patients.. The results suggest that acid suppression increases mucosal IL-8 levels in H pylori-infected patients with reflux esophagitis.

Topics: Adult; Aged; Enzyme Inhibitors; Esophagitis, Peptic; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Omeprazole

Sofalcone has been reported to exert anti-ulcer and gastroprotective actions, but its exact mechanism of action remains unknown. In our laboratory, we found that indomethacin-induced gastric ulcers become worse when associated with Helicobacter pylori infection.. We employed the H. pylori-infected gnotobiotic murine model to examine the effect of sofalcone on indomethacin-induced gastric ulcers in the presence of H. pylori infection. In vitro experiments were also done to evaluate the effects of sofalcone on H. pylori growth, adherence of H. pylori to the MKN45 cells (a human gastric epithelial cell line), and these cells' IL-8 production in the presence of H. pylori.. We found that sofalcone produced a significant improvement in ulcer size as well as a substantial reduction in the number of H. pylori colonies in H. pylori-infected gnotobiotic mice. In vitro sofalcone has a significant bacteriocidal effect against H. pylori and can also significantly prevent adherence of this bacterium to MKN45 cells, thus remarkably reducing IL-8 production of these cells in response to stimulation by H. pylori.. Our results suggest that sofalcone can improve ulcer healing by the mechanisms mentioned above.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Anti-Ulcer Agents; Cell Adhesion; Chalcone; Chalcones; Disease Models, Animal; Helicobacter Infections; Helicobacter pylori; Humans; Indomethacin; Interleukin-8; Mice; Mice, Inbred BALB C; Stomach Ulcer

It has been suggested that gastric mucosal injury induced by Helicobacter pylori infection is mediated by interleukin-8 (IL-8).. We studied the effect of plaunotol, a drug extracted from the Plau-noi tree of Thailand, and reported it to be effective in the treatment of ulcers, of IL-8 secretion induced by H. pylori and of the inhibitory adhesion activity of the bacterium to gastric epithelial cells. Moreover, the therapeutic effect of plaunotol on H. pylori infection was assessed by using the gnotobiotic murine model.. Plaunotol inhibited the growth of H. pylori (1.5 x 10(4) c.f.u./mL) at high doses (24-48 microg/mL), but not at low doses (3-6 microg/mL). Interleukin-8 secretion induced by H. pylori was inhibited by coculture with plaunotol in a dose-dependent manner. The adhesion of H. pylori to MKN45 cells was also suppressed by coculture with plaunotol in a dose-dependent manner. An in vivo study showed that plaunotol improved histological gastritis and decreased the H. pylori antibody titre.. These findings suggest that plaunotol has a therapeutic effect on gastritis induced by H. pylori.

Topics: Administration, Oral; Animals; Anti-Ulcer Agents; Antibodies, Bacterial; Bacterial Adhesion; Diterpenes; Dose-Response Relationship, Drug; Epithelial Cells; Fatty Alcohols; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunoassay; Interleukin-8; Male; Mice; Mice, Inbred BALB C; Rabbits; Stomach Neoplasms; Stomach Ulcer; Tumor Cells, Cultured

The polymorphism of clinical presentations associated with Helicobacter pylori infection is potentially due to differences in the virulence of individual strains. H. pylori virulence has been associated with the ability to induce secretion of interleukin-8 (IL-8), the vacA genotypes, and the cagA status. The aim of this study was to determine the virulence profiles of 153 French H. pylori isolates on the basis of vacA genotypes, cagA status, and IL-8 induction ability. A total of 153 H. pylori isolates from patients with chronic gastritis (n = 74) or gastro-duodenal ulcers (n = 79) was examined for vacA genotypes and cagA status by polymerase chain reaction (PCR) and dot blot, and for their ability to induce IL-8 secretion by HEp-2 cells. The prevalence of vacA genotypes was: s1/m1 44.3%, s1/m2 24.9%, and s2/m2 23.5%. The cagA gene was present in 64% of the strains. IL-8 secretion was induced by 58.7% of the isolates. The presence of the cagA gene was significantly correlated with the s1/m1 vacA genotype and with the induction of IL-8. Thirty-four strains were atypical (cagA-positive/IL-8 noninducer or cagA-negative/IL-8 inducer). vacA genotypes, cagA status, and IL-8 induction ability are not correlated with the presence or absence of ulcer. The cagA status is not sufficient to predict the proinflammatory ability of H. pylori.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Alleles; Antigens, Bacterial; Bacterial Proteins; Female; Gastritis; Genes, Bacterial; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Immunoblotting; Interleukin-8; Male; Middle Aged; Peptic Ulcer; Polymerase Chain Reaction

Although increased levels of interleukin (IL)-8 are known to be associated with infiltration of neutrophils in the gastric mucosa with Helicobacter pylori infection, no study has investigated the relationship between local IL-8 levels and neutrophil infiltration in the duodenal mucosa of patients with duodenal ulcer (DU).. Duodenal mucosal biopsy specimens with and without gastric metaplasia (GM) were obtained from patients with DU and controls with an endoscopic methylene blue (MB) staining method. Levels of IL-8 secreted in the organ cultures of biopsy specimens were measured with an enzyme-linked immunosorbent assay. The number of myeloperoxidase-positive neutrophils infiltrating the lamina propria was determined in immunohistochemically stained tissue sections.. Histologic assessment showed that there was a strong correlation between the absence of endoscopic MB staining and the extent of GM. The levels of IL-8 in both duodenal and antral mucosal tissues were significantly higher in patients with H. pylori infection than in those without infection. In patients with DU the duodenal mucosal tissues with GM (MB-unstained mucosa) showed significantly higher levels of IL-8 than those without GM (MB-stained mucosa) or the antral mucosa. The number of neutrophils showed similar variations among DU and control patients with a positive correlation with IL-8 activity. The levels of IL-8 and the number of neutrophils decreased after H. pylori eradication in both duodenal and antral mucosal tissues, and these changes were more remarkable in the duodenal mucosal tissues with GM.. Increased IL-8 activity in the duodenal mucosa with GM may be important for ulcerogenesis in H. pylori-positive DU patients.

Topics: Adult; Aged; Biopsy; Duodenal Ulcer; Endoscopy, Digestive System; Enzyme-Linked Immunosorbent Assay; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Leukocyte Count; Male; Metaplasia; Middle Aged; Neutrophils; Stomach

Topics: Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Phenotype

Helicobacter pylori (Hp) infection is characterized by an intense inflammatory infiltrate in the gastric mucosa, which is chemoattracted by different cytokines. Interleukin-8 (IL-8) seems to play an important role in the recruitment of circulating neutrophils, and modulation of IL-8 secretion seems to be a strain marker. This study was designed to examine IL-8 concentrations in the gastric mucosa and their relationship with H. pylori phenotype and histologic findings.. Gastric biopsies were obtained from the antrum and corpus in 106 patients (69 Hp-positive and 37 Hp-negative). IL-8 levels in the gastric mucosa were analyzed by ELISA and Hp phenotype was determined with a western blot test.. 75% of H. pylori strains were CagA+ and 54.2% were VacA+. The Houston classification was used for histologic findings. No association between gastric atrophy or intestinal metaplasia and Hp phenotype was found. The highest IL-8 levels were found in CagA+ infected gastric mucosa, but the difference with respect to infection by a VacA+ strain was not statistically significant. IL-8 levels were highest when neutrophils were the predominant cell in the gastric inflammatory infiltrate (p < 0.05). IL-8 levels were higher in patients with atrophic gastritis than in patients with nonatrophic gastritis (p < 0.05).. In patients with H. pylori infection, IL-8 levels are higher than in Hp-negative patients regardless of Hp phenotype. There is an association between IL-8 and a neutrophilic infiltrate. Perpetuation of a chronic infiltrate could lead to more severe lesions such as atrophic gastritis or intestinal metaplasia, as deduced from the IL-8 levels found in these types of lesion.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Phenotype

Helicobacter pylori persists in the human stomach despite eliciting both cellular and humoral immune responses and inducing proinflammatory cytokines. To determine whether local humoral and cytokine responses are related to each other and to histologic responses, we studied 66 Japanese patients who underwent gastroscopy. Using specific enzyme-linked immunosorbent assays, we examined gastric antral mucosal-organ biopsy culture supernatants to assess interleukin-6 (IL-6) and interleukin-8 (IL-8) levels and antibody responses to H. pylori whole-cell antigens CagA, HspA, and HspB. Of the patients studied, 11 were H. pylori negative and 55 were H. pylori positive; by PCR, all strains were cagA(+). As expected, compared to H. pylori-negative patients, H. pylori-positive patients had significantly higher humoral responses to all H. pylori antigens and had higher IL-8 (47.8+/-3.5 versus 10.1+/-4.3 ng/mg of biopsy protein; P<0.001) and IL-6 levels (2.8+/-0.3 versus 0.26+/-0.2 ng/mg of protein; P<0.001). Among the H. pylori-positive patients, supernatant anti-CagA immunoglobulin G (IgG) levels were significantly associated with H. pylori density (P<0.005) and neutrophil infiltration (P<0.005) scores. Anti-CagA immunoglobulin A levels were correlated with intestinal metaplasia (P<0.05). Mononuclear cell infiltration scores were significantly associated with supernatant IL-6 levels (P<0.005) and with IgG responses to whole-cell antigens (P<0.05). Supernatant IL-8 levels were significantly associated with anti-CagA IgG (r = 0.75, P<0.001). Anti-CagA responses correlated with neutrophil infiltration, intestinal metaplasia, H. pylori density, and IL-8 levels, suggesting that the absolute levels of these antibodies may be markers for gastric inflammation and premalignant changes in individual hosts.

Topics: Antibodies, Bacterial; Antigens, Bacterial; Bacterial Proteins; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin G; Interleukin-6; Interleukin-8; Male; Organ Culture Techniques; Pyloric Antrum

Although mucosal alpha- and beta-chemokines are considered to be involved in the pathogenesis of Helicobacter pylori-associated gastritis, little is known how these chemokines are related to the ulcerogenesis in peptic ulcer patients. We examined the levels of interleukin (IL)-8 and macrophage inflammatory protein-1alpha (MIP-1alpha) in organ cultures and the numbers of inflammatory cells infiltrating the lamina propria by using the mucosal tissues obtained from gastric ulcer (GU) patients with and without H. pylori infection.. Levels of IL-8 and MIP-1alpha secreted in organ cultures were measured by an enzyme-linked immunosorbent assay. Numbers of myeloperoxidase-positive neutrophils, CD68-positive macrophages, and mononuclear cells were determined in tissue sections.. The mucosal tissues of both the gastric antrum and the ulcer site obtained from patients with H. pylori-positive GU showed significantly higher levels of IL-8 and MIP-1alpha and increased numbers of inflammatory cells compared with the corresponding mucosal tissues from those with H. pylori-negative GU or the antral mucosal tissues from H. pylori-negative controls. When the values were compared between the mucosal tissues from the gastric antrum and those from the ulcer site, the latter group of tissues showed significantly higher levels of IL-8 and MIP-1alpha and increased numbers of neutrophils and macrophages than the former group regardless of its healing process in patients with H. pylori-positive GU.. Mucosal alpha- and beta-chemokines may be important to the ulcerogenesis in H. pylori-associated GU disease.

Topics: Adult; Chemokine CCL3; Chemokine CCL4; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Macrophage Inflammatory Proteins; Male; Middle Aged; Stomach Ulcer

Helicobacter pylori (HP)-associated gastritis is characterized by an increased number of acute and chronic inflammatory cells secreting cytokines that contribute to maintain and expand the local inflammation. Locally induced IL-8 is believed to play a major role in the HP-associated acute inflammatory response. Factors/mechanisms that regulate IL-8 induction are, however, not fully understood. In the present study we investigated whether HP infection is associated with an increased production of IL-17, a T cell-derived cytokine capable of modulating IL-8 gene expression. We showed that both IL-17 RNA transcripts and protein were expressed at a higher level in the whole gastric mucosal and lamina propria mononuclear cell samples from HP-infected patients than in those from uninfected subjects. HP: eradication was associated with a marked down-regulation of IL-17 expression. The addition of a neutralizing anti-IL-17 Ab to the gastric lamina propria mononuclear cell cultures resulted in a significant inhibition of IL-8 secretion, indicating that IL-17 contributes to enhance IL-8 in the HP-colonized gastric mucosa. Consistently, stimulation of MKN 28 cells, a gastric epithelial cell line, with IL-17 increased IL-8 secretion. Finally, conditioned medium from the IL-17-stimulated MKN 28 cell cultures promoted the in vitro polymorphonuclear leukocyte migration. This effect was inhibitable by a neutralizing IL-8 but not IL-17 Ab. Together, these data indicate that biologically active IL-17 production is increased during HP: infection, suggesting the possibility that this cytokine may play an important role in the inflammatory response to the HP colonization.

Topics: Adjuvants, Immunologic; Adult; Aged; Cell Line; Cell-Free System; Cells, Cultured; Chemotaxis, Leukocyte; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; Neutrophils; Up-Regulation

Infection with Helicobacter pylori causes chronic active gastritis, which is characterized by neutrophils infiltrating the gastric epithelial layer and the underlying lamina propria as well as by T, B lymphocytes and macrophages accumulating in the lamina propria. In this study, the chemokine profile responsible for the recruitment of these inflammatory cells is investigated. Using both RNA/RNA in situ hybridization and immunohistochemistry, the expression of the neutrophil and/or lymphocyte-attractant CXC chemokines growth-related oncogene alpha (Gro(alpha)), IL-8, interferon-gamma (IFN-gamma)-inducible protein-10 (IP-10), monokine induced by IFN-gamma (MIG) and the CC chemokines macrophage inflammatory protein-1alpha (MIP-1alpha), -1beta, regulated on activation normal T cell expressed and secreted (RANTES) and monocyte chemoattractant protein-1 (MCP-1) is studied and microanatomically localized in the gastric mucosa. Macrophages in the lamina propria at sites with neutrophil infiltration and gastric epithelium infiltrated by neutrophils highly expressed the neutrophil-attractant chemokines Gro(alpha) and IL-8. Additionally, Gro(alpha) and IL-8 were expressed by neutrophils themselves localized within gastric epithelium, in the foveolar lumen and in the cellular debris overlying mucosal erosion. IP-10 and to a lower extent MIG, both selectively chemotactic for inflammatory T cells, were expressed by endothelial cells of gastric mucosal vessels and by mononuclear cells at sites with T cell infiltration. Expression of all other CC chemokines tested was significantly lower. These in vivo data indicate that a set of predominantly CXC chemokines modulates the inflammation in H. pylori gastritis. Gro(alpha) and IL-8 may play an important role in neutrophil trafficking from the mucosal vessel into the gastric epithelium, whereas IP-10 and MIG contribute to the recruitment of inflammatory T cells into the mucosa.

Topics: Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokine CXCL1; Chemokine CXCL10; Chemokine CXCL9; Chemokines, CXC; Chemotactic Factors; Gastric Mucosa; Gastritis; Gene Expression; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; In Situ Hybridization; Intercellular Signaling Peptides and Proteins; Interleukin-8; Macrophage Inflammatory Proteins; Neutrophils; RNA, Messenger; T-Lymphocytes

Helicobacter pylori infection is the commonest cause of gastritis. Different patterns of immune response to H. pylori infection and characteristics of bacteria are considered to contribute to clinical outcomes.. To determine characteristics of the host H. pylori relationship in subjects with non-ulcer dyspepsia and a histological diagnosis of gastritis.. Thirty-five subjects with chronic gastritis undergoing endoscopy (mean age 53 years, range 24-82, 14 male and 21 female) were studied, none of whom was on nonsteroidal anti-inflammatory drugs or antibiotics. H. pylori infection was determined by rapid urease test (CLOtest), culture, antibody and RT-PCR for Ure C, Cag A and 26 kDa gene and histology. Cytokine production of mucosal IL-6 and IL-8 were measured by ELISA.. Fifteen subjects were positive by CLOtest and/or bacterial culture. In these subjects histology showed numerous helical forms of H. pylori (Group I). Nine subjects were negative by CLOtest, bacterial culture, and mRNA for urease C fragment, but positive by PCR for the 26 kDa protein encoding gene. Histology in these subjects showed the presence of either coccoid forms (four), or scant helical forms (two), or mixed coccoid/helical forms (three) (Group II). Eleven subjects were negative by all methods of detection (Group III). IgG and IgA antibody levels in serum (p<0.05) and gastric tissue culture supernatant (p<0.001) were significantly higher in Group I than those in Group II or III. There were significant differences in the IgG serum and IgA supernatant antibody levels (p<0.01 and p<0.05) when Group II was compared to Group III. Supernatant IL-6 levels were significantly higher in Group I (p<0.01) than those from Groups II and III. IL-8 levels were higher in Group I (p<0.01) and Group II (p<0.05) when compared to Group III.. 'H. pylori-negative' gastritis can be associated with a non-urease producing form of H. pylori, with a reduction in both local and systemic antibody levels and mucosal pro-inflammatory cytokines.

Topics: Adult; Aged; Aged, 80 and over; Chronic Disease; Dyspepsia; Enzyme-Linked Immunosorbent Assay; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin A; Immunoglobulin G; Interleukin-6; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Urease

The effects of inflammatory cytokines induced by Helicobacter pylori infection on acid secretion have not been well defined. The purpose of this study was to investigate the direct effects of these cytokines on parietal cells isolated from guinea pigs. We examined the effects of human recombinant IL-1beta (0.05-100 ng/ml), IL-8 (2-256 ng/ml), and TNF-alpha (0.625-80 ng/ml) on acid secretion stimulated by three secretagogues (10(-4) M histamine, 10(-4) M carbachol, and 10(-5) M tetragastrin) and on basal acid secretion from isolated parietal cells, which was measured by the aminopyrine accumulation method. None of three cytokines showed any significant effects on stimulated or basal acid secretion from isolated guinea pig parietal cells. We concluded that inflammatory cytokines induced by Helicobacter pylori infection may affect acid secretion through mechanisms other than direct actions on parietal cells.

Topics: Aminopyrine; Animals; Cells, Cultured; Cytokines; Guinea Pigs; Helicobacter Infections; Helicobacter pylori; Interleukin-1; Interleukin-8; Male; Parietal Cells, Gastric; Tumor Necrosis Factor-alpha

Helicobacter pylori has been widely recognized as an important human pathogen responsible for chronic gastritis, peptic ulcers, gastric cancer, and mucosa-associated lymphoid tissue (MALT) lymphoma. Little is known about the natural history of this infection since patients are usually recognized as having the infection only after years or decades of chronic disease. Several animal models of H. pylori infection, including those with different species of rodents, nonhuman primates, and germ-free animals, have been developed. Here we describe a new animal model in which the clinical, pathological, microbiological, and immunological aspects of human acute and chronic infection are mimicked and which allows us to monitor these aspects of infection within the same individuals. Conventional Beagle dogs were infected orally with a mouse-adapted strain of H. pylori and monitored for up to 24 weeks. Acute infection caused vomiting and diarrhea. The acute phase was followed by polymorphonuclear cell infiltration, interleukin 8 induction, mononuclear cell recruitment, and the appearance of a specific antibody response against H. pylori. The chronic phase was characterized by gastritis, epithelial alterations, superficial erosions, and the appearance of the typical macroscopic follicles that in humans are considered possible precursors of MALT lymphoma. In conclusion, infection in this model mimics closely human infection and allows us to study those phases that cannot be studied in humans. This new model can be a unique tool for learning more about the disease and for developing strategies for treatment and prevention.

Topics: Acute Disease; Animals; Antibodies, Bacterial; Chronic Disease; Disease Models, Animal; Dogs; Endoscopy, Gastrointestinal; Female; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Male; Mice

Helicobacter pylori adheres to gastric epithelial cells, activates nuclear factor kappaB (NF-kappaB), and stimulates interleukin (IL)-8 production, but the responsible molecular mechanisms remain largely unknown. Because several studies have shown that sphingolipids are involved in a number of signaling pathways, including NF-kappaB activation, we investigated the possible role of sphingolipids in the regulation of IL-8 expression in Kato III and AGS cells.. IL-8 production in the conditioned media was quantified by enzyme immunoassay. Induction of messenger RNA (mRNA) was assessed by Northern blot analysis. Activation and binding activity of transcription factors were examined by luciferase assay and electrophoretic mobility shift assay, respectively. Intracellular levels of ceramide were quantified by diacylglycerol kinase assay.. A cell-permeable ceramide analogue (C2-ceramide) increased IL-8 expression with comparable mRNA induction. This effect was mimicked by sphingomyelinase, but not by phospholipase A2 or phospholipase C. C2-ceramide induced IL-8 gene transcription mainly through activation of NF-kappaB because mutation of the NF-kappaB-binding site completely abrogated the induction of luciferase activity. Direct contact of live H. pylori with epithelial cells increased the intracellular concentration of ceramide.. The results suggest a novel role of the sphingomyelin-ceramide pathway, at least in part through NF-kappaB, in IL-8 production induced by H. pylori infection in gastric epithelial cells.

Topics: Ceramides; Enzyme Inhibitors; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; NF-kappa B; RNA, Messenger; Sphingomyelin Phosphodiesterase; Sphingosine; Stomach Neoplasms; Transcription Factor AP-1; Transcription, Genetic; Tumor Cells, Cultured

Helicobacter pylori (H. pylori) appears to initiate an inflammatory cascade. Thus, phagocytes are accumulated in the gastric mucosa, in inflammatory conditions. Further, a potent chemotactic mediator, interleukin 8 (IL-8) is synthesized at such sites. The recently described IL-8 autoantibodies may, however, counteract the pro-inflammatory actions of IL-8. The aim was to study the correlation between H. pylori infection and IL-8, together with IL-8 autoantibodies in two different populations from a developed and a developing country.. Two different endoscopically characterized populations (65 Danes and 89 Albanians) were examined. IL-8 and IL-8 autoantibodies were detected by ELISA techniques, and H. pylori was identified by histological examinations.. Significantly more Albanian controls and dyspeptic patients (80 out of 89 persons) were H. pylori positive as compared to 24 of 65 Danes (p < 0.001). The median IL-8 level among Albanian controls 349 pg/mg protein was significantly higher than among Danes < 61 pg/mg protein (p < 0.001), and was at the same level as found in Danish peptic ulcer patients (p > 0.05). Further, H. pylori positive patients from both countries had significantly higher levels of IL-8 as compared to H. pylori negative patients (p < 0.001). However, significantly higher levels of IL-8 autoantibodies were found in the Albanian sub-population (median 138 O.D. units versus 52 O.D. units among Danes) (p < 0.001).. In H. pylori related disorders, a high mucosal IL-8 production has been found. However, this investigation further demonstrates higher levels of IL-8 autoantibodies among dyspeptic patients from a developing country, which might possibly counteract the pro-inflammatory actions of IL-8 by binding the molecule. The physiological significance of an altered immune response as described here needs to be elucidated in future studies.

Topics: Adolescent; Adult; Aged; Autoantibodies; Duodenal Ulcer; Duodenitis; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Stomach Ulcer

Increased production of proinflammatory cytokines, including tumour necrosis factor-alpha (TNF-alpha), IL-1beta, IL-6 and IL-8, has been demonstrated in Helicobacter pylori-associated gastric mucosal inflammation. IL-12, a newly characterized cytokine, is thought to be a key mediator in host responses to bacterial infections. The aim of this study was to investigate differences in cytokine patterns between H. pylori-positive and -negative gastritis and normal mucosa. Secretion of IL-12, TNF-alpha, IL-1beta, IL-6, IL-8 and IL-10 was measured in 176 patients with chronic gastritis in whole biopsy cultures. Gastritis was graded for chronic inflammation or acute inflammatory activity, respectively, according to the Sydney system. Biopsies with similar scores were matched for analysis from H. pylori-infected and non-infected patients. Secretion of IL-12 was significantly increased in H. pylori-associated gastritis in comparison with H. pylori-negative gastritis (P < 0.0001). In contrast, secretion of TNF-alpha, IL-1beta, IL-6, and IL-8 correlated with the degree of inflammation but was not different between H. pylori-positive and -negative patients. Moreover, IL-10 secretion was found to be higher in H. pylori-positive than in H. pylori-negative patients. IL-12 may play a specific role in H. pylori-associated gastric disease, whereas production of the proinflammatory cytokines TNF-alpha, IL-1beta, IL-6 and IL-8 does not seem to be restricted to H. pylori-induced inflammation. The contra-inflammatory cytokine IL-10 may be a contributor to the chronicity of H. pylori-associated gastritis by impairing clearance of the pathogen.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Chronic Disease; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

The present study aims at investigating the effects of mannitol and dimethylthiourea, known hydroxyl radical scavengers, on lipid peroxidation as an indicative of oxidative damage, NF-kappa B activation and IL-8 production by Helicobacter pylori in gastric epithelial cells. A human gastric epithelial cell line, AGS, treated with or without mannitol and dimethylthiourea, was incubated in the absence or the presence of H. pylori. As a result, H. pylori significantly stimulated the productions of lipid peroxide and IL-8. Treatment with H. pylori resulted in the activation of two species of NF-kappa B dimers (a p50/p65 heterodimer and a p50 homodimer). Mannitol and dimethylthiourea significantly inhibited lipid peroxide production, NF-kappa B complex formation and IL-8 production by H. pylori. In conclusion, mannitol and dimethylthiourea may attenuate H. pylori-induced gastric inflammation by inhibiting lipid peroxidation and NF-kappa B activation and thereby decreasing IL-8 production.

Topics: Cell Line; Cytokines; Diuretics, Osmotic; Electrophoresis; Epithelial Cells; Free Radical Scavengers; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Hydroxyl Radical; Interleukin-8; Lipid Peroxidation; Mannitol; NF-kappa B; Thiourea

We attempted to evaluate the relationship between the expression of IL-8 and RANTES and the dynamics of their target cells in human gastric mucosa of H. pylori associated gastritis, including their changes after H. pylori eradication. We performed the measurement of the mucosal level of IL-8 and RANTES protein by ELISA and immunohistochemistry. The neutrophil infiltration into the gastric mucosa was identified by the histological examination based on the Updated Sydney system and the measurement of MPO activity. The memory T lymphocyte and eosinophil were indicated by immunohistochemistry of CD45RO that is one of surface markers indicating memory T lymphocytes and MBP that is contained in the granules of eosinophils. H. pylori positive gastric mucosa demonstrated a remarkable increase in neutrophils. CD45RO positive cells and eosinophils, compared to H. pylori negative gastric mucosa. Gastric mucosal level of IL-8 and RANTES protein and MPO activity was significantly higher in H. pylori positive cases than that in H. pylori negative controls after H. pylori eradication, both of the level of IL-8 protein and MPO activity reduced at the same levels as negative controls. However, RANTES expression, CD45RO positive T lymphocytes and eosinophils remained in H. pylori eradicated gastric mucosa at the significantly high level, compared with H. pylori negative cases. Therefore, it seems possible that IL-8 might enhance the inflammation by facilitating the neutrophil infiltration into H. pylori infected gastric mucosa and that RANTES might play an important role in the specific immune response against H. pylori and the maintenance of the immune memory after H. pylori eradication.

Topics: Adult; Chemokine CCL5; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Immunologic Memory; Interleukin-8; Male; Neutrophils; T-Lymphocytes

Topics: Chronic Disease; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Peptic Ulcer; Reactive Oxygen Species

It is not known whether cagA+ Helicobacter pylori in duodenal ulcer (DU) have enhanced virulence compared with non-DU cagA+ H pylori.. To investigate the relation between presentation, H pylori density, interleukin 1beta (IL-1beta) and IL-8 production, and cagA status.. Fifty DU and 50 gastritis patients with cagA+ H pylori and 11 with cagA- infections were studied. Bacterial density and cytokine production were assessed using the same biopsies. Cytokine production was also measured from supernatants of medium following coculture of H pylori with MKN-45 cells.. There was no relation between H pylori density and cagA status. There was a dose dependent relation between mucosal cytokine levels and density of cagA+ H pylori. H pylori density increased to a threshold, followed by a rapid increase in cytokines and then a plateau. IL-1beta and IL-8 levels in the antrum were greater in DU than in gastritis; in the corpus the cytokine level/H pylori differed irrespective of similar H pylori densities. However, cytokine production was similar in vitro, independent of presentation or biopsy site, suggesting that host factors are critical determinants of the inflammatory response. Mucosal IL-8 and IL-1beta levels were low with cagA- and cagA+, cagE- H pylori infections.. The increase in antral IL-1beta and IL-8 production and inflammation in DU is related to increased numbers of bacteria and not to an increase in cytokine production per cagA+ isolate. There was no evidence of enhanced virulence of H pylori from DU compared with cagA+ non-DU H pylori.

Topics: Adult; Antigens, Bacterial; Bacterial Proteins; Biopsy; Duodenal Ulcer; Female; Gastritis; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Interleukins; Male; Middle Aged; Polymerase Chain Reaction; Tumor Cells, Cultured; Virulence

The monoclonal antibody (mAb) designated H20, which recognizes heat shock protein 60 (HSP60) of Helicobacter pylori, was previously established; and the epitope recognized by the mAb was shown to be species-specific. Using immunohistochemical staining of six gastric biopsy specimens with the H20 mAb, gastric epithelial cells of four biopsy samples stained positively. Flow cytometric analysis showed that H20 mAb reacted with primary human gastric epithelial (PHGE) cells, though the reactivities of the mAbs were different among the PHGE cells prepared. These results indicate that the species-specific epitope recognized by H20 mAb exists on human gastric cells. In addition, affinity-purified HSP60 from H. pylori by H20 mAb induced interleukin-8 (IL-8) secretion from PHGE cells (in one of four cases). These results indicate that H. pylori HSP60 induces IL-8 secretion from human gastric cells, and the levels of IL-8 differ among the various prepared PHGE cells.

Topics: Antibodies, Monoclonal; Antibody Specificity; Chaperonin 60; Epithelial Cells; Epitopes; Flow Cytometry; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Stomach Neoplasms; Tumor Cells, Cultured

The aim of this study was to examine the relation between disease specificity and the virulence factors of Helicobacter pylori isolated from patients with gastric cancer (GC), duodenal ulcer (DU), and gastritis (GS). Altogether 18 isolates obtained from patients with GC, 28 isolates from DU patients, and 13 isolates from GS patients were analyzed. All isolates were tested for the presence of the cagA gene, and genotyping of the vacA gene was done by the polymerase chain reaction. Production of VacA protein and expression of vacuolating cytotoxic activity in the H. pylori culture supernatant were examined. The serum antibody titers against purified VacA and CagA proteins were determined by enzyme-linked immunosorbent assay (ELISA). Interleukin-8 (IL-8) production by AGS cells in response to H. pylori isolates was measured by an hIL-8 ELISA kit. Genetic analysis of vacA revealed that most of the clinical isolates were classified into the S1a type by signal sequence typing. There were no differences in cagA detection rates, vacuolating cytotoxin activity, or mean antibody titers against VacA and CagA protein among the three groups. The mean IL-8 concentrations in the supernatants of AGS cells were similar in the three groups. In this study, there was no difference in virulence factors of H. pylori among isolates from GC, DU, and GS.

Topics: Antigens, Bacterial; Bacterial Proteins; Blotting, Western; Cytotoxins; Duodenal Ulcer; Enzyme-Linked Immunosorbent Assay; Female; Gastritis; Genotype; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin G; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Stomach Neoplasms; Virulence

It is well known that antrum-predominant gastritis and pan-gastritis occurs in the patients with Helicobacter pylori-positive duodenal ulcer (DU) and gastric ulcer (GU), respectively. However, the role of chemokines in the pathogenesis of these pathologies is unclear. We examined the regional differences in mucosal chemokine production in patients with DU and GU. The production of interleukin-8 (IL-8), growth-related gene (GRO) alpha, and macrophage inflammatory protein (MIP)-1alpha was greater in the antrum than in the corpus in DU patients. In the patients with GU, monocyte chemoattractant protein (MCP)-1 levels in the mucosa adjacent to ulcer were greater than those away for the ulcer in the corpus. The reduction in chemokine production occurring in association with the eradication of H. pylori differed between DU and GU patients in the antrum (IL-8, P = 0.0394; GROalpha, P = 0.0149; MIP-1alpha, P = 0.0246; MCP-1, P = 0.0087). The data imply a different pathogenesis may exist for the gastritis present in patients with DU and GU occurring in H. pylori-positive individuals.

Topics: Adult; Aged; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokine CXCL1; Chemokines; Chemokines, CXC; Chemotactic Factors; Duodenal Ulcer; Female; Gastric Mucosa; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Organ Culture Techniques; Pyloric Antrum; Stomach Ulcer

Strains of Helicobacter pylori carrying the virulence associated cag pathogenicity island (PAI) induce gastric epithelial synthesis of the chemokine interleukin-8 (IL-8), a neutrophil chemoattractant, and thereby a strong inflammatory response during chronic infection of the human gastric mucosa. Previous mutational analyses have shown that many genes in the cag PAI are needed to elicit IL-8 synthesis in gastric epithelial cells, and also that some genes are not involved.. To test the possibility that certain genes in the cag PAI also downregulate (modulate) the inflammatory response elicited by cag+ H pylori infection.. Cells of L5F11, a derivative of the Kato-3 gastric epithelial cell line that carries an engineered IL-8 promoter-luciferase reporter gene fusion, were cocultured with H pylori strain 26695 or with an isogenic mutant in which most of the cag PAI ORF 10 gene, an Agrobacterium virD4 homologue, was deleted. Luciferase activity was measured to assess IL-8 gene transcription and secreted IL-8 was measured by enzyme linked immunosorbent assay to assess synthesis and release of IL-8 protein from gastric epithelial cells.. Inactivation of ORF10 led to a 2.8-fold increase in IL-8 gene transcription and a 3.6-fold increase in IL-8 synthesis and secretion.. The results suggest that this VirD4 homologue participates in the control of inflammation that H pylori infection elicits by downregulating (modulating) the strong induction of IL-8 synthesis mediated by other cag encoded proteins.

Topics: Bacterial Proteins; Cell Line; DNA, Bacterial; Down-Regulation; Epithelial Cells; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Transcription, Genetic; Virulence; Virulence Factors

The role of Helicobacter pylori (HP) as the main etiological factor in gastritis and peptic ulcer disease is undisputable. Gastric mucosal damage caused by HP involves various bacterial and host-dependent toxic substances that have been recently associated with an increased risk of coronary artery disease (CAD), possibly through the activation acute phase response and of procoagulant hemostatic factors. Recent studies showed a close and strong correlation between plasma increments of some cytokines such as IL-6 or TNFalpha and cardiovascular diseases. HP infection induces platelet activation and aggregation that could be the pathogenic explanation of the association between HP infection and CAD. The aim of this study was to determine the seroprevalence of HP infection and antibodies to CagA, an antigen that is expressed by the most virulent HP strains inducing an enhanced gastric inflammatory response, in patients undergoing routine coronary artery examination. We studied 76 patients with CAD and 81 healthy controls patients without significant change in coronary circulation. Angiograms were read by two independent experienced cardiologists blinded to the results of HP status. The presence of serum IgG antibodies to HP and to CagA and plasma interleukin-8 (IL-8) levels was measured by ELISA. In addition plasma C-reactive protein fibrinogen, total cholesterol and lipids levels were measured in all studied patients. Seropositivity to HP was found in 81.5 % of cases and in 51% of controls and the difference in prevalence was statistically significant, the odds ratio being 4.3 for Hp patients. Antibody to CagA protein was detected in 47.3% of CAD but only in 28% of healthy controls (OR = 2.3 vs OR = 10). C-reactive protein, plasma fibrinogen and total cholesterol were, respectively higher in patients with CAD than in controls. Present data show that there is significant link between CAD and HP infection. The HP infection significantly increases the risk of CAD, especially when both the anti-HP IgG and anti-CagA IgG are considered. Higher prevalence of cytotoxic HP strains might enhance the atherosclerotic process by inducing a persistent, low grade inflammatory response in arterial wall with enhanced synthesis of acute phase reactants.

Topics: Adult; Aged; Antibodies, Bacterial; Antigens, Bacterial; Bacterial Proteins; C-Reactive Protein; Cholesterol; Coronary Disease; Female; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Triglycerides

To investigate the role of cytokines and antral gastritis in the development of duodenal ulcer that associated with Helicobacter pylori (H. pylori) infection.. 48 cases of duodenal ulcer with different degrees of H. pylori infection (35 cases with H. pylori infection and 13 patients without H. pylori infection,) were investigated by measuring the level of gastric mucosal cytokines and the degree of antral gastritis.. Compared with H. pylori negative cases, the patients with H. pylori positive duodenal ulcer had higher level of gastric mucosal IL-8, TNF alpha, MPO, MDA and severer antral gastritis. IL-8, TNF alpha, MPO and MDA were positirely correlated with H. pylori density and they were highly correlated with each other. However IL-6 did not have such specific properties.. Cytokines may be an important factor that links gastritis and duodenal ulcer associated with H. pylori infection.

Topics: Adult; Aged; Cytokines; Duodenal Ulcer; Female; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Pyloric Antrum; Tumor Necrosis Factor-alpha

C-X-C Chemokines play an important role for neutrophil extravasation through microvessels. Although the level of interleukin (IL)-8 is known to increase in the Helicobacter pylori-infected gastric mucosa, another C-X-C chemokine, GROalpha, has not been evaluated in the H. pylori-associated gastric mucosal injury. The present study was designed to investigate gastric contents of GROalpha in relation to those of IL-8 in the gastric mucosa of H. pylori-infected peptic ulcer patients. Thirty-eight patients with gastric ulcer and 41 with gastritis underwent endoscopy with informed consent and 49 were found to be H. pylori positive and 30 H. pylori negative. Biopsies from the gastric corpus were performed in each patient to examine the H. pylori colonization by bacterial culture, the rapid urease test and histological specimens as well as measurement of the contents of human GROalpha and IL-8. Helicobacter pylori infection was eradicated in 21 patients by triple therapy (lansoprazole 30 mg, amoxycillin 2.0 g, clarithromycin 600 mg; 2 weeks). The samples for GROalpha and IL-8 assay were homogenized in 0.02% aprotinin containing phosphate-buffered solution and the mucosal contents of GROalpha and IL-8 in the supernatants were quantified by sandwich enzyme immunoassay methods. The levels of GROalpha and IL-8 in H. pylori-positive gastric mucosa were significantly higher than those in the H. pylori-negative mucosa. There was a significant linear correlation between the levels of GROalpha and IL-8 (r = 0.798, P < 0.01). After the eradication of H. pylori by the triple therapy, the levels of GROalpha and IL-8 were significantly decreased. The GROalpha showed an increase in the H. pylori-positive gastric mucosa in a similar fashion as IL-8 contents, suggesting a pathogenetic role for GROalpha in H. pylori-associated gastric mucosal injury.

Topics: 2-Pyridinylmethylsulfinylbenzimidazoles; Adult; Aged; Anti-Bacterial Agents; Anti-Ulcer Agents; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Gastric Mucosa; Gastritis; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Lansoprazole; Middle Aged; Omeprazole; Stomach Diseases; Stomach Ulcer

Although chemokines have been suggested to play an important role in Helicobacter pylori associated gastritis, few studies have investigated the role of chemokines other than interleukin 8 (IL-8) in gastric mucosa.. To investigate the expression and production patterns of various chemokines using gastric biopsy specimens.. In 192 patients, expression patterns of C-X-C chemokines (IL-8 and growth regulated alpha (GRO alpha)) and C-C chemokines (regulated on activation, normal T cell expressed and presumably secreted (RANTES), monocyte chemotactic and activating factor (MCAF), macrophage inflammatory protein 1 alpha (MIP-1 alpha), and MIP-1 beta) were examined using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). cagA gene was identified using PCR.. H pylori infection was associated with increased rates of expression of mRNA for IL-8, GRO alpha, RANTES, and MIP-1 alpha and with increased levels of mucosal IL-8 and GRO alpha. IL-8 and GRO alpha levels correlated with the density of H pylori in both the antrum and corpus. The levels of these chemokines correlated with cellular infiltration in the antrum but not the corpus. cagA gene positive H pylori infection was associated with increased rates of expression of mRNA for IL-8 and GRO alpha and with increased levels of these chemokines.. H pylori infection is associated with increased expression rates and production of C-X-C chemokines (IL-8 and GRO alpha), but not with increased production of C-C chemokines. Although H pylori infection is associated with increased C-X-C chemokines in the antrum and corpus, there is a difference in the inflammatory response between these two areas of the stomach.

Topics: Adult; Aged; Aged, 80 and over; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Polymerase Chain Reaction; Pyloric Antrum; RNA, Messenger

Helicobacter pylori produces a gastric mucosal inflammation characterized by neutrophil infiltration, due to the liberation of interleukin-8.. To measure interleukin-8 levels in gastric mucosa samples from children colonized by H. pylori.. Thirty one children that required an upper gastrointestinal endoscopy for diagnostic purpose were studied. Antral biopsies were obtained for pathological study, H. pylori detection using CLO-test and interleukin-8 determination by ELISA.. Nine children were not infected with H. pylori. Of these, six had a pathologically normal gastric mucosa and three had a mild chronic gastritis. Twenty two children were infected by H. pylori and all had a chronic gastritis with activity signs in 13. Mucosal interleukin-8 was higher in infected than in non infected children (59.7 (range 6.1-379.7) and 15.8 (range 3.9-104.1) pg/mg respectively p = 0.029). Colonized children with an active chronic gastritis had higher interleukin-8 levels than those with inactive gastritis (84.4 (range 33.3-379.0) and 26.8 (range 6.1-372.6) pg/ml respectively p = 0.04).. Stomach colonization by H. pylori is associated with higher mucosal levels of interleukin-8. This phenomenon probably plays a role in the genesis and intensity of gastric mucosal inflammation in children.

Topics: Adolescent; Biopsy; Child; Child, Preschool; Chronic Disease; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male

Early studies suggested that two Helicobacter pylori proteins, CagA and VacA, were virulence factors. Support for that hypothesis has been undermined by geographic differences in prevalence of these antigens. To identify other possible putative virulence factors by establishing a relationship between antigens and different H. pylori diseases, two commercial available immunoblot assay kits, HelicoBlot 2.0 (Genelabs Diagnostics, Singapore) and RIDA Blot Helicobacter (R-Biopharm GmbH, Darmstadt, Germany), were used to investigate the prevalence of various specific antigen seropositivity in 80 H. pylori-infected Japanese (20 each with gastritis, duodenal ulcer, gastric ulcer, or gastric cancer). The production of interleukin-8 (IL-8) in biopsy specimens was also measured by enzyme-linked immunosorbent assay (ELISA). Both assays had 100% sensitivity; specificity was 90% for HB2.0 and 80% for RIDA-BH. With the exception of the 33-35 K antigen, there was no relationship between antigens, endoscopic diagnoses, histological findings, or mucosal IL-8 levels. The 33-35 K antigen was present in 97.5% (39 of 40) patients with gastric or duodenal ulcer compared to 70% (14 of 20) those with chronic gastritis (P < 0.006). The mean IL-8 levels in the corpus was significantly higher in those with antibody to the 33-35 K antigen compared to those without (105.4+/-22 pg/mg vs 10.2+/-8.8 pg/mg) (P=0.015). There was no relationship between other antigens including CagA and production of IL-8. In conclusion, the low-molecular-weight 33-35 K antigen may play an important role in the pathogenesis of H. pylori-related disease.

Topics: Antibodies, Bacterial; Antigens, Bacterial; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunoblotting; Interleukin-8; Japan; Male; Middle Aged; Molecular Weight; Prevalence; Reagent Kits, Diagnostic; Sensitivity and Specificity; Virulence

There is differential resolution of mucosal infiltration with neutrophils and mononuclear cells following successful Helicobacter pylori eradication. We investigated the effects of H. pylori eradication on mucosal interleukin-8 (IL-8) and IL-6 activity in relation to the resolution of H. pylori-associated gastritis. Eighty-one duodenal ulcer patients with H. pylori infection received dual- or triple-treatment eradication therapy, and mucosal biopsy specimens obtained at the initial and follow-up endoscopic examinations were cultured in vitro for 24 h. The levels of IL-8 and IL-6 were measured by enzyme-linked immunosorbent assays. In the 42 patients in whom H. pylori eradication failed, there was little change in the numbers of neutrophils and mononuclear cells infiltrating the mucosa and in IL-8 and IL-6 activity. In the 39 patients in whom H. pylori was eradicated, there was normalization both in the numbers of infiltrating neutrophils and in mucosal IL-8 activity, which was evident within 1 month following therapy. In contrast, there was a gradual resolution of mononuclear cell infiltration over a 6-month period, accompanied by a gradual normalization in IL-6 levels. Addition of H. pylori to cultures of mucosal tissues induced a significant increase in IL-8 activity in both uninfected control subjects and patients from whom H. pylori was eradicated. However, this introduction yielded a significant increase in IL-6 activity only in the latter group. This study indicates a dichotomy in the changes of mucosal IL-8 and IL-6 activity after H. pylori eradication. The rapid normalization of IL-8 after H. pylori eradication and the ability of H. pylori cells to stimulate IL-8 in control tissues indicate that IL-8 induction is a part of the innate (nonimmune) responses to this organism. In contrast, the results of experiments analyzing IL-6 activity in cultured mucosal tissues suggest that the gradual resolution of mucosal IL-6 activity and mononuclear infiltration after successful eradication observed in vivo may reflect gradually diminishing residual immune responses against H. pylori.

Topics: Adult; Anti-Bacterial Agents; Anti-Ulcer Agents; Cell Movement; Duodenal Ulcer; Female; Helicobacter Infections; Humans; Imidazoles; Interleukin-6; Interleukin-8; Intestinal Mucosa; Leukocytes, Mononuclear; Male; Middle Aged; Neutrophils; Organ Culture Techniques; Treatment Failure

To elucidate the role of interleukin (IL)-6 and IL-8 in the pathogenesis of gastric ulcer in Helicobacter pylori-positive gastritis, in situ hybridization using digoxigenin-labeled cDNA probes for both cytokines was performed. Immunogold silver staining was added to further improve the sensitivity of this non-radioactive hybridization. The biopsy specimens were taken from eight patients with active gastric ulcer before treatment, in all of whom H. pylori was positive. Macrophages (the putative producers of these cytokines) were determined by immunohistochemistry using anti-CD68 monoclonal antibodies (KP-1). IL-6 mRNA was most abundantly expressed in the epithelium and in the infiltrating cells in tissue adjacent to gastric ulcer. Quantitative analysis disclosed a significant increase in cells positive for IL-6 mRNA near the ulcer margin compared to cells in the surrounding tissue. In contrast, cells positive for IL-8 mRNA were observed in equal proportions and evenly in the epithelium and over the entire layer of the gastric mucosa regardless of the presence of gastric ulcer. The majority of infiltrating cells positive for both IL-6 and IL-8 mRNA were thought to be macrophages because of their morphologic features and their immunohistochemical reactivity to CD68. These findings strongly suggest that IL-6 is overexpressed at the margin of gastric ulcer in H. pylori-positive gastritis.

Topics: Adult; Aged; DNA Probes; DNA, Complementary; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; In Situ Hybridization; Interleukin-6; Interleukin-8; Male; Middle Aged; Peptic Ulcer; Polymerase Chain Reaction; RNA, Messenger; Up-Regulation

It is well known that Helicobacter pylori can cause gastritis, gastroduodenal ulcers and malignant diseases. The infiltration of polymorphonuclear leukocytes is recognized in the lesions of these diseases, and the infiltration disappears by antibiotic therapy. However, it is not yet clarified how Helicobacter pylori induces the formation of lesions including leukocyte infiltration. Recently, we have confirmed that several kinds of cytokines are expressed in the gastric biopsy specimens of gastroduodenal diseases. Especially, it is conjectured that chemokines such as interleukin-8 (IL-8) which are expressed in the specimens, induce leukocyte infiltration, gastric mucosal inflammation and gastroduodenal ulcers. It is possible that Helicobacter pylori CagA gene is closely related with IL-8 expression because this cytokine is more strongly expressed in the specimens from the patients infected with CagA-positive Helicobacter than those with CagA-negative one.

Topics: Anti-Bacterial Agents; Antigens, Bacterial; Bacterial Proteins; Cytokines; Duodenal Ulcer; Gastrointestinal Diseases; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Stomach Ulcer

We examined the relationship between the levels of macrophage inflammatory protein 1alpha (MIP-1alpha) and interleukin-8 (IL-8) in organ cultures of antral mucosal tissues, background gastroduodenal diseases, and grades of histologic gastritis. Significantly higher levels of MIP-1alpha and IL-8 were detected in patients with H. pylori infection than in those without infection. In H. pylori-positive patients, mucosal specimens from patients with peptic ulcer disease showed higher levels of MIP-1alpha and IL-8 than the specimens obtained from patients with erosive gastritis or those from endoscopically normal mucosa, and this was particularly pronounced in patients with duodenal ulcer. There were positive correlations between MIP-1alpha and IL-8 levels and histologic grades of activity, inflammation, and H. pylori density as defined by the Sydney system. However, the degree of association with the inflammatory cell count was different between these two chemokines. MIP-1alpha levels had a stronger association with mononuclear cells than with neutrophils, whereas IL-8 levels showed an association with neutrophils and mononuclear cells to an almost equal degree. These results suggest that MIP-1alpha and IL-8 may play important roles as inflammatory mediators in the pathogenesis of histologically proven H. pylori-associated gastritis.

Topics: Adult; Case-Control Studies; Chemokine CCL3; Chemokine CCL4; Culture Techniques; Duodenal Ulcer; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Pyloric Antrum; Stomach Ulcer

The development, analytical performance and applications of chemiluminescence imaging as a tool for quantitative analyte localization in target biological specimens are described. The detection of acetylcholinesterase activity both in array format and on a target surface are described. A proposed application of the method is a 384 well microtiter format assay for high throughput screening of acetylcholinesterase inhibitors such as tacrine, a drug widely used in the treatment of Alzheimer's disease, and two recently developed analogues. The chemiluminescent system in conjunction with optical microscopy allowed localization of acetylcholinesterase in brain tissue sections. We also describe the chemiluminescent immunohistochemical localization of interleukin 8 in Helicobacter pylori infected gastric mucosa cryosections and an in situ hybridization assay for the detection of herpes simplex virus DNA in single cells.

Topics: Acetylcholinesterase; Animals; Biological Assay; Brain; DNA, Viral; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Herpesviridae Infections; In Situ Hybridization; Interleukin-8; Luminescent Measurements; Photometry; Rats; Rats, Sprague-Dawley; Simplexvirus

Helicobacter pylori soluble proteins may serve as chemoattractants for neutrophils. Once extravasated and attracted to the gastric mucosa, neutrophils themselves may be a source of interleukin-8 (IL-8), further amplifying the inflammatory response. We evaluated IL-8 expression and the activation of human neutrophils by H. pylori products.. After neutrophils had been stimulated with H. pylori culture supernatant, IL-8 mRNA expression was assessed by quantitative reverse transcription polymerase chain reaction, using synthetic standard RNA at 0, 1, 2, 4, and 9 h. The amount of IL-8 protein was measured by enzyme-linked immunosorbent assay (ELISA). Lymphocyte function-associated antigen-1beta (LFA-1beta) (CD18) expression was determined with flow cytometry, and myeloperoxidase secretion was analyzed with ELISA. After acetohydroxamic acid (AHA) and/or N-tert-butoxycarbonyl-methionyl-leucyl-phenylalanine (BOC-MLP) was added to H. pylori culture supernatant, IL-8 ELISA was analyzed for 9 h.. IL-8 mRNA expression by stimulated neutrophils was 16 to 67 times greater than by controls, peaking at 2 h after stimulation. The amount of IL-8 protein was markedly increased at 4 h after stimulation. H. pylori culture supernatant enhanced LFA-1beta expression and myeloperoxidase secretion by neutrophils. AHA and/or BOC-MLP decreased IL-8 production at 2-4 h after stimulation.. H. pylori-induced neutrophil recruitment may be mediated via IL-8 expressed by neutrophils activated by H. pylori soluble proteins. This may explain the gastric mucosal inflammatory response to the non-invasive organism.

Topics: CD18 Antigens; Cells, Cultured; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lymphocyte Function-Associated Antigen-1; N-Formylmethionine Leucyl-Phenylalanine; Neutrophil Activation; Neutrophils; Peroxidase; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Urease

1. Helicobacter pylori infection is characterized by an infiltration of neutrophils in the gastric mucosa. Neutrophil activation is an important source of reactive oxygen radicals, which cause tissue damage. Studies have shown that in Helicobacter pylori-infected patients there is increased mucosal production of interleukin 8. However, the role of interleukin 8 in the Helicobacter pylori-related inflammatory process and its relationship with reactive oxygen radicals remains to be clarified. The aims of this study were to investigate if there is any association between antral mucosal levels of interleukin 8 and reactive oxygen radicals and their relationship to gastric antral inflammation. 2. Fifty-two patients referred for endoscopy were recruited into the study. Gastric antral biopsies were taken for histology, culture and measurement of interleukin 8 and chemiluminescence (measuring reactive oxygen radicals). Interleukin 8 was measured by ELISA and the result expressed as pg/mg biopsy. Luminol-enhanced chemiluminescence was measured as mV min-1 mg-1 biopsy. Antral inflammation was assessed by a pathologist in a blinded fashion. 3. Antral mucosal levels of interleukin 8 and reactive oxygen radicals were significantly higher in Helicobacter pylori-colonized mucosa than in Helicobacter pylori-negative mucosa. After the eradication of Helicobacter pylori in patients with duodenal ulcer the median values (ranges) of interleukin 8 and reactive oxygen radicals fell from 1.21 (0.10-2.40) to 0.65 (0.00-1.60) and from 110.0 (10.0-959.0) to 14.5 (0.0-85.0) respectively. There was a positive correlation between interleukin 8 concentration and chemiluminescence response in the antral mucosa (r = 0.72). A higher interleukin 8 concentration was associated with greater neutrophil infiltration (r = 0.72) and mononuclear cell infiltration (r = 0.55); the magnitude of the chemiluminescence response was also positively associated with neutrophil (r = 0.77) and mononuclear cell infiltration (r = 0.59). 4. Interleukin 8 concentration is associated with an infiltration of neutrophils and mononuclear cells and is correlated with the production of reactive oxygen radicals in antral gastric mucosa infected with Helicobacter pylori. These findings suggest that interleukin 8 may be important in attracting and activating phagocytes to release reactive oxygen radicals, thereby causing mucosal damage.

Topics: Duodenal Ulcer; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Interleukin-8; Pyloric Antrum; Reactive Oxygen Species

To clarify the pathological functions of the virulence factors of Helicobacter pylori, a comparative analysis was carried out on the relationship between motility, flagellar gene polymorphism, vacuolating cytotoxin (VT) production and interleukin-8 (IL-8) induction.. Twenty-five strains were examined for restriction fragment length polymorphism (RFLP) of the flagellin gene. Motility was measured using semisolid agar plates. Cytotoxicity was assayed using RK-13 cells. IL-8 secretion was assessed by the enzyme-linked immunosorbent assay (ELISA) methods.. H. pylori was classified into four groups according to their flagellar RFLP. No differences were noted in motility or VT production among the four groups, but a significant difference was noted in IL-8 induction. In addition, highly motile strains produced more IL-8.. This flagellar genetic polymorphism may be associated with IL-8 induction.

Topics: Animals; Bacterial Proteins; Bacterial Toxins; Cytotoxins; DNA Primers; DNA, Bacterial; Electrophoresis, Agar Gel; Enzyme-Linked Immunosorbent Assay; Flagellin; Gastric Mucosa; Genetic Variation; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Rabbits; Stomach Diseases; Tumor Cells, Cultured; Virulence

Lactoferrin (Lf) is an iron-binding glycoprotein present in milk, lacrimae, saliva, and gastroduodenal secretions. In vitro studies disclosed contradicting results regarding the relation of Lf with Helicobacter pylori (HP) infection. This study aimed to investigate the relationship between the gastric mucosal concentration of Lf and HP infection of the stomach. The relationship of the gastric mucosal level of Lf with the gastric mucosal concentration of interleukin-8 (IL-8) and with the intragastric ammonia levels was also assessed. In addition, the gastric mucosal Lf levels before and after irradication of HP infection were also evaluated.. This study was composed of 27 HP-positive and 12 HP-negative patients with chronic gastritis. Gastric mucosal biopsy specimens were obtained from all subjects by endoscopy, and the degree of histological inflammatory changes were assessed according to the Sydney system. The gastric mucosal levels of Lf and IL-8 were measured by immunoassays. Assessment of the effect of therapy on the gastric mucosal level of Lf was performed in 10 patients with HP-associated duodenal ulcer.. Lf, IL-8, and ammonia levels were significantly higher in patients with HP-positive gastritis compared with those with HP-negative gastritis in both the antrum and the gastric body. Histologically, the degree of inflammatory changes correlated significantly with the Lf levels in the gastric mucosa. Furthermore, the degree of HP colonization was more significant in biopsy samples from the antrum than in those from the corpus of the stomach. The gastric mucosal levels of Lf and IL-8 correlated significantly in the antrum and the gastric body. The ammonia intragastric level significantly correlated with the mucosal Lf level in the antrum and in the gastric body. Therapy significantly decreased the Lf levels in the gastric mucosa of the antrum (p < 0.005) and the gastric body (p < 0.005).. The results of the present investigation showed, for the first time in vivo, that Lf concentration is increased in the biopsy specimens of patients with HP-related gastritis, and that the levels of Lf correlate significantly with the degree of inflammation of the gastric mucosa. The gastric mucosal level of Lf may constitute an excellent marker of HP infection.

Topics: 2-Pyridinylmethylsulfinylbenzimidazoles; Ammonia; Anti-Bacterial Agents; Anti-Ulcer Agents; Biomarkers; Biopsy; Chronic Disease; Clarithromycin; Duodenal Ulcer; Female; Follow-Up Studies; Gastric Juice; Gastric Mucosa; Gastritis; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lactoferrin; Lansoprazole; Male; Metronidazole; Middle Aged; Omeprazole; Pyloric Antrum

Acute Helicobacter pylori infection produces predominantly neutrophilic infiltration of the gastric mucosa. However, the precise mechanisms and mediators of neutrophil migration are not known. Interleukin-8 (IL-8), a potent chemotactic factor for neutrophils, is present at high concentration in the gastric mucosa of subjects with chronic gastritis caused by H. pylori infection. The aims of this study were to determine whether IL-8 stimulates polymorphonuclear leukocyte (PMN) migration across a cultured monolayer of rabbit gastric epithelial cells and whether PMN migration affects epithelial cell barrier function. Confluent gastric epithelial monolayers grown on the inserts were overlaid with PMNs and various amounts of IL-8 were administered into the well under the insert. Gastric epithelial barrier function was assessed by sodium back diffusion. IL-8 stimulated PMN migration across the monolayer in a dose- and time-dependent manner. PMN transmigration significantly increased sodium back diffusion. In conclusion, IL-8 induces PMN migration across a monolayer of cultured gastric epithelial cells. This IL-8 action is associated with impairment of gastric epithelial barrier function. Since H. pylori infection causes a local mucosal increase of IL-8, our present findings may explain the mechanism of H. pylori-induced PMN infiltration of the gastric glands and mucosal injury.

Topics: Animals; Cells, Cultured; Chemotaxis, Leukocyte; Epithelial Cells; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Neutrophils; Rabbits; Recombinant Proteins

Despite the fact that Helicobacter pylori is known to be non-invasive, mucosal infiltration of inflammatory cells have been observed in the gastric mucosa. The exact pathogenesis of such an inflammatory reaction has not been well defined. We explored the repertoire of cytokine genes expressed in human gastric epithelial cells in response to coculture with H. pylori. After gastric epithelial cells, SNU-5 and KATO III, were infected with H. pylori, expression of several cytokine genes was assessed using quantitative reverse transcription polymerase chain reaction. Interleukin (IL)-8, -1 alpha and -1 beta mRNA were expressed in both gastric epithelial cells throughout the entire infection period. In SNU-5, IL-1 alpha and IL-8 mRNA were expressed at 1 h, reached a peak level at 4 h and then decreased. Interleukin-1 beta mRNA was expressed less frequently than IL-1 alpha, or IL-8 mRNA. In SNU-5 cells, granulocyte-macrophage colony-stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), and tumour necrosis factor-alpha (TNF-alpha) mRNA were expressed at 9 h, but was not expressed in KATO III. Gene expression paralleled the amount of IL-8 protein measured by enzyme-linked immunoabsorbent assay (ELISA). Interleukin-8 mRNA expression was not observed in KATO III cells infected with Campylobacter fetus ssp. fetus, Campylobacter jejuni or Escherichia coli. IL-8 mRNA expression was increased not only in gastric epithelial cells but also in non-gastric cells infected with H. pylori. These results suggest that an inflammatory reaction induced by H. pylori may be initially triggered by an array of pro-inflammatory cytokines expressed by infected gastric epithelial cells.

Topics: Campylobacter Infections; Cells, Cultured; Chemokine CCL2; Coculture Techniques; Cytokines; Epithelium; Escherichia coli Infections; Granulocyte-Macrophage Colony-Stimulating Factor; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Polymerase Chain Reaction; RNA, Messenger; Stomach; Time Factors; Tumor Necrosis Factor-alpha

Helicobacter pylori is a bacterium which causes gastric inflammatory diseases. Oral inoculation of H pylori usually results in only a temporary colonisation without a successful infection in the stomach of conventional mice in which lactobacilli are the predominant indigenous bacteria.. To determine whether lactobacilli exert an inhibitory effect on colonisation by H pylori in the stomach.. The effects of H pylori on attachment to murine and human gastric epithelial cells and the H pylori mediated release of interleukin-8 (IL-8) by these cells were examined in vitro. Lactobacillus salivarius infected gnotobiotic BALB/c mice and control germ free mice were inoculated orally with H pylori to examine whether L salivarius can inhibit colonisation by H pylori.. L salivarius inhibited both the attachment and IL-8 release in vitro. H pylori could not colonise the stomach of L salivarius infected gnotobiotic BALB/c mice, but colonised in large numbers and subsequently caused active gastritis in germ free mice. In addition, L salivarius given after H pylori implantation could eliminate colonisation by H pylori.. These findings suggest the possibility of lactobacilli being used as probiotic agents against H pylori.

Topics: Animals; Antibiosis; Bacterial Adhesion; Disease Models, Animal; Gastric Mucosa; Germ-Free Life; Helicobacter Infections; Helicobacter pylori; Interleukin-8; Lactobacillus; Male; Mice; Mice, Inbred BALB C

Active Helicobacter pylori-associated gastritis is characterized by a dense mucosal infiltration with granulocytes. Since H. pylori is noninvasive, secondary signals must induce the accumulation of granulocytes. Interleukin-8 (IL-8) has been shown to play a key role in this event. Using competitive reverse transcriptase-PCR on mRNA from gastric biopsies, we could show a clear correlation between the amount of IL-8 transcripts and the activity of H. pylori gastritis. Due to the inability of the bacterium to invade host cells, the epithelial layer is a potential candidate as an IL-8 source. To study the mechanism of IL-8 induction, established gastric carcinoma epithelial cell lines (AGS and Kato III) and well-defined H. pylori strains were used in a modified in vitro system. The experimental design enabled us to prevent direct contact of bacteria with epithelial cells by use of a filter membrane which did not block secreted bacterial products crossing the membrane. The data clearly showed that the direct contact of the bacterial cell with the epithelial cell is necessary for optimal IL-8 production because not only live bacteria, but also metabolically inactive bacteria, increased IL-8 secretion. Neither purified lipopolysaccharide nor water-soluble protein fractions of H. pylori NCTC 11637 and Tx30a nor the cytotoxin of H. pylori was able to increase IL-8 production significantly by the epithelial cells used. Furthermore, preparations of total membrane and outer membrane proteins of H. pylori were not able to stimulate IL-8 release in vitro. Accumulatively, these results imply that active metabolism is not necessary for stimulation as long as there is an intact membrane aiding the presentation of a stimulating membrane complex or aggregate on the surface of the bacteria. From these results, we conclude that whole bacteria and their direct contact with epithelial cells may be critical for IL-8 induction in vivo.

Topics: Bacterial Adhesion; Bacterial Outer Membrane Proteins; Bacterial Proteins; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; RNA, Messenger; Transcription, Genetic

Helicobacter pylori adheres to gastric epithelial cells and stimulates interleukin (IL)-8 production. This may be instrumental in neutrophil infiltration of the gastric epithelium that characterizes H. pylori gastritis. This study examined the molecular mechanisms leading to H. pylori-induced epithelial cell IL-8 production.. Electrophoretic mobility shift analyses for NF-kappa B were performed on cell and nuclear extracts from H. pylori-infected AGS and Kato III human gastric epithelial cells.. H. pylori infection activated the transcription factor NF-kappa B and induced nuclear translocation of both NF-kappa B p50/p65 heterodimers and p50 homodimers. Nuclear translocation of NF-kappa B (30 minutes) was followed by increased IL-8 messenger RNA (1 hour) and protein levels (4 hours) consistent with NF-kappa B up-regulation of IL-8 gene transcription. Pretreatment of AGS cells with PDTC, which blocks NF-kappa B activation, inhibited H. pylori-induced increases in IL-8 production by 90%. Immunohistochemical studies using a monoclonal antibody that recognizes the I-kappa B binding region of p65 showed activated NF-kappa B in gastric epithelial cells of patients with H. pylori gastritis.. H. pylori infection activates NF-kappa B in gastric epithelial cells in vitro and in vivo. NF-kappa B is a transcriptional regulator of IL-8 production, and its activation after bacterial infection may be an important defense response in gastrointestinal epithelial cells.

Topics: Biopsy; Cell Line; Cell Nucleus; Gastric Mucosa; Genes, MHC Class I; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; NF-kappa B; Recombinant Proteins; RNA, Messenger; Transcription, Genetic

Helicobacter pylori strains possessing the cagA gene are thought to induce interleukin 8 (IL-8) in gastric mucosa. However, it is still unclear whether a relation exists between the cagA gene and the expression patterns of cytokines other than IL-8.. To investigate the relation between the cagA gene and the production of various cytokine proteins using an enzyme linked immunosorbent assay (ELISA).. In 184 patients, the cagA gene was detected by polymerase chain reaction (PCR), and levels of production of IL-1 beta, IL-6, IL-7, IL-8, IL-10, and tumour necrosis factor alpha (TNF-alpha) in antral biopsy specimens were measured by ELISA.. Mucosal levels of IL-1 beta, IL-6, IL-8, and TNF-alpha were significantly higher in H pylori positive than in H pylori negative patients. Furthermore, the mucosal levels of IL-1 beta and IL-8 were significantly higher in specimens infected with cagA positive strains than in those infected with cagA negative strains. In H pylori positive patients, the mucosal level of IL-8 was closely correlated with that of IL-1 beta (p < 0.0001), and the mucosal level of IL-6 was closely correlated with that of TNF-alpha (p < 0.0001).. These findings suggest that the ability to induce cytokines differs among the strains; cagA+ strains induce various kinds of cytokines and may cause severe inflammation, whereas cagA- strains induce IL-8 and IL-1 beta only weakly and may cause only mild inflammation. However, as most patients infected with the cagA+ strains have gastritis, these strains may not be equivalent to ulcerogenic strains.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Bacterial; Bacterial Proteins; Colony Count, Microbial; Cytokines; Dyspepsia; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Gastritis; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

To clarify the relationship between interleukin-8 (IL-8) production and virulent factors, we examined the motility and cytotoxicity of H. pylori, suggested to be a major cause of chronic gastritis and peptic ulcers. Our results demonstrated that among cytotoxic strains of H. pylori, high-motility strains induced more IL-8 than low-motility strains. There was no correlation between cytotoxicity and motility of H. pylori. Four restriction fragment length polymorphism (RFLP) patterns were observed in the flaA PCR products. There was no correlation between flaA RFLP and motility. In conclusion, our findings suggest that both cytotoxicity and motility are virulent factors in the pathogenesis of gastric mucosal injury.

Topics: Animals; Cell Line; Flagellin; Gastric Mucosa; Genes, Bacterial; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Rabbits

To determine the mechanisms of gastric mucosal injury associated with Helicobacter pylori infection, we investigated the contents of cytokines and inflammatory cell infiltration in the gastric mucosa. Ninety-six patients with dyspepsia were studied (58 gastric ulcer, 38 nonulcer dyspepsia). Of the 96 patients, 63 were infected with H. pylori as determined by microscopic examination with HE staining, culture of H. pylori, or the rapid urease test. Endoscopic biopsy specimens were obtained from both the antrum and the body to examine interleukin (IL)-8, IL-6, IL-1 beta, and tumor necrosis factor-alpha contents in the gastric mucosa by enzyme-linked immunosorbent assay. Inflammatory cell infiltration was assessed according to the Sydney system. IL-8 content was enhanced in both the antral and body mucosa of the H. pylori-positive patients compared with the H. pylori-negative patients. Furthermore, IL-8 content correlated well with the infiltration of both mononuclear cells and polymorphonuclear cells. These results suggest that IL-8 plays important roles in the pathogenesis of gastric mucosal injury associated with H. pylori infection.

Topics: Case-Control Studies; Cytokines; Dyspepsia; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Stomach Ulcer

We examined secretion, mRNA expression, and histologic localization of interleukin-8 (IL-*) and growth-related gene product-alpha (GRO alpha) in the Helicobacter pylori-infected gastric antral mucosa. Antral biopsies were obtained from an area of endoscopically intact mucosa. Significantly higher levels of IL-8 and GRO alpha were secreted in organ cultures from patients with H. pylori infection, and their elevation was prominent in patients with duodenal ulcer. There was a significant association between these alpha-chemokine levels and histologic grades of activity, inflammation, and H. pylori density. In fresh antral biopsies, IL-8 and GRO alpha mRNA expression was detected more frequently in H. pylori-infected patients compared with those without infection. Immunofluorescence microscopy showed localization of IL-8 and GRO alpha proteins in gastric epithelial cells and infiltrating CD68+ macrophages. In the chemotaxis assay, a significant positive correlation was found between neutrophil migration induced by the organ culture supernatants and their contents of IL-8 and GRO alpha. After H. pylori eradication, a significant decrease was observed in IL-8 and GRO alpha levels detected in organ cultures. In conclusion, mucosal alpha-chemokine activity correlates well with histologic severity of H. pylori-associated antral gastritis and can be used to predict the effects of H. pylori eradication therapy.

Topics: Adult; Biopsy; Chemokine CXCL1; Chemokines; Chemokines, CXC; Chemotactic Factors; Chemotaxis, Leukocyte; Female; Fluorescent Antibody Technique, Indirect; Gastric Mucosa; Gastritis; Gene Expression; Growth Substances; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Male; Middle Aged; Neutrophils; Peptic Ulcer; Polymerase Chain Reaction; RNA, Messenger

Neutrophil accumulation plays an important role in Helicobacter pylori-associated gastric mucosal injury. In this study, the mucosal content of myeloperoxidase (MPO), which is a measure of neutrophil accumulation and interleukin-8 (IL-8) was assayed and changes in MPO and IL-8 content were determined before and after H. pylori eradication therapy. Thirty-seven H. pylori-positive patients (11DU/26GU) underwent H. pylori eradication therapy with lansoprazole (30 mg/day, 6 weeks) and amoxicillin (2 g/day, 2 weeks), followed by famotidine (20 mg/day, 8 weeks). H. pylori-infection status was evaluated by routine endoscopic examinations (culture, CLO, histology). Immediately and 8 weeks after cessation of the anti-H. pylori therapy, these tests were repeated. Intragastric urease activity was estimated by delta 13CO2, which was obtained by the [13C]urea breath test (UBT). Mucosal samples were taken and tissue MPO and IL-8 contents were assayed by EIA and ELISA, respectively. Histologic examination was also performed. Among the 37 patients, 21 cases of H. pylori infection were eradicated (56.8%). Intragastric urease activity was dramatically reduced immediately after the anti-H. pylori therapy, whereas, it was re-elevated 8 weeks later in the relapsed cases. Antral MPO content was decreased in the eradicated and relapsed cases. MPO in the corpus was also decreased in the eradicated cases. Nevertheless, it was enhanced (3.5-fold) in the relapsed cases at 8 weeks after therapy. Changes in mucosal IL-8 content were similar to those of MPO. In eradicated cases, neutrophil infiltration is improved in both the antrum and corpus. However failure of eradication therapy results in the enhancement of neutrophil accumulation in the corpus. Further study is necessary to clarify the mechanism of neutrophil accumulation after therapy for H. pylori.

Topics: 2-Pyridinylmethylsulfinylbenzimidazoles; Amoxicillin; Anti-Bacterial Agents; Anti-Ulcer Agents; Biopsy; Drug Therapy, Combination; Duodenal Ulcer; Famotidine; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Lansoprazole; Male; Middle Aged; Neutrophils; Omeprazole; Peroxidase; Proton Pump Inhibitors; Stomach Ulcer

To examine the background histology, interleukin-8 (IL-8) secretion, and expression of IL-8 mRNA and protein, using the gastric antral mucosa infected with Helicobacter pylori.. The antral biopsies were obtained from an area of endoscopically intact mucosa in 147 patients whose endoscopic diagnoses were normal (n = 41), duodenal ulcer (n = 58), gastric ulcer (n = 21), or gastritis (n = 27). Levels of IL-8 secreted in the organ cultures of mucosal biopsies were measured by an ELISA assay, and the expression of IL-8 mRNA and protein was analyzed in fresh biopsy tissues with RT-PCR and immunofluorescent microscopy, respectively.. Significantly greater levels of IL-8 were secreted in patients with H. pylori infection, and its elevation was more prominent in duodenal ulcer patients than in those with gastric ulcer or endoscopically defined gastritis. There was an association among H. pylori density, IL-8 activity, and histological severity of activity and inflammation of gastritis in the Sydney system. Consistent with enhanced IL-8 activity in the organ cultures, IL-8 mRNA was detected in 16 of 23 fresh biopsy tissues studied in H. pylori-positive patients. In contrast, IL-8 transcript was detected in only one of 12 H. pylori-negative cases. Immunofluorescent microscopy showed localization of IL-8 protein in the gastric epithelial cells and lamina propria cells, primarily CD68+ macrophages in specimens with H. pylori infection.. This study indicates that a strong correlation exists between mucosal IL-8 activity and histological severity in H. pylori-associated antral gastritis. Further studies will be necessary to determine the mechanisms involved in elevated mucosal IL-8 activity in H. pylori infection.

Topics: Adult; Biopsy; Duodenal Ulcer; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Organ Culture Techniques; Pyloric Antrum; RNA, Messenger; Stomach Ulcer

Within the gastroduodenal mucosa Helicobacter pylori infection stimulates local production of a range of proinflammatory and immunoregulatory cytokines, neutrophil infiltration, specific T- and B-cell responses and the development of gastric lymphoid follicles. Following bacterial eradication this mucosal inflammatory response resolves. Infiltrating neutrophils are likely to be one of the major mediators of mucosal damage. Neutrophil activation, including reactive oxygen metabolite production and the release of myeloperoxidase, will be induced directly by bacterial factors and indirectly through products of complement activation, bioactive lipids and host-derived cytokines. Interleukin-8, and related peptides of the chemokine family secreted by gastric epithelial cells, are likely to be important host mediators inducing neutrophil migration to sites of infection. Epithelial IL-8 is upregulated by TNF-alpha and IL-1 and directly by H. pylori strains expressing the CagA phenotype. The extent of mucosal injury may reflect bacterial density, the variability of different strains of H. pylori to induce chemokine expression in epithelial cells and the oxidative burst in neutrophils. Recent evidence from in vivo and in vitro studies shows that CagA+ VacA+ strains of H. pylori are associated with enhanced inflammatory responses and mucosal damage. Defining the specific bacterial mediators of mucosal inflammation will be important in elucidating the role of H. pylori in the pathogenesis of gastroduodenal disease.

Topics: Antigens, Bacterial; Bacterial Proteins; Bacterial Toxins; Chemotaxis, Leukocyte; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-12; Interleukin-8; Neutrophil Activation; Neutrophils; Species Specificity

To assess its clinicopathological and diagnostic significance, interleukin-8 (IL-8) was measured by radioimmunoassay in fasting urine specimens and in gastric mucosal incubates taken from 54 patients with dyspepsia. The presence of Helicobacter pylori, the activity of gastritis, and urine creatinine levels were also assessed.. The median urinary IL-8/creatinine ratio was 0.1 x 10(-6) in patients with current peptic ulcers (n = 13) and 0.2 x 10(-6) in patients with a history of ulcers (n = 8), compared with 0.4 x 10(-6) (p < 0.0001) in patients without ulcers who were infected with H. pylori (n = 20) or not infected (n = 13). The activity of gastritis had a positive correlation with gastric IL-8 (r = 0.5870, p < 0.01) and a negative correlation with urinary IL-8/creatinine ratio (r = -0.6447, p < 0.005). The improvement in the activity of gastritis in 20 patients given anti-H. pylori triple therapy was associated with a significant fall in gastric mucosal IL-8 and a rise in urinary IL-8/creatinine ratio.. An inverse relationship seems to exist between urinary IL-8 and the activity of gastritis and gastric IL-8. This may represent another concept in the pathogenesis of peptic ulcers and can assist in the noninvasive diagnosis of peptic ulcer disease.

Topics: Adult; Creatinine; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Peptic Ulcer; Prospective Studies; Radioimmunoassay

Following exposure to Helicobacter pylori cells, epithelial cell lines secreted interleukin-6 (IL-6) and IL-8 but not tumor necrosis factor alpha. Purified IL-6 alone did not stimulate IL-8 production from the cell lines tested, indicating that IL-6 was not an intermediary in IL-8 induction. Enhanced IL-8 secretion occurred in a time- and dose-dependent manner. None of 12 antibiotics tested exhibited a significant effect on IL-8-inducing activity, suggesting that preformed antigens were responsible for stimulating IL-8 secretion in vitro. Live bacterial cells caused the highest level of stimulation. Proteinase-digested and heated (56 or 100 degrees C) cells had significantly reduced stimulatory activities. Purified H. pylori lipopolysaccharide, but not exopolysaccharide, stimulated low-level secretion of IL-8, but only at high concentrations, while a water-extracted H. pylori antigen preparation was strongly stimulatory for HEp-2 cells. No reduction in IL-8-stimulatory activity was observed for H. pylori mutants negative for urease activity, production of a major lipoprotein, and motility. The noncytotoxic strain CCUG 915 stimulated lower IL-8 levels than other isolates. However, the otherwise isogenic cytotoxin-negative mutant 17874 delta vacA (S. H. Phadnis, D. Ilver, L. Janzon, S. Normark, and T. U. Westblom, Infect. Immun. 62:1557-1565, 1994) had the same IL-8-stimulatory ability as the parent strain, suggesting that surface proteins other than the vacuolating cytotoxin are involved in IL-8 stimulation.

Topics: Bacterial Proteins; Bacterial Toxins; Cytokines; Cytotoxins; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Epithelium; Helicobacter Infections; Helicobacter pylori; Humans; Immunoblotting; Interleukin-8; Polymerase Chain Reaction; Polysaccharides, Bacterial; RNA, Messenger; Tumor Cells, Cultured

To investigate the role of interleukin-8 (IL-8) and tumour necrosis factor (TNF) in patients infected with Helicobacter pylori.. The study population comprised 52 patients with dyspepsia attending for upper gastrointestinal endoscopy. Of these patients, 35 were infected with H pylori. IL-8 and TNF concentrations in plasma, gastric juice, and gastric biopsy homogenate supernatant fluid were measured by radioimmunoassay and L929 cell bioassay, respectively.. The concentrations of IL-8 and TNF in gastric juice and gastric biopsy homogenates were substantially greater in patients infected with H pylori. In H pylori positive patients IL-8 concentrations in gastric juice and gastric biopsy homogenates were higher in those with moderate gastritis than in those with mild gastritis. There was a positive correlation between IL-8 and TNF concentrations in gastric juice and gastric biopsy homogenate supernatant fluid from H pylori positive patients. There were no significant differences between H pylori positive and negative patients with respect to IL-8 and TNF plasma concentrations.. This study suggests that increased gastric production of IL-8 and TNF may be implicated in the pathogenesis of H pylori associated gastroduodenal disease.

Topics: Adult; Aged; Biological Assay; Cytokines; Dyspepsia; Female; Gastric Juice; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

Helicobacter pylori is associated with neutrophil infiltrates, although the mechanism of their recruitment is only partially defined. The aim of the study was to determine if Kato III, a human gastric epithelial cell line, expressed cytokines and the intercellular adhesion molecule 1 (ICAM-1), which could contribute to the initiation of inflammation during infection with H. pylori.. Kato III cells were stimulated with H. pylori and were examined for evidence of infection, cytokine production, and the expression of ICAM-1.. The expression of interleukin 8 messenger RNA and immunoreactive protein by Kato III cells was significantly increased over constitutive levels within 3 hours of infection with H. pylori. Infected Kato III supernatants activated neutrophils as evidenced by increased CD11b/CD18 and decreased L-selectin that could be blocked by anti-interleukin 8. In contrast, Campylobacter jejuni, lipopolysaccharide, killed H. pylori, and supernatants from cultures of H. pylori did not increase interleukin 8. Interleukins 2 and 6; interferons alfa, beta, and gamma; and tumor necrosis factor were not produced by resting or H. pylori-stimulated Kato III cells. In addition to producing interleukin 8, Kato III constitutively expressed surface ICAM-1, which acts as an intercellular adhesion molecule for neutrophils.. Our results indicate that H. pylori stimulates the gastric epithelium to initiate inflammation and neutrophil recruitment and activation.

Topics: Base Sequence; Campylobacter Infections; Campylobacter jejuni; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Molecular Probes; Molecular Sequence Data; RNA, Messenger; Salmonella typhimurium; Tumor Cells, Cultured

Infection of the gastric antrum by Helicobacter pylori is characterised by a cellular inflammatory infiltrate. Whether cytokines are involved in the pathogenesis of this gastritis has been investigated by studying the effect of eradicating H pylori on the expression of genes encoding the cytokines interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF-alpha) in the antral mucosa. Gastric antral biopsy specimens were taken from nine patients with duodenal ulcers and cytokine transcripts were identified and quantified by northern blotting. After H pylori had been eradicated the chronic inflammatory infiltrate decreased in all the patients and the polymorphonuclear infiltrate virtually disappeared. Expression of genes also decreased. After eradication, the median TNF-alpha mRNA/rRNA fell to 48% (p = 0.02) and the median IL-8 mRNA/rRNA fell to 5% (p = 0.004) of initial values. These results support the role of increased synthesis of these cytokines in the pathogenesis of the gastritis.

Topics: Adult; Aged; Base Sequence; Blotting, Northern; Duodenal Ulcer; Female; Follow-Up Studies; Gastritis; Gene Expression; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Male; Middle Aged; Molecular Sequence Data; Pyloric Antrum; RNA, Messenger; Tumor Necrosis Factor-alpha

We investigated whether tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-8 (IL-8) are involved in the inflammatory reaction of Helicobacter pylori infection. In 23 patients with H. pylori infection and 16 patients with negative cultures for H. pylori and normal antral mucosa, the mucosal production of TNF-alpha, IL-1 beta, and IL-8 was measured in antral biopsy specimens after 23 h of in vitro culture. The levels of TNF-alpha and IL-1 beta appeared to be significantly higher in H. pylori-positive patients (p = 0.0002 for both TNF-alpha and IL-1 beta). IL-8 production was also higher in H. pylori-infected subjects, but this difference did not reach statistical significance (p = 0.057). No significant differences were found between the level of the cytokines in H. pylori-infected patients with or without duodenal ulcer disease. A strong correlation was found between the production of IL-1 beta and IL-8. The biologic effects of these cytokines may explain the conspicuous recruitment, influx, and activation of neutrophils in the gastric mucosa during H. pylori infection.

Topics: Adult; Aged; Enzyme-Linked Immunosorbent Assay; Female; Gastric Mucosa; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

To investigate the expression of interleukin-8 (IL-8) in Helicobacter pylori infected normal and neoplastic gastroduodenal mucosa, and in established gastric cancer cell lines.. Immunofluorescence techniques were used to localise IL-8 in cryosections of gastric (n = 25) and duodenal (n = 17) endoscopic biopsy specimens an in resected gastric tumour tissue samples from 16 patients. Two gastric cancer cell lines (Kato 3 and MKN 45) were examined for IL-8 protein expression by immunofluorescence and for the presence of IL-8 mRNA by reverse transcription followed by the polymerase chain reaction (RT-PCR).. IL-8 was localised to the epithelium in histologically normal gastric mucosa, with particularly strong expression in the surface cells. IL-8 expression was also a feature of surface epithelium in the duodenal bulb, but was much reduced in the second part of the duodenum. In chronic H pylori-associated gastritis gastritis gastric epithelial IL-8 expression was increased and expression of IL-8 within the lamina propria was evident. By contrast, large areas of IL-8 negative epithelium were observed in the body mucosa of a subject with Ménétrier's disease. In gastric carcinoma the tumour cells were positive for IL-8. IL-8 was also detected by immunofluorescence in unstimulated Kato 3 and MKN 45 cells, and constitutive IL-8 gene expression in these cell lines was confirmed by detection of IL-8 mRNA by RT-PCR.. Immunoreactive IL-8, a potent neutrophil chemotactic and activating factor, is present in the epithelium of both normal and inflamed gastric mucosa with increased expression in the latter. There is site dependent variation in epithelial IL-8 expression within the gastroduodenal mucosa. The expression of the pro-inflammatory cytokine IL-8 in gastric carcinoma cells may influence peritumoural cellular infiltrates.

Topics: Base Sequence; Chronic Disease; Duodenitis; Duodenum; Fluorescent Antibody Technique; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-8; Intestinal Mucosa; Molecular Sequence Data; RNA, Neoplasm; Stomach Diseases; Stomach Neoplasms; Tumor Cells, Cultured

To determine the concentrations of interleukin-1 beta, interleukin-6, and interleukin-8 in tissue homogenates of mucosal biopsy specimens from Helicobacter pylori-positive and -negative patients.. In 43 consecutive patients who underwent upper gastrointestinal endoscopy, seven antral biopsies were taken; three specimens were used for cytokine determination and the remaining four biopsies were processed for H. pylori detection. Peripheral venous blood was collected and IgG to H. pylori was assayed by an ELISA technique.. Twenty-nine of 43 patients (67%) were histologically positive for H. pylori; all had chronic gastritis. The mucosal levels of interleukin-6 and interleukin-8 were significantly higher in H. pylori-positive patients than in the negative patients (p < 0.001). A significantly higher percentage of interleukin-8 was found in patients colonized by H. pylori with active superficial chronic gastritis (85.7%), compared to quiescent superficial gastritis (12.5%) (p < 0.01), and the median and range were, respectively, 400 (0-1000) and 0 (0-200) pg/mg protein (p < 0.001). In patients with active superficial gastritis, a significant correlation between interleukin-6 and -8 was found (p 0.01). No difference was found regarding the mucosal levels of interleukin-1 beta according to the presence of H. pylori.. These results suggest a possible pathogenetic role for interleukin-6 and interleukin-8 in H. pylori-associated gastritis.

Topics: Adult; Aged; Chronic Disease; Dyspepsia; Female; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged

Gastric infection with Helicobacter pylori is frequently characterized by neutrophil infiltration. The production of the neutrophil-activating peptide (NAP-1/IL-8) and mucosal IgA autoantibodies to IL-8 by human antral biopsies have been examined during short-term in vitro culture. Detectable IL-8 was secreted by 84% of H. pylori-negative patients with normal antral mucosa (range < 0.07-61.5 ng/mg biopsy protein, n = 19). Concentrations in 4 patients with reactive gastritis and 10 with inactive gastritis were not significantly different from subjects with normal mucosa. In H. pylori-positive patients with active gastritis and neutrophil infiltration into the epithelium (n = 17) IL-8 secretion was significantly increased relative to subjects with normal mucosa (P < 0.0001), inactive gastritis (P < 0.001) and reactive gastritis (P < 0.01). IL-8 concentrations in active gastritis were significantly correlated with the extent of epithelial surface degeneration (r = 0.64). IgA autoantibodies were present in 19 patients (13 active, 4 inactive gastritis) and concentrations were significantly correlated with IL-8 production (P < 0.001). Gastric synthesis of IL-8 is likely to be an important factor in regulating mucosal neutrophil infiltration and activation in patients with H. pylori infection. The local production of IgA antibodies to IL-8 may represent a down-regulatory response of the host to limit mucosal damage associated with a chronic bacterial infection.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Autoantibodies; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Immunoglobulin A, Secretory; Interleukin-8; Middle Aged