Identification of erythrocyte-binding antigens in Helicobacter pylori (original) (raw)
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FEMS Immunology & Medical Microbiology, 1995
Rabbits were immunised with stage 1 and stage 2 soluble haemagglutinins @-IA) of Helicobacter pylori strain NCTC 11637 and with rabbit erythrocytes coated with stage 1 sH.A. After adsorption of stage 1 sHA on erythrocytes, SDS-PAGE analysis showed that 4 major protein bands were removed from the preparation. The anti-&% coated erythrocyte serum had the highest HA inhibition titre of 16. Crossed immunoelectrophoresis of the stage 1 &IA, against stage 1 and 2 antisera showed multiple precipitin arcs; however, the anti-&A coated erythrocyte serum produced only two arcs. One arc produced by the anti-stage 2 serum was absent with the anti-stage 1 serum. This arc could have been produced against a 20 kDa polypeptide which was albsent in the stage 1 sHA. The other arc was stronger when compared with that produced by anti-stage 1 serum. These two arcs corresponded to the two arcs produced by the anti-sHA coated erythrocyte serum, which had the highest inhibition titre. The two arcs were markedly reduced in crossed immunoelectrophoresis with an adsorbed stage 1 sHA preparation, which indicates that these arcs were produced against the sHAs.
Isolation of a sialic acid-specific surface haemagglutinin of Helicobacter pylori strain NCTC 11637
Zentralblatt für Bakteriologie, 1993
Untersuchung der mit Gold-markiertem Fetuin inkubierten H. pylori zeigte, dag das Hamagglutinin mit lose anhangendem Material an der Bakterienoberflache assoziiert war und dag das gereinigte Hamagglutinin keine Fimbrienstruktur aufwies. Die Fahigkeit zur Bindung an Sialoglycokonjugate der Erythrozytenmembran weist darauf hin, dag das Hammaglutinin einen wichtigen Besiedlungsfaktor darstellt, der H. pylori eine Bindung an ahnliche Saccharidstrukturen auf Epithelzellen ermoglicht.
IOSR Journal of Pharmacy and Biological Sciences, 2014
Helicobacter pylori (H.pylori) is a gram-negative microaerophilic bacterium that is generally associated with the main cause of approximately 80% peptic ulcers in stomach. It is also linked to the development of the stomach cancer. The aim of this study was to characterize the sub-unit pili proteins of Helicobacter pylori with a molecular weight of about 49,6 kDa as a haemagglutinin protein and adhesion molecules on surface of mice gastric epithelial cells. The bacterium was firstly cultured on the plate of TSA-B (Trypticase Soy Agar with 5% Sheep Blood) to prepare the protein of interes using bacterial cutter. The isolated proteins were subsequently analyzed for haemagglutination capability using mice sentisized erithrocytes. SDS-PAGE and western blotting analyses were used to confirm the protein patterns and their immunological reactivities respectively. The both SDS-PAGE and western blotting analyses showed that a dominant protein with molecular weight of about 49,6 kDa was detected consistently and found to be potentially aglutinate the erythrocytes. Furthermore, this protein was observed to be adherence on mice gastric epithelial cells. This study has indicated that the sub-unit pili proteins of H. pylori with a molecular weight of about 49,6 kDa was found to be dominant and immunogenic. Further study is required to confirm the in vivo properties of the protein associated with pathogenesis of H. pylori infection.
FEMS Microbiology Letters, 2000
We describe a new procedure allowing the generation and detection of immunogenic antigens from Helicobacter pylori via the hemolysin secretion apparatus of Escherichia coli. The gene (or gene fragment) encoding the H. pylori protein (or protein domain) is inserted in-frame into a residual portion of the hemolysin gene (hlyA), encoding the HlyA secretion signal (HlyA s ). These fusion proteins are secreted efficiently by E. coli. This new approach allows the identification of immunodominant antigens by using sera derived from H. pylori-infected patients suffering from different gastroduodenal pathologies. Three immunodominant antigens bearing the ureB (urease B-subunit), flaA (flagellin A-subunit), and an unknown ORF (HP0888) encoding an E. coli FecE analogous protein fused to hlyA s were identified and characterized. ß : S 0 3 7 8 -1 0 9 7 ( 0 0 ) 0 0 1 5 7 -9
Antigenic proteins of Helicobacter pylori of potential diagnostic value
Asian Pacific journal of cancer prevention : APJCP, 2013
Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (≥ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture- positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by m...
The neutrophil-activating protein of Helicobacter pylori
International Journal of Medical Microbiology, 2001
Infection of the stomach mucosa by the gastric pathogen Helicobacter pylori is accompanied by a large infiltration of neutrophils and monocytes which are believed to contribute substantially to H. pylori-induced gastritis. A protein was identified (HP-NAP for neutrophil-activating protein from H. pylori) that was capable of increasing the adhesion of neutrophils to endothelial cells. We have demonstrated that HP-NAP is a dodecamer composed of identical 17-kDa subunits that induces the production of reactive oxygen radicals (ROIs) by neutrophils via a cascade of intracellular activation events. HP-NAP has also been shown to be chemotactic for neutrophils and monocytes, and a majority of H. pylori-infected patients have been found to produce antibodies specific for HP-NAP making it a strong vaccine candidate. More recently it has been shown that HP-NAP can stimulate tissue factor and plasminogen activator inhibitor-2 production by human monocytes. While structurally similar to the Escherichia coli DNA-binding protein Dps, HP-NAP has characteristics that are more similar to bacterioferritins being capable of binding up to 500 atoms of iron in vitro. Further study, however, has revealed that synthesis of HP-NAP in H. pylori is not altered by the addition or subtraction of metal ions from its growth medium suggesting that the primary role of the protein in vivo is not as a metal-binding protein. A number of other reports have proposed that HP-NAP acts as an adhesin being capable of binding several different compounds in vitro. Sequence analysis of the genomes of several other bacteria reveal that many possess Dps/HP-NAP-like proteins. The preliminary characterisation of some of these proteins will be discussed.
Helicobacter pylori adhesins: HpaA a potential antigen in experimental vaccines for H. pylori
Helicobacter, 2020
Helicobacter pylori (H. pylori) is a gram-negative, microaerophilic bacteria containing between 2 and 6 unipolar flagella. The bacterium has rounded ends and measures 2.5-4 µm in length and 0.5-1 µm width. H. pylori selectively colonize the surface of the gastric mucosa of humans. 1-3 Since the discovery of H. pylori by Marshall and Warren, 4 numerous studies have demonstrated that this human gastric pathogen is present in approximately 50% of the world's population, and up to 55.8% in the Chinese population. 5,6 Until now, humans stomach have been recognized as the main reservoir for this pathogen. 7 Also, H. pylori have been isolated from nonhuman primates and domestic cats. 8-10 Sero-epidemiological surveys carried out on H. pylori reveal that the frequency of infection changes regionally, and it is lower in developed countries (30%-40%) than in developing countries (in some regions, >85%). 11,12 Most individuals remain asymptomatic. 13 However, long-term carriage of H. pylori considerably increase the risk of developing gastric diseases, such as gastritis (20%), peptic ulceration (10%), gastric adenocarcinoma (2%), and gastric mucosaassociated lymphoid tissue (MALT) lymphoma is less than 0.1% of those infected. 14,15 The routes of transmission of H. pylori are not yet fully understood. However, most infections are thought to be contracted in childhood via the oral-oral or fecal-oral routes. 16-19 H. pylori have been classified as a class I carcinogen in humans by a working group of the International Agency for Research on Cancer (IARC) of the World Health Organization(WHO). 20 So far, to fight H.
Production of a monoclonal antibody against the 128 kDa (CagA) protein of Helicobacter pylori
Journal of Immunological Methods, 1996
Helicobacrer pylon' is recognised as an important factor in gastroduodenal pathology. The 128 kDa CagA protein has been established as a useful marker of H. pyfori strains associated with more severe forms of disease. A mouse monoclonal antibody raised against the CagA protein has been produced and characterised as belonging to the IgGl subtype. It identified the protein in all clinical isolates (lo/IO) from this laboratory and in two NCTC reference strains (NCTC 11637 and NCTC 11961). No cross-reacting proteins were detected in H. pylori L2, a well characterised strain known not to contain the cugA gene, or in four Helicobacter sp. from non-human sources (H. cunis, H. mustelidae. H. muridarum and H. a&onyx). The monoclonal antibody was used to develop an antigen capture ELISA system for detecting the presence of antibodies to the CagA protein in human serum samples.
Separation and surveys of proteins of Helicobacter pylori
Journal of Chromatography B, 2002
The analysis of Helicobacter pylori proteins is a demanding task for the elucidation of virulence factors, antigens and vaccines, all important for diagnosis, therapy and protection. In the ''pre-genomic era'' the purification of proteins was mostly performed by using various techniques such as detergent treatment of the bacterial cells, ultra-centrifugation, various chromatographic methods, antibody detection, N-terminal sequence determination and finally cloning and identification of the corresponding gene. In this review, the most representative methods used for purification, separation and identification of H. pylori proteins will be presented as well as some important developments in the ''post-genomic era'' that have improved the performance of these characterisation techniques.