Seroreactivity to specific antigens of Helicobacter pylori infection is associated with an increased risk of the dyspeptic gastrointestinal diseases

SummaryObjectivesThe correlation between seroreactivity to Helicobacter pylori-specific antigens and clinical outcomes in gastrointestinal disease remains unresolved. We investigated the anti-H. pylori antibody profile in northeast Thai dyspeptic patients with gastrointestinal disease in order to identify any H. pylori antigens that may be associated with an increased risk of gastrointestinal disease.Patients and methodsEighty-nine H. pylori-infected dyspeptic patients (44 non-ulcer, 23 peptic ulcer, 22 gastric cancer) were included in the study. Patients were considered to have H. pylori infection when at least one invasive method (i.e., culture, rapid urease test, and histology on biopsy specimens) and serological tests including a commercial ELISA (Pyloriset EIA-GIII) and a commercial immunoblot (Helicoblot 2.1; Genelabs Diagnostics), were positive. In addition, the sera of 20 H. pylori-infected blood donors and 10 H. pylori-non-infected blood donors were also randomly collected and analyzed for H. pylori infection by ELISA and Helicoblot 2.1.ResultsImmunoreactive protein bands at 116-kDa, 89-kDa, 37-kDa, 35-kDa, 30-kDa, 19.5-kDa, and the current infection marker for H. pylori-infected patients had average frequencies of 97.8%, 77.5%, 36.0%, 25.8%, 79.8%, 58.4%, and 69.7%, respectively. The immunoreactive patterns obtained from the H. pylori-infected patients and H. pylori-infected blood donors were similar. The antibodies to VacA and CagA antigens were not significantly different among the H. pylori-infected gastroduodenal patient groups. The simultaneous presence of antibody to 19.5-kDa antigen and absence of antibody to 35-kDa antigen was associated with an increased risk of gastric cancer (p<0.05). The immunoreactive band to 35-kDa antigen was found at significantly higher levels in peptic ulcer patients, and the 37-kDa antigen was found at significantly higher levels in non-ulcer patients (both p<0.05). Significantly low levels of antibodies to 23-kDa and 85-kDa antigens were found associated with peptic ulcer (p<0.05).ConclusionsWe confirm that the universal presence of CagA and VacA in H. pylori-infected patients in Thailand is independent of the gastroduodenal disease. The presence or absence of antibodies to H. pylori-specific antigens may be useful as indirect markers in the screening of H. pylori-infected patients, and may have specific protection roles in H. pylori-related gastroduodenal diseases

Helicobacter pylori in Thai patients with cholangiocarcinoma and its association with biliary inflammation and proliferation

AbstractObjectivesTo investigate whether Helicobacter spp. infection and the cagA of H. pylori are associated with hepatobiliary pathology, specifically biliary inflammation, cell proliferation and cholangiocarcinoma (CCA).MethodsHelicobacter species including H. pylori, H. bilis and H. hepaticus were detected in the specimens using the polymerase chain reaction (PCR). Biliary inflammation of the liver and gallbladders was semi-quantitatively graded on hematoxylin and eosin (H&E)-stained slides. Biliary proliferation was evaluated by immunohistochemistry using the Ki-67-labelling index.ResultsHelicobacter pylori was found in 66.7%, 41.5% and 25.0% of the patients in the CCA, cholelithiasis and control groups (P < 0.05), respectively. By comparison, H. bilis was found in 14.9% and 9.4% of the patients with CCA and cholelithiasis, respectively (P > 0.05), and was absent in the control group. The cagA gene of H. pylori was detected in 36.2% and 9.1% of the patients with CCA and cholelithiasis, respectively (P < 0.05). Among patients with CCA, cell inflammation and proliferation in the liver and gallbladder were significantly higher among those DNA H. pylori positive than negative.ConclusionsThe present findings suggest that H. pylori, especially the cagA-positive strains, may be involved in the pathogenesis of hepatobiliary diseases, especially CCA through enhanced biliary cell inflammation and proliferation

Helicobacter pylori cag pathogenicity island (cagPAI) involved in bacterial internalization and IL-8 induced responses via NOD1- and MyD88-dependent mechanisms in human biliary epithelial cells

Helicobacter pylori infection has been proposed to be associated with various diseases of the hepatobiliary tract, including cancer of the bile duct epithelial cells (cholangiocarcinoma, CCA). The ability of H. pylori bacteria to cause pathogenic effects in these cells has, however, yet to be investigated. Given that the cag pathogenicity island (cagPAI) is required for H. pylori pathogenesis in gastric epithelial cells, we investigated wild-type and cag mutant strains for their ability to adhere, be internalized and induce pro-inflammatory responses in two bile duct epithelial cell lines derived from cases of CCA. The findings from these experiments were compared to results obtained with the well-characterized AGS gastric cancer cell line. We showed that the cagPAI encodes factors involved in H. pylori internalization in CCA cells, but not for adhesion to these cells. Consistent with previous studies in hepatocytes, actin polymerization and α5β1 integrin may be involved in H. pylori internalization in CCA cells. As for AGS cells, we observed significantly reduced levels of NF-κB activation and IL-8 production in CCA cells stimulated with either cagA, cagL or cagPAI bacteria, when compared with wild-type bacteria. Importantly, these IL-8 responses could be inhibited via either pre-treatment of cells with antibodies to α5β1 integrins, or via siRNA-mediated knockdown of the innate immune signaling molecules, nucleotide oligomerization domain 1 (NOD1) and myeloid differentiation response gene 88 (MyD88). Taken together, the data demonstrate that the cagPAI is critical for H. pylori pathogenesis in bile duct cells, thus providing a potential causal link for H. pylori in biliary tract disease

3697/N

Abstract. Helicobacter pylori, an important etiological agent in the development of gastritis, peptic ulcer and gastric carcinoma, can be detected by the enzyme-linked immunosorbent assay (ELISA). Our objectives were: 1) to evaluate the efficacy of a commercial ELISA kit (Pyloriset EIA-G III) in sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for diagnosis of H. pylori infection in Thai dyspeptic patients in Khon Kaen Thailand; and 2) to examine the seroprevalence of H. pylori among blood donors at Srinagarind Hospital&apos;s Blood Bank, Khon Kaen University, by the commercial ELISA. Gastric biopsies obtained from 137 dyspeptic patients were diagnosed by culture, rapid urease test (RUT) and histology. Serum samples from the same dyspeptic patients and 100 healthy blood donors were assayed using the commercial ELISA. H. pylori infection in dyspeptic patients was considered positive when the culture or both RUT and histology were positive. Using a cut-off value at a titer of 20 U/ml (as recommended by the manufacturer), we found the commercial ELISA kit had a sensitivity of 93.3%, specificity of 75.3%, PPV of 74.7%, NPV of 93.5% and accuracy of 83.2%. The overall H. pylori seroprevalence in the healthy blood donors was 57%. Of the 100 healthy blood donors, 39 (60.9%) of the males and 18 (50.0%) of the females were seropositive

Actin polymerization and <i>H. pylori</i> internalization.

<p><b>A</b>. <b>Effect of cytochalasin D (actin polymerization inhibitor) on <i>H. pylori</i> internalization in biliary (KKU-100 and KKU-M156) and gastric cells (AGS)</b>. After treatment with cytochalasin D, cells were incubated with <i>H. pylori</i> wild type, <i>cagA</i> or <i>cag</i>PAI mutant strains for 6 h. The <i>H. pylori</i> internalization was assessed by bacterial culture. The percentage of <i>H. pylori</i> internalization in cytochalasin D-treated cells was compared to the number of <i>H. pylori</i> internalization in untreated control cells<b>. B</b>. <b>Effect of α5β1 integrin antibodies on <i>H. pylori</i> internalization in biliary (KKU-100 and KKU-M156) and gastric (AGS) cells</b>. After pre-treatment with α5β1 integrin antibodies, cells were incubated with <i>H. pylori</i> wild type, cagA<b>^-</b> or <i>cag</i>PAI<b>^-</b> strains for 6 h. <i>H. pylori</i> internalization was accessed by bacterial culture. The percentage of <i>H. pylori</i> internalization in α5β1 integrin-antibody-treated cells was compared to the number of <i>H. pylori</i> internalization in untreated control cells. Data are the mean ± SEM of triplicate experiments. * <i>p</i> < 0.05 represented a significant difference compared between cytochalasin D or α5β1 integrin antibody-treated cells and untreated cells.</p

<i>H. pylori</i> up-regulates <i>NOD1</i>, <i>TLR4</i> and <i>TLR5</i> gene expression in biliary cells.

<p><b>A</b>. <b><i>NOD1</i>, B. <i>TLR4</i> and C. <i>TLR5</i> gene expression in KKU-100 and AGS cells after stimulation with <i>H. pylori</i> wild type, cagA^-, <i>cag</i>L^- or <i>cag</i>PAI^- strains for 6, 12 and 24 h</b>. Gene expression was accessed by qRT-PCR. Data are presented as the mean ± SEM of triplicate experiments. * <i>p</i> < 0.05, ** <i>p</i> < 0.01 represent a significant difference between control, non-stimulated cells and <i>H. pylori</i>-stimulated cells. ^#<i>p</i> < 0.05, ^##<i>p</i> < 0.01 represent a significant difference between <i>H. pylori</i> wild type- and <i>cag</i>PAI^- -stimulated cells.</p

The carcinogenic liver fluke Opisthorchis viverrini is a reservoir for species of Helicobacter

There has been a strong, positive correlation between opisthorchiasis-associated cholangiocarcinoma and infection with Helicobacter. Here a rodent model of human infection with Opisthorchis viverrini was utilized to further investigate relationships of apparent co-infections with O. viverrini and H. pylori. A total of 150 hamsters were assigned to five groups: i) Control hamsters not infected with O. viverrini; ii) O. viverrini-infected hamsters; iii) non-O. viverrini infected hamsters treated with antibiotics (ABx); iv) O. viverrini-infected hamsters treated with ABx; and v) O. viverrini-infected hamsters treated both with ABx and praziquantel (PZQ). Stomach, gallbladder, liver, colonic tissue, colorectal feces and O. viverrini worms were collected and the presence of species of Helicobacter determined by PCR-based approaches. In addition, O. viverrini worms were cultured in vitro with and without ABx for four weeks, after which the presence of Helicobacter spp. was determined. In situ localization of H. pylori and Helicobacter-like species was performed using a combination of histochemistry and immunohistochemistry. The prevalence of H. pylori infection in O. viverrini-infected hamsters was significantly higher than that of O. viverrini-uninfected hamsters (). Interestingly, O. viverrini-infected hamsters treated with ABx and PZQ (to remove the flukes) had a significantly lower frequency of H. pylori than either O. viverr-iniinfected hamsters treated only with ABx or O. viverrini-infected hamsters, respectively (). Quantitative RT-PCR strongly confirmed the correlation between intensity H. pylori infection and the presence of liver fluke infection. In vitro, H. pylori could be detected in the O. viverrini worms cultured with ABx over four weeks. In situ localization revealed H. pylori and other Helicobacter-like bacteria in worm gut. The findings indicate that the liver fluke O. viverrini in the biliary tree of the hamsters harbors H. pylori and Helicobacter-like bacteria. Accordingly, the association between O. viverrini and H. pylori may be an obligatory mutualism

<i>H. pylori</i> induces IL-8 production in biliary cells through NOD1 and MyD88 signaling pathways.

<p><b>A</b>. <b>IL-8 production, B. <i>NOD1</i> and C. <i>MYD</i>88 gene expression in KKU-100 cell line and AGS cells treated with NOD1 or MyD88 siRNA then stimulated for 24 h with <i>H. pylori</i> wild type or <i>cag</i>PAI^- strains</b>. Cells transfected with siRNA to the HBD3 gene (<i>DEFB10</i>3) were used as a negative control. Cells without bacteria are indicated as non-stimulated. IL-8 production was determined by ELISA. Data are presented as the mean ± SEM of triplicate experiments. ** <i>p</i> < 0.01 represents significant differences between non-stimulated and <i>H. pylori</i>-stimulated cells. ^##<i>p</i> < 0.01 represents a significant difference between the control and NOD1 or MyD88 siRNA-treated cells. Data are presented as the mean ± SEM of triplicate experiments. ^#<i>p</i> < 0.05 represents a significant difference between the non-stimulated and <i>H. pylori</i>-treated cells (white bar). * <i>p</i> < 0.05, ** <i>p</i> < 0.001 represents a significant difference between the control and NOD1 or MyD88 siRNA-treated cells.</p