oipA Gene Research Articles

Influence of oipA Phase Variation on Virulence Phenotypes Related to Type IV Secretion System in Helicobacter pylori.

oipA, an outer membrane protein of Helicobacter pylori, is linked to IL-8 induction and gastric inflammation, but its role is debated due to inconsistent findings. This study aims to explore the role of oipA phase variation in modulating the virulence traits of H. pylori, a bacterium strongly associated with the development of gastric cancer. American clinical isolate AH868 strain for naturally occurring phase variations of the oipA gene, and G27 strain for invitro-induced phase variations were used to elucidate oipA's impact on key virulence phenotypes, including cell elongation, CagA phosphorylation, and IL-8 induction. Using AH868 strain, natural oipA phase variation does not affect cell elongation and IL-8 induction. Interestingly, however, invitro-induced oipA phase variations in G27 strain uncovered that 9.4% of oipA "Off" transformants exhibit reduced cell elongation while all maintaining consistent IL-8 induction levels. Additionally, complementation of oipA "Off to On" status restores the cell elongation phenotype in 12.5% of transformants, highlighting the importance of oipA in maintaining normal cell morphology. Crucially, these variations in cell elongation are not linked to changes in bacterial adherence capabilities. Furthermore, the study shows a correlation among oipA phase variation, cell elongation, and CagA phosphorylation, suggesting that oipA influences the functionality of the Type IV secretion system. Whole-genome sequencing of selected transformants reveals genetic variations in bab paralogue, cagY gene, and other genomic regions, underscoring the complex genetic interactions that shape H. pylori's virulence. Our research provides new insights into the subtle yet significant role of oipA phase variation in H. pylori pathogenicity, emphasizing the need for further studies to explore the intricate molecular mechanisms involved. This understanding could pave the way for targeted therapeutic strategies to mitigate the impact of H. pylori on human health.

HELICOBACTER PYLORI OIPA VIRULENCE GENE AS A MOLECULAR MARKER OF SEVERE GASTROPATHIES.

Helicobacter pylori is an etiologic agent of gastroduodenal diseases. The microorganism, considered a type I carcinogen, affects about 50% of the global population. H. pylori virulence factors are determinant for the clinical outcome of the infection. The outer inflammatory protein A (oipA) gene encodes an outer membrane adhesin and is related to severe gastropathies, such as gastric cancer. The aim of this study was to evaluate the association of the oipA gene with the severity of gastroduodenal diseases in dyspeptic patients in region Central Brazil. The polymerase chain reaction (PCR) was used to determine the presence of H. pylori. Samples positives were used for molecular screening of the oipA gene. Gastropathies were categorized as non-severe and severe diseases. Approximately 68% of patients had H. pylori and 36% were infected with H. pylori oipA+ strains. Infection was significantly associated in patients aged over 44 years (P=0.004). However, there was no association between oipA and patients' age (P=0.89). Approximately 46% of patients infected with oipA+ strains had some severe illness. Gastric adenocarcinoma was the most frequent severe gastropathy. The H. pylori oipA genotype was inversely associated with the severity of gastroduodenal diseases (OR=0.247, 95%CI: 0.0804-0.7149 and P=0.007). The characterization of possible molecular markers will contribute to personalized medicine, impacting the prognosis of patients. • Evidence points to an association between the H. pylori oipA gene and gastropathies. • There is a high prevalence of H. pylori infection with a relevant percentage of oipA+ strains. • More severe gastropathies were observed in those infected with H. pylori oipA+ strains.

Effect of the switch status of Helicobacter pylori outer inflammatory protein A on gastric diseases

Helicobacter pylori OipA (Outer Inflammatory Protein A) is an outer membrane protein that takes role in the adherence and colonization to the stomach. oipA gene expression is regulated by the slipped-strand mispairing mechanism through a hypermutable CT dinucleotide repeat motif in the 5΄ region. Alterations in the CT number repeats cause frame-shift mutations to result in phase variation of oipA expression. While a functional “On” status has been recognized as a risk factor for peptic ulcer diseases and gastric cancer in many studies, some controversial findings still exist. To this end, this study compiled the sequence data of oipA from 10 different studies between 2000–2019 and 50 oipA DNA sequences from our own research that examined the relationship between the phase On/Off status of oipA and gastric diseases based on CT repeat number. Overall, we have reached 536 oipA DNA sequences from patients. This large collection of oipA sequences first clarified the absolute conservation of the peptide-pentamer of FWLHA for phase ‘’On’’ status, suggesting this pentamer as a superior marker for the determination of oipA status than counting the number of CT repeats. Combining the sequence and patient data, we have re-analyzed the association between the ‘‘On’’ status of oipA and gastric diseases. Our results showed a strong association between oipA ‘‘On’’ status and gastric cancer supporting previous findings. We also investigated the AlphaFold2 computed structure of OipA that adopts a beta-barrel fold closely resembling to the autotransporter family of H. pylori. Altogether, this study confirms a strong association between oipA ‘‘On’’ statuses and severe gastrointestinal diseases like cancer and provides useful insights into the FWLHA pentamer as an indicator of “On” status of oipA putative autotransporter function rather than CT repeats number.

Genetic diversity of the oipA gene among Helicobacter pylori isolates and clinical outcome in Vietnam.

Outer inflammatory protein A (OipA), which is encoded by the oipA gene, can induce interleukin-8 secretion in gastric epithelial cells. The functional status of the oipA gene is regulated by the slipped-strand mispairing mechanism based on the CT dinucleotide repeat number in the 5' region. This study aimed to investigate the oipA functional status ("on/off") of Helicobacter pylori (H. pylori) and its association with gastroduodenal diseases in southwestern Vietnam. The cross-sectional study was conducted on 173H. pylori isolates from 173 patients with gastroduodenal diseases. Sanger sequencing was used to determine the functional status of oipA. Multivariable logistic regression analysis was performed to identify the association between oipA status and gastroduodenal diseases. The oipA "on" status accounted for 96% of H. pylori isolates. Twenty-five CT repeat patterns of the oipA 5' signal region were observed, five of which were novel CT repeat patterns. The oipA "on" status was found in 100%, 97.8%, and 86.8% of H. pylori isolates from patients with peptic ulcer, precancerous lesions, and chronic gastritis, respectively (p<0.01). The oipA "on" status was related to gastric precancerous lesions versus chronic gastritis (adjusted OR=7.39, 95% CI: 1.35-40.59, p=0.021) and peptic ulcers versus chronic gastritis (adjusted OR=12.79, 95% CI: 1.19-1760.32, p=0.033). Our data show a high prevalence of the oipA "on" status, which was associated with precancerous gastric lesions and peptic ulcers. Moreover, genetic diversity in the number and pattern of CT dinucleotide repeat of oipA among Vietnamese H. pylori strains was identified.

Analysis of Functional Status of Genetically Diverse OipA Gene in Indian Patients with Distinct Gastrointestinal Disease.

Helicobacter pylori (H. pylori,) a genetically diversified bacteria which colonizes human gastric epithelium, is now established causative agent for gastric cancer worldwide. Outer membrane protein (OMP)-coding genes of H. pylori are responsible for attachment and colonization of bacteria. These genes which code proteins on outer membrane of H. pylori is a group of 33 genes which with other virulent genes are causative of giving rise to disease-causing factors in the host. OipA (Outer inflammatory protein A), a participant of Hop family of OMP, is effective in acting as a biomarker for studying progression of diseases like gastric cancer. The functionality of oipA gene is regulated by phase variation within CT repeat pattern. It is the expression, i.e., "on"/"off" of oipA gene which is related with the development of distinct gastric diseases. 40 amplified DNA sequences were studied to investigate functional status of oipA. Our results reveal 57.2% isolates with functional oipA along with significant association with cagA (P = 0.0011) and vacAs1m1/s1m2 (P = 0.0034, P = 0.0093) genotypes, respectively. In conclusion, our results indicate diversity in CT repeat pattern among Indian H. pylori strains. The prevalence of functional oipA gene was found to be ranging between 50% and 64.2% though it did not show significant correlation between functional oipA and disease outcome.

Genetic Diversity and Functional Status of the oipA Gene of Helicobacter pylori. Its Association with Genes of the Pathogenicity Island in Patients with Gastroduodenal Diseases in the Republic of Belarus

Genetic Diversity and Functional Status of the oipA Gene of Helicobacter pylori. Its Association with Genes of the Pathogenicity Island in Patients with Gastroduodenal Diseases in the Republic of Belarus

Prevalence of some Helicobacter pylori virulence genes such vacA, oipA and dupA in Iraqi patients

Development of H. pylori-associated diseases is determined by a number of virulence factors. So, numerous molecular studies performed about the frequency of its virulence genes in infected patients. To evaluate the prevalence of some H. pylori virulence genes such vacA, oipA and dupA in some Iraqi infected patients. A total of 105 patients suffering from gastritis, clinical manifestations of dyspepsia or burning, vomiting, bloating and others symptoms, with age ranged 17-85 years who attended the Gastroenterology and Hepaetology Teaching Centre, Baghdad – Iraq. They were diagnosed through physicians according to endoscopic findings. Further biopsy samples from different places of stomach were collected by gastroenterologists from each patient, used for histopathological examination and genetic detection of virulence genes (vacA, dupA and oipA) by PCR technique. The result showed that histological study pointed, 78(74.2%) positive for H.pylori, then PCR technique applied on these positive samples to detect the presence of virulence genes which found, VacA gene was the most prevalent 73(93.5%) of positive samples, followed by oipA gene 42(53.8%) and dupA 25(32.05%). The study revealed a significant association between virulence genes and development of certain form of gastric infections. So, detection of the H. pylori virulence genes in gastric biopsy samples can be provides a clear prognosis of clinical outcomes.

Genetic diversity and functional status of oipA GENE Helicobacter pylori, its association with island of pathogenicity genes in patients with gastroduodenal diseases in the Republic of Belarus

<h3>ЦЕЛЬ ИССЛЕДОВАНИЯ</h3> Изучить вариабельность структурной организации гена <i>oipA</i> и его функциональный статус в образцах <i>H. pylori</i> с использованием метода секвенирования, оценить его связь с наличием генов острова патогенности <i>cagPAI</i> и характером клинического течения хеликобактериоза. <h3>МАТЕРИАЛ И МЕТОДЫ</h3> Объектом исследования были 84 образца ДНК <i>H. pylori</i>, выделенных из биоптатов антрального отдела слизистой оболочки желудка пациентов с различными гастродуоденальными заболеваниями. Детекция генов острова патогенности <i>HP</i> (<i>cagPAI</i>) была выполнена методом ПЦР. Анализ функционального статуса гена <i>oipA</i> проводили методом секвенирования. <h3>РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ</h3> Установлено, что 79,8% образцов <i>H. pylori</i> имеют статус гена <i>oipA</i> «включен», а остальные 20,2% образцов имели нефункциональный статус гена. Показано, что статус oipA «включен» достоверно ассоциирован с повышенным риском развития язвы ДПК по сравнению с поверхностным гастритом (<i>p</i>&lt;0,003, ОШ=14,5). Связи между статусом <i>oipA</i> и риском развития атрофического гастрита не выявлено. Исследование показало высокую частоту выявления генов острова патогенности среди образцов ДНК <i>H. pylori</i> со статусом гена <i>oipA</i> «включен». Наблюдается достоверная корреляция между включенным геном <i>oipA</i> и наличием всех изученных генов <i>cagPAI</i>, наибольший коэффициент корреляции отмечен для генов <i>cagT</i> и <i>cagM</i>. <h3>ЗАКЛЮЧЕНИЕ</h3> Связь между функциональным статусом гена <i>oipA</i> и факторами патогенности, кодируемыми островом патогенности <i>cagPAI</i>, играет важную роль в патогенезе <i>H. pylori</i> и требует дальнейшего изучения.

A comparative whole genome analysis of Helicobacter pylori from a human dense South Asian setting.

Helicobacter pylori, a Gram‐negative bacterium, is associated with a wide range of gastric diseases such as gastritis, duodenal ulcer, and gastric cancer. The prevalence of H pylori and risk of disease vary in different parts of the world based on the prevailing bacterial lineage. Here, we present a contextual and comparative genomics analysis of 20 clinical isolates of H pylori from patients in Bangladesh. Despite a uniform host ethnicity (Bengali), isolates were classified as being part of the HpAsia2 (50%) or HpEurope (50%) population. Out of twenty isolates, eighteen isolates were cagA positive, with two HpEurope isolates being cagA negative, three EPIYA motif patterns (AB, AB‐C, and ABC‐C) were observed among the cagA‐positive isolates. Three vacA genotypes were observed with the s1m1i1dic1 genotype observed in 75% of isolates; the s1m2i1d1c2 and s2m2i2d2c2 genotypes were found to be 15% and 10% of isolates, respectively. The non‐virulent genotypes s2m2i2d2c2 was only observed in HpEurope population isolates. Genotypic analysis of oipA gene, present in all isolates, revealed five different patterns of the CT repeat; all HpAsia2 isolates were in “ON” while 20% of HpEurope isolates were genotypically “OFF.” The three blood group antigen binding adhesins encoded genes (bab genes) examined and we observed that the most common genotype was (babA/babB/‐) found in eight isolates, notably six were HpAsia2 isolates. The babA gene was found in all HpAsia2 isolates but present in only half of the HpEurope isolates. In silico antibiotic susceptibility analysis revealed that 40% of the strains were multi‐drug resistant. Mutations associated with resistance to metronidazole, fluoroquinolone, and clarithromycin were detected 90%, 45%, and 5%, respectively, in H pylori strain. In conclusion, it is evident that two populations of H pylori with similar antibiotic profiles are predominant in Bangladesh, and it appears that genotypically the HpAisa2 isolates are potentially more virulent than the HpEurope isolates.

Outer inflammatory protein of Helicobacter pylori impacts IL-8 expression, adherence, cell apoptosis and cell cycle of gastric cells independent of its copy number.

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 24h. 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.

Prevalence of Helicobacter pylori babA, oipA, sabA, and homB genes in isolates from Chinese patients with different gastroduodenal diseases.

Disease outcome is associated with virulence factors of Helicobacter pylori (H. pylori), which are partially attributed to the outer membrane protein (OMP). This study aimed to investigate the correlation between the four OMP genes (babA, oipA, sabA, and homB) and gastroduodenal diseases. One hundred and seventy-seven H. pylori strains were isolated from Chinese patients with different gastroduodenal diseases (49 chronic gastritis, 19 gastric ulcer, 33 gastric cancer, and 76 duodenal ulcer), 94 of which contained pathological information (41 superficial gastritis, 24 intestinal hyperplasia, and 29 gastric adenocarcinoma). The full-length amplification of babA, oipA, sabA, and homB genes was acquired and sequenced. Then, the genetic polymorphism was analyzed to compare with the reference strains from the GenBank database. Functional status and cluster analysis were also performed to evaluate the impact of genetic polymorphism on disease outcome. The prevalence of babA, oipA, sabA, and homB genes were 91.5%, 100%, 94.0%, and 95.5%, respectively. The four OMP genes were characterized by genetic polymorphism and in the status of positive selection (Ka/Ks> 1). The proportion of strains with functional status on for oipA and sabA gene was 100% and 76.2%, respectively. The sequences of four OMP genes were mainly clustered together with the East Asian references. The four OMP genes were not different in patients with gastroduodenal diseases and pathologic changes (P > 0.05). H. pylori babA, oipA, sabA, and homB genes were common in the Chinese populations, but did not seem to be involved in the development of gastroduodenal diseases.

Helicobacter pylori and Epstein–Barr Virus Infection in Gastric Diseases: Correlation with IL-10 and IL1RN Polymorphism

Introduction Helicobacter pylori and Epstein–Barr virus (EBV) infection have recently been shown to be associated with gastric diseases. Polymorphisms in genes encoding cytokines such as interleukin 10 (IL-10) and interleukin 1 Receptor (IL-1RN) influence cytokine secretion levels and appear to contribute to the risk of developing gastroduodenal diseases. To our knowledge, this is the first preliminary study to address the association of coinfection with H. pylori and EBV and their correlation with genetic predisposition in the development of gastric diseases. Methods Gastric biopsy samples of 96 patients with different gastric diseases were used. Results Our results showed that the rate of coinfection was higher in patients with gastric cancer than in patients with normal gastric mucosa, active chronic gastritis, and MALT lymphoma. As regards the characterization of H. pilory strains, the polymorphism s1m1i1 of vacA gene was more frequent in patients with MALT Lymphoma in comparison to others, while the polymorphism s2m2i2 was most frequent in patients with normal gastric mucosa. In addition, patients who tested positive for the cagA gene were more frequently those affected with gastric cancer than those with inactive chronic gastritis. Similarly, the patients with oipA gene ON were more frequently those with gastric cancer than those with inactive chronic gastritis. Conclusion According to our analysis, there was no correlation between coinfection and polymorphisms in genes encoding IL-10 and IL-1RN. We conclude that various factors can be involved in the development of gastric diseases.

Co-expression of Interleukin-17A molecular adjuvant and prophylactic Helicobacter pylori genetic vaccine could cause sterile immunity in Treg suppressed mice

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.

Status (on/off) of oipA gene: their associations with gastritis and gastric cancer and geographic origins.

Virulence factors of H. pylori, such as outer inflammatory protein A (oipA), are closely involved in the development of gastric diseases such as chronic gastritis and gastric cancer. The functional status of oipA is regulated by a repair mechanism based on CT dinucleotide repeats that influence the reading frame, thus granting the gene a functional or nonfunctional status; in other words, the functional status of the oipA gene seems to be associated with the development of gastric diseases. This study sought to detect the presence of the oipA gene and to determine its functional status in patients with gastric diseases. We analyzed 516 biopsy samples (101 with normal gastric tissue, 365 with chronic gastritis, and 50 with gastric cancer). The presence of oipA was determined by PCR, and the gene status was determined using sequencing reactions. The oipA gene was found to be associated with the development of chronic gastritis, and the "on" status of the gene was the most frequent in patients with gastric cancer who were from Western countries. The CT repeats revealed geographic characteristics, but it is the functional status of the oipA gene that seems to be involved in the development of gastric diseases and in the development of gastric cancer in particular.

Genetic diversity and functional analysis of oipA gene in association with other virulence factors among Helicobacter pylori isolates from Iranian patients with different gastric diseases

Helicobacter pylori (H. pylori) is one of the most genetically diverse bacterial pathogens that persistently colonizes the human gastric epithelium. This remarkable genomic plasticity may act as a driving force for successful adaptation and persistence of the bacteria in the harsh gastric environment. Outer inflammatory protein A (OipA) encoded by oipA gene (HP0638/hopH) is a member of the outer membrane proteins (OMPs) of H. pylori involved in induction of IL-8 secretion and is associated with development of peptic ulcer and gastric cancer. Expression of OipA is regulated by phase variation within a CT dinucleotide repeat motif of the oipA gene. In this study we carried out direct DNA sequence analysis of 53 amplified fragments to investigate the oipA “On/Off” status among Iranian H. pylori isolates from patients with various gastric diseases. The prevalence of cagL, cagA, EPIYA motifs, vacA alleles, babA2 and sabA genotypes as well as cagPAI integrity of the isolates were determined by PCR. Our results demonstrated a high prevalence of strains with functional oipA status (79%) and significant associations were found between functional oipA and cagA (P = 0.027) and vacA s1m1 (P = 0.022) genotypes. The vacA s1m2 genotype was also found to be statistically associated with PUD (P = 0.0001). Interestingly, we showed that H. pylori strains with intact cagPAI co-expressed oipA gene in a significant synergistic relationship (P < 0.01). However, no significant association was observed between the functional oipA status and clinical outcomes (P > 0.05). In conclusion, our findings denotes great diversity in the number and pattern of CT dinucleotide repeats of oipA among Iranian H. pylori strains. The synergistic link between functional oipA and other important virulence factors is proposed to be critical in the pathogenesis of H. pylori, which needs further studies with a larger number of samples.

Molecular Identification and Detection of Virulent Factors in Helicobacter pylori from Gastric Biopsy Samples of Patients Attended at Assam Medical College and Hospital, Dibrugarh, Assam, India

The present study aimed to detect the major virulence determinants of Helicobacter pylori, the gastric bacteria by polymerase chain reaction from genomic DNA of 314 gastric biopsies from dyspeptic patients in 2015-2016 through upper gastrointestinal endoscopy. In 153 cases out of 314, the high prevalence of oipA gene followed by cagA-vacA s1 m1 combined genotypes was found in mostly gastritis/duodenitis patients followed by the peptic ulcer and normal patients. Therefore, the clinical significance of the virulence markers of H. pylori associated with the severe forms of gastroduodenal diseases is still a matter of controversy since the endoscopically normal patients were found to harbour the virulent genes.

Binding of the Helicobacter pylori OipA causes apoptosis of host cells via modulation of Bax/Bcl-2 levels

The H. pylori outer inflammatory protein A (OipA) is an outer membrane protein that contributes to gastric inflammation. OipA is believed to affect intra-cellular signalling and modulate the host signalling pathways. The aim of the current study was to clarify the role of OipA in H. pylori pathogenesis and its effect on host cell signalling pathways. To this end, the oipA gene was isolated and inserted into cloning and expression vectors. The recombinant plasmid was transferred into an expression host to produce OipA, which was subsequently purified by affinity chromatography and used for antibody production. A confluent monolayer of gastric cell lines was treated with various concentrations of OipA and investigated for attachment, toxicity, and apoptosis and alterations in signalling pathways. OipA bound to gastric cell lines confirming its role in the attachment of H. pylori to host cells. The ratio of Bax/Bcl-2 and caspase3, 8, FasL in the host cells were assessed and the results showed that the Bax/Bcl-2 ratio as well as the level of cleaved-caspase 3 was elevated in OipA-treated cells. These findings suggest that OipA can bind and induce toxic events as well as triggering apoptotic cascade in host gastric cells through intrinsic pathway.

Design, cloning and expression assay of oipA gene in a bicistronic vector harboring mice IL-18 gene: potential implications for Helicobacter pylori vaccine investigations

Introduction: Helicobacter pylori (H. pylori) infection has remained as a global health problem. Animal studies demonstrated the role of H. pylori oipA gene in the development of gastric cancer. The aim of this study was the cloning and expression of Helicobacter pylori oipA gene in a bicistronic vector harboring mice IL-18 gene. Materials and methods: The target gene encoding oipA was amplified from a codonoptimized clone by PCR, and then double-digested by restriction enzymes. The pIRESIgk/ mIL18/Fc plasmid was simultaneously digested by BstXI/NotI enzymes to elicit the eGFP segment. PCR product of oipA was inserted into pIRES-Igk/mIL18/Fc plasmid using T4 ligase. Transformation into DH5α strain was done. Cloning was confirmed by PCR, enzymatic digestion and sequencing. Expression of the oipA and IL-18 mRNA was assessed by means of TaqMan Real-time PCR. Results: Electrophoresis of PCR product, enzymatic digestion and sequencing showed that the H. pylori oipA gene was successfully cloned into pIRES-Igk/mIL18/Fc to generate mIL- 18-pIRES2-oipA plasmid. The results of Real-time PCR confirmed the successful expression of both oipA and IL-18 in mouse macrophage cell line. Conclusion: Considering the role of oipA in pathogenesis of H. pylori and potent activity of IL-18 as a molecular adjuvant, the results of the present study showed that the expression of codon-optimized oipA gene in bicistronic vector including mouse IL-18 is successful. So, it could be considered as an appropriate genetic vaccine candidate for H. pylori in future investigations.

Prevalence of &amp;lt;i&amp;gt;Helicobacter pylori vacA&amp;lt;/i&amp;gt;, &amp;lt;i&amp;gt;cagA&amp;lt;/i&amp;gt;, &amp;lt;i&amp;gt;dupA&amp;lt;/i&amp;gt; and &amp;lt;i&amp;gt;oipA&amp;lt;/i&amp;gt; Genotypes in Patients with Gastric Disease

Gastric diseases such as chronic gastritis and gastric cancer are most commonly caused by virulence factors of Helicobacter pylori (H. pylori), such as the vacA, cagA, dupA and oipA genes. Therefore, this study investigated the prevalence and the combination of these virulence factors from patients with gastric diseases. The endoscopic biopsies were obtained from 516 patients with gastric symptoms, 101 of which were from patients with normal gastric tissue, 365 of which were from patients with chronic gastritis, and 50 of which were from patients with gastric cancer. H. pylori and the virulence factors were detected by PCR. The oipA gene exhibited an increased risk for chronic gastritis (p = 0.0296), and the vacA gene demonstrated a risk for gastric cancer from chronic gastritis (p = 0.0002). Based on the combination of the virulence factors, cagA, vacA, dupA and oipA genes exhibited a high prevalence in patients with chronic gastritis and gastric cancer. The cagA+/dupA+ genotype demonstrated a significant correlation in patients with normal gastric mucosa (p = 0.0278). In the chronic gastritis group, a significant association was observed between the cagA+ and the vacA s1m1 genotypes (p cagA+/dupA+ genotypes (p = 0.0183), the dupA+/oipA+ genotypes (p cagA, vacA, dupA, and oipA genes, which contributed to the risk of developing gastroduodenal diseases. Furthermore, this is the first study to reveal a high prevalence of the oipA gene in H. pylori isolates in Brazil.

INVESTIGATION OF Helicobacter pylori VIRULENCE GENOTYPE IN GASTRIC BIOPSIES BY PCR

Background: Helicobacter pylori infections has been associated with the genetic diversity of their virulence factors, the virulence genotypes are valuable as molecular marker in the diagnosis of patients with bacterial infections . Our main objective was to analyze the frequency and allelic genotype of vacA , cagA also investigate another virulence genes of H. pylori. Methods: 75 biopsies of patients with gastritis and peptic ulcer diseases were selected to investigate the presences of H. pylori and collected from them antrum biopsies, then genomic DNA was extracted from antrum biopsies using genomic DNA kit .Subsequently, the virulence genes of H. pylori were amplified using specific primers including vacA , cagA, cagE and oipA and iceA by PCR in 49 cases that positive to 16SrRNA which previously investigated. Results: A high prevalence of genes cagA (28.6%), vacAs1bm2 (56.8%), iceA2 (30.6%) and oipA (42.9%) was found, while vacA s2m1 and iceA1 genotypes was not found in our study. There was significant correlation between the presence of cagA and cagE genotypes (p = 0.02), suggesting that these two genes almost used together as cag PAI integrity marker. The presence of cagA gene was significantly associated with peptic ulceration (p ≤ 0.001), whereas different vacA genotypes or iceA2 genotype were no statistically significant with clinical outcome. Patients with peptic ulcer disease more likely to have oipA gene (61.9% ) than those with gastritis (38.1%), P = 0.037, also the presence oipA gene was statistically significant with presence iceA2. Conclusion : Most H. pylori genotypes which associated with peptic ulcer and gastritis were moderate virulent strains.