Western Blotting of Total Lysate of Helicobacter pylori in Cases of Atrophic Body Gastritis

Abstract Background: Atrophic body gastritis is considered the first important step in the histogenesis of gastric carcinoma, a multistep process starting from chronic gastritis and progressing through chronic atrophic gastritis, intestinal metaplasia, and dysplasia. Helicobacter pylori is involved in the induction of atrophic body gastritis, but documentation of H. pylori infection is difficult because of the progressive disappearance of the bacterium. Our study aimed to detect past H. pylori infection in patients with atrophic body gastritis. Methods: We used Western blot analyses of whole bacterial protein lysate of 2 different strains to probe sera from 143 patients. All sera were analyzed by ELISA (Bio-Rad), and results of gastric histology were available for all patients. Results: Among 111 patient sera previously classified as negative for H. pylori infection by ELISA, 106 (95.5%) were positive when assayed by immunoblotting. Conclusions: Commercial diagnostic reagent sets may fail to detect H. pylori infection. Western blotting of whole bacterial protein extracts could provide the basis of a noninvasive serology tool able to assess previous infection with H. pylori in patients with atrophic body gastritis.

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Analysis of Helicobacter pylori vacA andcagA genotypes and serum antibody profile in benign and malignant gastroduodenal diseases

Gut ◽

10.1136/gut.43.2.182 ◽

1998 ◽

Vol 43 (2) ◽

pp. 182-186 ◽

Cited By ~ 59

Author(s):

D Basso ◽

F Navaglia ◽

L Brigato ◽

M G Piva ◽

A Toma ◽

...

Keyword(s):

Helicobacter Pylori ◽

Gastric Adenocarcinoma ◽

Western Blotting ◽

Serum Antibody ◽

Serological Response ◽

Benign Diseases ◽

Antral Gastritis ◽

Antibody Profile ◽

H Pylori ◽

Different Strains

Background—Helicobacter pylori species comprise different strains, cytotoxic and non-cytotoxic, which can be identified on the basis of their genomic pattern.Aims—(1) To evaluate the polymorphism of the vacA gene and to ascertain whether thecagA gene is present in patients with gastric adenocarcinoma. (2) To study the anti-H pylori antibody profile using western blotting.Patients—Twenty one patients with gastric adenocarcinoma and 71 with H pyloriassociated benign disease (nine gastric ulcer, 29 duodenal ulcer, 25 antral gastritis, and eight duodenitis).Methods—The polymerase chain reaction was used to verify the presence or absence ofcagA and to study the polymorphism of vacA in gastric mucosal samples obtained during endoscopy for patients with benign diseases and at surgery for patients with gastric adenocarcinoma. Fasting sera were used to assess anti-H pylori antibodies against different H pyloriantigens by western blotting.Results—cagAgene and the allele s1 of vacAwere significantly less frequent in patients with antral gastritis (60% and 60%) compared with patients with gastric adenocarcinoma (94% and 100%) and with other non-malignant gastroduodenal diseases (93% and 87%) (χ2=16.01, p<0.001; and χ2=13.97, p<0.01). In patients with gastric adenocarcinoma, antibodies against a 74 kDa H pylori antigen were less frequently found than in patients with benign diseases.Conclusions—H pylori infection caused bycagApositive/vacA s1 strains is a frequent finding in patients with gastric adenocarcinoma. Prospective studies are needed to confirm whether the low incidence of positive serological response to the 74 kDa H pyloriantigen in patients with gastric adenocarcinoma is important.

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Features of the endoscopic picture in paediatric gastroduodenal diseases caused by Helicobacter pylori

Russian Journal of Infection and Immunity ◽

10.15789/2220-7619-fot-1249 ◽

2020 ◽

Vol 10 (3) ◽

pp. 543-550

Author(s):

Sh. T. Turdieva ◽

E. A. Shamansurova

Keyword(s):

Helicobacter Pylori ◽

Inflammatory Diseases ◽

Gastric Wall ◽

Duodenogastric Reflux ◽

Chronic Atrophic Gastritis ◽

Endoscopic Examination ◽

Ulcer Disease ◽

Functional Studies ◽

H Pylori ◽

Endoscopic Picture

The aim of the study was to examine the features of the endoscopic picture in the upper digestive tract mucosa in paediatric chronic gastroduodenal pathology (CGDP) associated with Helicobacter pylori. Materials and methods. There were examined 286 patients, aged 6–15 years. Diagnostic criteria for chronic gastroduodenal pathology were anamnestic as well as instrumental and functional studies data: gastric fractional intubation, esophagogastroduodenoscopy (EPGDS) with endoscopic pH-metry without biopsy, and ultrasound examination of abdominal organs. H. pylori testing was carried out by two unrelated methods such as respiratory test and a immunochromatographic fecal test. Results. Detection of H. pylori in children with CGDP peaked in patients with peptic gastric and duodenal ulcer (up to 87.5%, p < 0.05). The main endoscopic signs were edema, hyperaemia and contact bleeding, as well as local haemorrhage were the major endoscopic signs of inflammation in the stomach and duodenum mucosa. Atrophic mucosal lesions were characterized by thinning, pale colour together with transilluminated submucosal vessels. Non-atrophic antral gastritis was featured with delayed gastric emptying, antral stasis and pyloric spasm. In contrast, hypotension of the gastric wall, duodenogastric reflux and decreased motility were more typical to chronic atrophic gastritis. Major endoscopic feature in patients with H. pylori infection was presented by dominant atrophic changes combined with gastroduodenal reflux (77.6%, p < 0.05) compared to patients without H. pylori infection. Conclusion. Detection of HP infection was peaked in children with CGDP coupled to peptic ulcer disease compared to patients with inflammatory diseases (p < 0.05). Endoscopic examination in HP-positive patients showed that atrophic changes were found by 4-fold more frequently together with gastroduodenal reflux compared to patients without HP infection (p < 0.05).

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Therapeutic effects of Zuojin Pill on Helicobacter pylori-induced chronic atrophic gastritis through JMJD2B/COX-2/VEGF signaling axis

10.21203/rs.3.rs-20652/v2 ◽

2020 ◽

Author(s):

Shihua Wu ◽

Chunmei Bao ◽

Ruilin Wang ◽

Xiaomei Zhang ◽

Sijia Gao ◽

...

Keyword(s):

Helicobacter Pylori ◽

Cell Viability ◽

Atrophic Gastritis ◽

Chronic Atrophic Gastritis ◽

Gastric Mucosal Damage ◽

Therapeutic Effects ◽

Cox 2 ◽

H Pylori

Abstract Background: Zuojin Pill (ZJP), a famous Chinese medicinal formula, widely accepted for treatment of chronic atrophic gastritis (CAG) in China. This study aimed to explore the therapeutic effects and mechanisms of ZJP in Helicobacter pylori (H. pylori) - induced chronic atrophic gastritis (CAG) in vivo and in vitro. Methods: CAG rat model was induced by H. pylori. ZJP (0.63, 1.26, and 2.52 g/kg, respectively) was administered orally for four weeks. Therapeutic effects of ZJP were identified by H&E staining and serum indices. In addition, cell viability, morphology and proliferation were detected by cell counting kit-8 (CCK8) and high-content screening assay (HCS), respectively. Moreover, relative mRNA expression and protein expression related to JMJD2B/COX-2/VEGF axis was detected to investigate the potential mechanisms of ZJP in CAG. Results: Results showed the symptoms (weight loss and gastric mucosa damage) of CAG were alleviated, and the contents of TNF-α in serum was markedly decreased after treating with ZJP. Moreover, cell viability, proliferation and morphology changes of GES-1 cells were ameliorated by ZJP intervention. In addition, proinflammatory genes and JMJD2B/COX-2/VEGF axis related genes were suppressed by ZJP administration in vitro and in vivo. Meanwhile, immunohistochemistry (IHC) and western blot confirmed down-regulation of these genes by ZJP intervention. Conclusion: ZJP treatment can alleviate gastric mucosal damage induced by H. pylori via JMJD2B/COX-2/VEGF axis.

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Are Histopathological Changes of H. pylori Infection in Young Dyspeptic Patients Necessitate Endoscopy?

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2019.725 ◽

2019 ◽

Vol 7 (19) ◽

pp. 3211-3215

Author(s):

Wafaa Redha Mohammed Al-Sabbagh ◽

Alaa Qasim Yahya ◽

Rasha Abdelraouf Alsafi

Keyword(s):

Helicobacter Pylori ◽

Gastric Lymphoma ◽

Young Patients ◽

Chronic Atrophic Gastritis ◽

Gastric Biopsy ◽

P Value ◽

Histopathological Changes ◽

Gastric Biopsies ◽

Antigen Tests ◽

H Pylori

BACKGROUND: Helicobacter pylori is an important gastrointestinal infective bacteria with many serious complications including gastric erosions and ulceration, duodenal ulcer, gastric carcinoma and MALT gastric lymphoma. The gastric biopsy is commonly performed in H. pylori-positive dyspeptic individuals, and many previous researchers studied the histopathological features of infected gastric biopsies however little previous studies focused on the histopathological findings in young population in comparison to the older one. AIM: To make a focus on the histopathological effects of H. pylori infection in young patients compared with the older one and predicts the need for endoscopy in this population, also to estimates the prevalence of infection in Iraqi patients. MATERIAL AND METHODS: the sample for this study is 180 patients in total, they attended Marjan medical city in Iraq for dyspepsia of more than 3 months and prepared for OGD. Patients asked for their permission to do immunological tests for H. pylori. Both serology for H. pylori antibodies and stool for antigen tests are used, and the case is included in the study only if both tests were positive, after OGD, the gastric biopsies are processed and examined histopathologically. RESULTS: Normal gastric biopsy is the most common histopathological finding in young (< 25 years) patients (75%) while chronic atrophic gastritis is the most common one in patients > 25 years age (57%). The prevalence of Helicobacter pylori infection in dyspeptic patients was 73.3%, the correlation between infection and sex was insignificant (p-value 0.06), and no significant correlation between infection and age (p-value 0.07) was concluded. CONCLUSION: H. pylori-related histopathological changes of gastric mucosa in young (< 25 years) are commonly mild and does not necessitate endoscopy at this age unless there are alarming signs.

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Serum Antibody Responses to Helicobacter pylori and the cagA Marker in Patients with Mucosa-Associated Lymphoid Tissue Lymphoma

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.6.4.633-638.1999 ◽

1999 ◽

Vol 6 (4) ◽

pp. 633-638 ◽

Cited By ~ 8

Author(s):

Anne Taupin ◽

Alessandra Occhialini ◽

Agnès Ruskone-Fourmestraux ◽

Jean-Charles Delchier ◽

Jean-Claude Rambaud ◽

...

Keyword(s):

Helicobacter Pylori ◽

Antibody Response ◽

Malt Lymphoma ◽

Lymphoid Tissue ◽

Serum Antibody ◽

Dot Blot ◽

Homologous Strain ◽

H Pylori ◽

Different Strains ◽

Dot Blot Analysis

ABSTRACT The lymphoma of the mucosa-associated lymphoid tissue (MALT) of the stomach has been linked to Helicobacter pylori infection, but the mechanisms involved in B-cell proliferation remain elusive. In a search for putative H. pylori-specific monoclonal immunoglobulin production, an H. pylori strain was isolated from 10 patients with MALT lymphoma and used to detect the specific serum antibody response to the homologous strain by immunoblotting. Moreover, the antigenicity of the different strains was compared by using each of the 10 sera. We found that the different strains induced highly variable patterns of systemic immunoglobulin G antibody response, although several bacterial antigens, such as the 60-kDa urease B, were often recognized by the different sera. ThecagA marker was detected in the strains by PCR with specific primers and by dot blot analysis, and the CagA protein was found in the sera of 4 of the 10 patients by immunoblotting. In conclusion, MALT lymphoma patients, like other patients with H. pylori gastritis, exhibit a polymorphic systemic antibody response, despite an apparently similar antigenic profile. The CagA marker of pathogenicity is not associated with this disease.

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Detection and variability analyses of CRISPR-like loci in the H. pylori genome

10.7287/peerj.preprints.27196v1 ◽

2018 ◽

Author(s):

Jerson Alexander Garcia-Zea ◽

Roberto de la Herrán ◽

Francisca Robles Rodríguez ◽

Rafael Navajas-Pérez ◽

Carmelo Ruiz Rejón

Keyword(s):

Helicobacter Pylori ◽

Geographical Area ◽

Pathogenic Bacterium ◽

Gene Location ◽

Phylogenetic Marker ◽

Genomic Plasticity ◽

Human Pathogenic Bacterium ◽

H Pylori ◽

Different Strains ◽

Genomic Locations

Helicobacter pylori is a human pathogenic bacterium with a high genomic plasticity. Although the functional CRISPR-Cas system has not been found in its genome, CRISPR like loci have been recently identified. In this work, 53 genomes from different geographical areas are analyzed for the search and analysis of variability of this type of structure. We confirm the presence of a locus that was previously described in the VlpC gene in al lgenomes, and we characterize new CRISPR-like loci in other genomic locations. By studying the variability and gene location of these loci, the evolution and the possible roles of these sequences are discussed. Additionally, the usefulness of this type of sequences as a phylogenetic marker has been demonstrated, associating the different strains by geographical area.

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Association of Helicobacter pylori infection and stomach cancer: our experience

International Surgery Journal ◽

10.18203/2349-2902.isj20183193 ◽

2018 ◽

Vol 5 (8) ◽

pp. 2794

Author(s):

N. G. Javan ◽

Wormi Sharon

Keyword(s):

Gastric Cancer ◽

Helicobacter Pylori ◽

Gastric Carcinoma ◽

Stomach Cancer ◽

Histopathological Examination ◽

Chronic Atrophic Gastritis ◽

Helicobacter Pylori Infection ◽

Future Research ◽

Gastric Cancer Prevention ◽

H Pylori

Background: Infection with Helicobacter pylori (H. pylori) has been linked with chronic atrophic gastritis, an inflammatory precursor of gastric adenocarcinoma. There are data on the epidemiology, pathophysiology, and histology of this disease that show that Helicobacter pylori gastritis has an important role in gastric carcinogenesis. However, it has to be considered that only very few of those infected with Helicobacter pylori will develop gastric cancer. Hence, it will be a major target of future research to identify individuals who carry a greater risk for developing gastric cancer, and therefore may benefit from eradication of Helicobacter pylori in terms of gastric cancer prevention. Various studies revealed that approximately more than 50% of the world’s human population is infected by Helicobacter pylori. In underdeveloped countries, this association is shown to be much higher according to different studies.Methods: This study was conducted over a period of 36 months from 1st January 2014 till December 31st, 2016. All patients who underwent Gastrectomy during this period were taken. All specimens were investigated to see presence of helicobacter pylori by histological examination. A total of 50 Gastrectomy was performed by one surgical team over 36-month period.Results: Out of 50 patients, Helicobacter pylori positivity was seen in 33 (66%) cases by histopathological examination (HPE). Gastric cancer is more prevalent among males 31 (62%) as compared to 19 (38%) in females. It is more common among the older age group.Conclusions: Helicobacter pylori infection is higher in prevalence in cases of stomach cancer. Present study also showed that there is significant association of Helicobacter pylori infection with gastric carcinoma. Helicobacter pylori infection could be one of the etiological factors for gastric carcinoma.

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Where to Biopsy to Detect Helicobacter pylori and How Many Biopsies Are Needed to Detect Antibiotic Resistance in a Human Stomach

Journal of Clinical Medicine ◽

10.3390/jcm9092812 ◽

2020 ◽

Vol 9 (9) ◽

pp. 2812 ◽

Cited By ~ 1

Author(s):

Maxime Pichon ◽

Cong Tri Tran ◽

Gaëtan Motillon ◽

Charlotte Debiais ◽

Sylvain Gautier ◽

...

Keyword(s):

Helicobacter Pylori ◽

Dna Typing ◽

Resistant Isolate ◽

Subtotal Resection ◽

Routine Practice ◽

Gene Detection ◽

Antibiotic Resistant ◽

The Third ◽

H Pylori ◽

Different Strains

This study aims to determine the gastric distribution, density, and diversity of Helicobacter pylori infection. Subtotal resection of the stomachs of three H. pylori-infected and asymptomatic obese patients were collected after a sleeve gastrectomy. Distribution and density of H. pylori were determined using culture and RT-PCR on multiple gastric sites (88, 176, and 101 biopsies per patient). Diversity of H. pylori strains was studied using antibiotic susceptibility testing, random amplified polymorphism DNA (RAPD) typing and cagA gene detection on single-colony isolates (44, 96, and 49 isolates per patient). H. pylori was detected in nearly all analyzed sites (354/365 biopsies, 97%). Antral density was higher in one patient only. The three stomachs were almost exclusively infected by an antibiotic-susceptible strain. One clarithromycin-resistant isolate in one biopsy was detected in two stomachs (1/44 and 1/49 isolates), while in the third one, eight (8/96 isolates) metronidazole-resistant isolates were detected. DNA typing showed infection with cagA-negative strains for one patient, cagA-positive strains for a second patient and the third patient was infected with two different strains of distinct cagA genotypes. Infection with H. pylori is shown to spread to the whole surface of the stomach, but a possibility of minor sub-population of antibiotic-resistant clones, undetectable in routine practice.

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Heterogeneity among Helicobacter pylori Strains in Expression of the Outer Membrane Protein BabA

Infection and Immunity ◽

10.1128/iai.72.6.3429-3435.2004 ◽

2004 ◽

Vol 72 (6) ◽

pp. 3429-3435 ◽

Cited By ~ 52

Author(s):

Ewa E. Hennig ◽

Ray Mernaugh ◽

Jennifer Edl ◽

Ping Cao ◽

Timothy L. Cover

Keyword(s):

Helicobacter Pylori ◽

Membrane Protein ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Recombinant Antibody ◽

Wild Type ◽

Single Chain ◽

H Pylori ◽

Different Strains

ABSTRACT The BabA adhesin of Helicobacter pylori is an outer membrane protein that binds to the fucosylated Lewis b histo-blood group antigen on the surface of gastric epithelial cells. We screened a phage-displayed ScFv (single-chain fragment variable) recombinant antibody library for antibodies reactive with a recombinant BabA fragment and identified two such antibodies. Each antibody recognized an ∼75-kDa protein present in wild-type H. pylori strain J99 but absent from an isogenic babA mutant strain. An immunoreactive BabA protein was detected by at least one of the antibodies in 18 (46%) of 39 different wild-type H. pylori strains and was detected more commonly in cagA-positive strains than in cagA-negative strains. Numerous amino acid polymorphisms were detected among BabA proteins expressed by different strains, with the greatest diversity occurring in the middle region of the proteins. Among the 18 strains that expressed a detectable BabA protein, there was considerable variation in the level of binding to Lewis b in vitro. Heterogeneity among H. pylori strains in expression of the BabA protein may be a factor that contributes to differing clinical outcomes among H. pylori-infected humans.

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Phase Variation in Helicobacter pylori Lipopolysaccharide

Infection and Immunity ◽

10.1128/iai.66.1.70-76.1998 ◽

1998 ◽

Vol 66 (1) ◽

pp. 70-76 ◽

Cited By ~ 101

Author(s):

B. J. Appelmelk ◽

B. Shiberu ◽

C. Trinks ◽

N. Tapsi ◽

P. Y. Zheng ◽

...

Keyword(s):

Helicobacter Pylori ◽

Lipid A ◽

Main Chain ◽

Phase Variation ◽

Polymeric Form ◽

Altered Expression ◽

Core Lipid ◽

Parental Phenotype ◽

H Pylori ◽

Different Strains

Helicobacter pylori NCTC 11637 lipopolysaccharide (LPS) expresses the human blood group antigen Lewis x (Lex) in a polymeric form. Lex is β-d-galactose-(1-4)-[α-l-fucose-(1-3)]-β-d-acetylglucosamine. Schematically the LPS structure is (Lex) n -core-lipid A. In this report, we show that Lex expression is not a stable trait but that LPS displays a high frequency (0.2 to 0.5%) of phase variation, resulting in the presence of several LPS variants in one bacterial cell population. One type of phase variation implied the loss of α1,3-linked fucose, resulting in variants that expressed nonsubstituted polylactosamines (also called the i antigen), i.e., Lex minus fucose; LPS: (lactosamine) n -core-lipid A. The switch of Lex to i antigen was reversible. A second group of variants arose by loss of polymeric main chain which resulted in expression of monomeric Ley; LPS: (Ley)-core-lipid A. A third group of variants arose by acquisition of α1,2-linked fucose which hence expressed Lex plus Ley; LPS: (Ley)(Lex) n -core-lipid A. The second and third group of variants switched back to the parental phenotype [(Lex) n -core-lipid A] in lower frequencies. Part of the variation can be ascribed to altered expression levels of glycosyltransferase levels as assessed by assaying the activities of galactosyl-, fucosyl-, andN-acetylglucosaminyltransferases. Clearly phase variation increases the heterogeneity of H. pylori, and this process may be involved in generating the very closely related yet genetically slightly different strains that have been isolated from one patient.

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