Recombinant Mycobacterium smegmatis mc2155 vaccine expressing outer membrane protein 26kDa antigen affords therapeutic protection against Helicobacter pylori infection

ABSTRACTHelicobacter pyloriinfection is prevalent worldwide and results in chronic gastritis, which may lead to gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer. We have previously reported that oral immunization with recombinantMycobacterium smegmatisexpressing theH. pyloriouter membrane protein 26-kilodalton (Omp26) antigen affords therapeutic protection againstH. pyloriinfection in mice. In the present study, we investigated the prophylactic effects of this vaccine candidate onH. pylorichallenge in mice. We found that oral immunization with recombinantMycobacteriumOmp26 significantly reducedH. pyloricolonization in the stomach compared to inoculation with wild-typeM. smegmatisin control mice. Six of the recombinantMycobacterium-immunized mice (60%) were completely protected fromH. pyloriinfection. The severity ofH. pylori-associated chronic gastritis assessed histologically was significantly milder in mice vaccinated with recombinantMycobacteriumthan in control animals. Mice immunized with recombinantMycobacteriumshowed enhanced antigen-specific lymphocyte proliferation and antibody responses. Moreover, immunization with recombinantMycobacteriumresulted in an increased expression of interleukin-2 and gamma interferon in the stomach and spleen, as determined by reverse transcription-PCR analysis. Our results collectively suggest that vaccination with recombinantMycobacteriumOmp26 confers prophylactic protection againstH. pyloriinfection. The inhibition ofH. pyloricolonization is associated with the induction of antigen-specific humoral and cell-mediated immune responses.

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Faculty Opinions recommendation of Helicobacter pylori outer membrane protein HopQ identified as a novel T4SS-associated virulence factor.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718021621.793488300 ◽

2013 ◽

Author(s):

Timothy Cover

Keyword(s):

Helicobacter Pylori ◽

Membrane Protein ◽

Outer Membrane ◽

Virulence Factor ◽

Outer Membrane Protein

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The Helicobacter pylori Autotransporter ImaA (HP0289) Modulates the Immune Response and Contributes to Host Colonization

Infection and Immunity ◽

10.1128/iai.00312-12 ◽

2012 ◽

Vol 80 (7) ◽

pp. 2286-2296 ◽

Cited By ~ 13

Author(s):

William E. Sause ◽

Andrea R. Castillo ◽

Karen M. Ottemann

Keyword(s):

Immune Response ◽

Helicobacter Pylori ◽

Membrane Protein ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Dependent Manner ◽

Wild Type ◽

Content Type ◽

H Pylori ◽

Murine Infections

ABSTRACTThe human pathogenHelicobacter pyloriemploys 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 geneHP0289, which encodes an uncharacterized outer membrane protein. We first generated an isogenicH. pylorimutant that lacksHP0289and found that the mutant has a colonization defect in single-strain infections and is greatly outcompeted in mouse coinfection experiments with wild-typeH. 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 theHP0289promoter is upregulated in the mouse stomach, and here we demonstrate thatHP0289expression is induced under acidic conditions in an ArsRS-dependent manner. Finally, we have shown that theHP0289mutant 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-typeH. pylori. On the basis of this phenotype, we renamed HP0289 ImaA forimmunomodulatoryautotransporter protein. Our work has revealed that genes inducedin vivoplay an important role inH. pyloripathogenesis. Specifically, the outer membrane protein ImaA modulates a component of the host inflammatory response, and thus may allowH. pylorito fine tune the host immune response based on ImaA expression.

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Helicobacter pylori outer membrane protein, HomC, shows geographic dependent polymorphism that is influenced by the Bab family

The Journal of Microbiology ◽

10.1007/s12275-016-6434-8 ◽

2016 ◽

Vol 54 (12) ◽

pp. 846-852 ◽

Cited By ~ 3

Author(s):

Aeryun Kim ◽

Stephanie L. Servetas ◽

Jieun Kang ◽

Jinmoon Kim ◽

Sungil Jang ◽

...

Keyword(s):

Helicobacter Pylori ◽

Membrane Protein ◽

Outer Membrane ◽

Outer Membrane Protein

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Identification of Functional Interactome of Gastric Cancer Cells with Helicobacter pylori Outer Membrane Protein HpaA by HPLC-MS/MS

BioMed Research International ◽

10.1155/2020/1052926 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Ruyue Fan ◽

Xiurui Han ◽

Di Xiao ◽

Lihua He ◽

Yanan Gong ◽

...

Keyword(s):

Gastric Cancer ◽

Helicobacter Pylori ◽

Epithelial Cells ◽

Membrane Protein ◽

Outer Membrane ◽

Cell Lines ◽

Outer Membrane Protein ◽

Regulation Of Transcription ◽

Gastric Epithelial Cells ◽

H Pylori

HpaA as an outer membrane protein of Helicobacter pylori (H. pylori) plays a significant role in the adhesion to the human stomach, but the functional relation between HpaA and gastric epithelial cells is still not clear. To screen the interaction between HpaA and cellular proteins in gastric epithelial cells, the HpaA protein from H. pylori 26695 fused with a tag (6× His) was expressed and purified successfully, the secondary structure was estimated by the Circular Dichroism (CD) spectrum, and the purified recombinant protein was used to perform the pull-down assays with gastric cancer cell lines (AGS and SGC-7901) lysates, respectively. The pull-down proteins were identified by high-performance liquid chromatography tandem mass spectrometry system (HPLC-MS/MS). A total of 9 and 13 proteins related were analyzed from AGS and SGC-7901 cell lysates, respectively. ANXA2 was considered as putative HpaA functional partner discovered from lysates of both cell lines with high score and coverage. It is hypothesized that HpaA may be involved in the biological process of regulation of transcription and nucleic acid metabolism during the adhesion of H. pylori to human gastric epithelial cells, and HpaA-binding proteins also be used as targets for the development of antiadhesion drugs against H. pylori.

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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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