Construction of expression systems forflaAandflaBgenes ofHelicobacter pyloriand determination of immunoreactivity and antigenicity of recombinant proteins

Yan, Jie; Liang, Shuli; Mao, Yafei; Li, Liwei; Li, Shuping

doi:10.3748/wjg.v9.i10.2240

Cited by 16 publications

(22 citation statements)

References 42 publications

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“…It was reported that there are many protective antigens against H pylori, such as UreB, HpaA, VacA, CagA, NapA, FlaA, FlaB and heat shock protein (HSP) [8][9][10][11][12][13]23] . Due to cross-antigens against HSP in different sibling bacteria [23] , 7 kinds of protective antigens of H pylori except for HSP were selected in our research.…”

Section: Discussionmentioning

confidence: 99%

“…So far, the gene orders of ureB, hpaA, napA, flaA and flaB have been confirmed as highly conservative, while the gene order of vacA can mutate moderately between signal peptide zone (S) and intermedial zone (M), and the mutations of CagA gene order are mainly located at the 3' terminal repeat [8,9,12,13,15] . Our research focused on the carrying rates and expression level of the 7 kinds of genes of H pylori from the clinical H pylori in China.…”

Section: Discussionmentioning

confidence: 99%

“…For example, HpaA is usually called an adhesin of H pylori, and in fact, it is a tunica vaginalis protein expressed in the flagellae of H pylori [9] . FlaA and FlaB expressed in the flagella are subunits of flagellin [13] . As an intein of H pylori, CagA is a non-exocrine protein and only when H pylori has contact with host cells, does it act on the target cells through type Ⅲ secretory system [10,11] .…”

Section: Discussionmentioning

confidence: 99%

“…Both the upstream and downstream primer concentrations were 20 pmol. Primer sequences, main reaction values and the product are shown in Table 1 [13,[18][19][20][21][22] . The PCR products were identified with 2% agarose gel staining using the fluorescent dye ethidium bromide.…”

Section: Detection Of Target Genesmentioning

confidence: 99%

“…The selection for antigenic targets is critical in the design of a genetically engineered vaccine against H pylori. The best ascertained antigens out of the outer membrane protein antigens are UreB and HpaA followed by VacA, CagA, NapA, FlaA and FlaB [8][9][10][11][12][13] . Tomb reported all the genomic sequences of H pylori in 1997 [14] , and in our previous study, these genomic prokaryotic expression systems were constructed.…”

Section: Introductionmentioning

confidence: 99%

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Diversity of Helicobacter pylori isolates in expression of antigens and induction of antibodies

Tang¹,

Luo²,

Sun³

et al. 2008

WJG

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AIM:To obtain evidence for selection of antigens used in genetically engineered vaccine against Helicobacter pylori (H pylori ). METHODS:Enzyme linked immunoabsorbent assay (ELISA) was established on the basis of recombinant protein antigens rUreB, rHpaA, rVacA, rCagA1, rNapA, rFlaA and rFlaB of H pylori to detect expression rates of the antigens in bacterial isolates as well as positive rates of the antibodies in sera from H pylori -infected patients. PCR was applied to the detection of carrying rates of the genes encoding antigens in the isolates. RESULTS: The outputs of rUreB, rHpaA, rVacA, rCagA1, rNapA, rFlaA and rFlaB were approximately 35%, 32%, 15%, 23%, 56%, 25% and 20% of the total bacterial proteins, respectively. One hundred and fifty-one strains of H pylori were isolated from 347 biopsy specimens of chronic gastritis, peptic ulcer or gastric adenocarcinoma, with a positive rate of 43.5%. All of the isolates expressed UreB, HpaA, FlaA and FlaB while 52.3%, 92.1% and 93.4% of the isolates expressed VacA, CagA and NapA, respectively. In the sera of 151 H pylori -infected patients, the positive rates of IgG antibodies against UreB, HpaA, VacA, CagA, NapA, FlaA and FlaB were 100%, 87.4%, 43%, 71.5%, 89.4%, 84.8% and 79.5%, respectively. Furthermore, the expression frequencies of VacA and NapA were found to be relative to the severity of gastric diseases (P = 0.016 and P < 0.0001, respectively). CONCLUSION:UreB antigen is the top option of developing genetically engineered vaccine against H pylori followed by NapA or HpaA.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Detection Of Target Genesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Diversity of Helicobacter pylori isolates in expression of antigens and induction of antibodies

Tang¹,

Luo²,

Sun³

et al. 2008

WJG

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Potential antigen candidates for subunit vaccine development against Helicobacter pylori infection

Eslami

Yousefi

Ghasemian

et al. 2019

Journal Cellular Physiology

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Helicobacter pylori (H. pylori) is a resident bacterium in the stomach that accounts for 75% cases of gastric cancer. In this review, we comprehensively studied published papers on H. pylori vaccines using Google Scholar and NCBI databases to gather information about vaccines against H. pylori. Considering the pivotal roles of the enzyme urease (in production of NH 3 and neutralization of the acidic medium of the stomach), cytotoxinassociated gene A, and vacuolating cytotoxin A proteins in H. pylori infection, they could be the best candidates for the construction of recombinant vaccines. The outer membrane porins (Hop), blood group antigen-binding adhesin (BabA), sialic acid-binding adhesin (SabA), and outer inflammatory protein A, play significant roles in binding of bacterium to human gastric tissues, and because binding is the first step in bacterial fixation and colonization, these antigens also can be considered as suitable candidates for designing vaccines. Likely, other significant bacterial antigens, such as NapA (chemotactic factor for recruitment of human neutrophils and monocytes to the site of infection), duodenal ulcer promoting protein A (to promote duodenal ulcer), and Hsp60 (as a molecular chaperon for activation of urease enzyme), can be used in the construction of subunit vaccines. New vaccines in use currently, such as DNA vaccines and subunit vaccines, can efficiently replace the dead and attenuated vaccines. Nonetheless, the results show that urease enzyme is most used compared with bacterial components in the designing and construction of recombinant vaccines. The BabA and SabA antigens belong to the outer membrane porins family in H. pylori and are required for binding and fixation of the bacterium to the human gastric tissues. K E Y W O R D SCagA, Helicobacter pylori, recombinant vaccine, urease, VacA

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Antigenic and conserved peptides from diverse Helicobacter pylori antigens

Calado

2022

Biotechnol Lett

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Since the revolutionary finding of Helicobacter pylori as a common bacterial infection, that a high research effort for its eradication has been conducted. Epitope based-vaccine presents advantages over protein-based, as they can be designed to contain epitopes from diverse proteins, therefore, more easily representing the immune-variability of the bacterial population, while minimizing the toxicity associated to some whole proteins. In the present work, an iterative method, to design antigenic and conserved B-epitopes from diverse virulent factors of H. pylori, was established. The method considered the trade-off between epitopes antigenicity and conservation among the bacterial population. For the method validation, five virulent factors from H. pylori were selected. From each virulent factor, two epitopes were predicted, each with twelve residues of aminoacids. The corresponding ten peptides were synthesised and evaluated by enzyme-linked immunosorbent assay using polyclonal antibodies raised against a specific H. pylori strain. All ten peptides were recognised by the antibodies and were consequently antigenic and conserved. This result could strongly contribute to the design of a multivalent epitope-based vaccine, representing the immunogenetic variability within the bacterial population, leading to a sustained and effective immunogenic protection.

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Construction of expression systems forflaAandflaBgenes ofHelicobacter pyloriand determination of immunoreactivity and antigenicity of recombinant proteins

Cited by 16 publications

References 42 publications

Diversity of Helicobacter pylori isolates in expression of antigens and induction of antibodies

Diversity of Helicobacter pylori isolates in expression of antigens and induction of antibodies

Potential antigen candidates for subunit vaccine development against Helicobacter pylori infection

Antigenic and conserved peptides from diverse Helicobacter pylori antigens

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