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T Wadström

Publications and source records attributed to T Wadström.

At least 73 records · Page 4Linked to original sources
PubMed1998

Plasminogen binding and activation at the surface of Helicobacter pylori CCUG 17874.

The binding of iodine-labelled plasminogen to Helicobacter pylori CCUG 17874 was characterized. Inhibition of the binding was observed after preincubation of H. pylori cells with nonradiolabelled plasminogen, lysine, or the lysine analogue epsilon-aminocaproic acid. Fragments of plasminogen, kringles 1 to 3, kringle 4, and mini-plasminogen, were also studied as potential inhibitors. Mini-plasminogen caused total inhibition of the plasminogen binding, while the other fragments caused only partial inhibition. These findings suggest that H. pylori binds specifically the fifth kringle structure of the plasminogen molecule. Plasminogen binding to H. pylori seems to be independent of culture media and independent of the presence of the cytotoxin-associated CagA antigen. Immunoblot analysis identified two plasminogen binding proteins of 57 and 42 kDa. Scatchard plot analysis revealed one binding mechanism with a Kd value of 7 x 10(-7) M. Conversion of H. pylori cell-bound plasminogen to plasmin in the presence of a tissue-type plasminogen activator was demonstrated by digestion of the chromogenic substrate S-2251. No activation was noted when plasminogen or tissue-type plasminogen activator was incubated with H. pylori cells alone. Formation of H. pylori cell surface-bound plasmin may be important to provide a powerful proteolytic mechanism for gastric tissue penetration in type B gastritis and peptic ulcer disease, since plasmin degrades not only fibrin but also extracellular matrix proteins such as various collagens and fibronectin.

Aminocaproic Acid↗
PubMed1997

Binding of extracellular matrix proteins by enterococci.

Forty-four enterococcal strains isolated from human clinical specimens were investigated for binding of 125I-labeled fibronectin, vitronectin, thrombospondin, lactoferrin, and collagen type I and IV, and for cell surface hydrophobicity. Most strains expressed low binding of iodine-labeled human fibronectin, collagen I and IV, and higher binding of human vitronectin, human lactoferrin, and human thrombospondin. Bacteria grown in Todd-Hewitt broth exhibited increased binding to vitronectin and thrombospondin. In particle agglutination assays (PAA), Enterococcus faecalis strains reacted strongly with coated latex beads in contrast to E. faecium strains, which generally did not react. The ability of enterococci to bind ECM proteins was affected by heating and proteolytic digestion, suggesting that some protein-binding components become surface exposed after treatment with proteases. The binding of 125I-labeled proteins to E. faecalis strain E70 was inhibited when cells were preincubated with unlabeled proteins. Preincubating cells with sulfated polymers such as dextran sulfate (Mr 5000 and 8000), pentosan sulfate and heparin decreased binding of vitronectin, lactoferrin, and thrombospondin. The binding of lactoferrin and thrombospondin was also decreased when bacteria were preincubated with galactose, fucose, and mannosamine, but not with mannose. All of 30 E. faecalis strains expressed pronounced surface hydrophobicity, but 10 of 14 E. faecium strains showed hydrophilic cell surface.

Ammonium Sulfate↗
PubMed1997

Sulfatides inhibit binding of Helicobacter pylori to the gastric cancer Kato III cell line.

Helicobacter pylori adhere to Kato III and Hela S3 cells in monolayer cultures. To explore whether cell surface glycoconjugates on these two cell lines mediate binding of H. pylori, various carbohydrates, glycoproteins, and glycolipids were tested to inhibit H.pylori cell adhesion. The adhesion was measured (i) with a urease-based assay and (ii) by cells stained with fluorescein. Sodium periodate and sialidase treatment (but not alpha- or beta-galactosidase, heparitinase,lysozyme, or trypsin) inhibited H. pylori binding to both cell lines. Sulfatides and sulfated glycoconjugates (50 microg/ml) but not heparin or a number of simple carbohydrates inhibited binding (1 mg/ml). The two H.pylori strains studied (CCUG 17874 and strain 25) showed high binding of soluble 125I-labeled heparin and other sulfated carbohydrate compounds.

Bacterial Adhesion↗
PubMed1997

The Helicobacter pylori seroprevalence in blood donors related to Lewis (a,b) histo-blood group phenotype.

OBJECTIVE: To study a possible association of the Helicobacter pylori seroprevalence with ABO(H) and Lewis (a,b) blood group phenotypes in blood donors. DESIGN: A cross-sectional study of blood donors using ABO(H) and Lewis (a,b) blood group phenotype as predictors. METHODS: ABO(H) and Lewis (a,b) blood group phenotyping was performed with monoclonal antibody. The H. pylori immunoglobulin G (IgG) antibody relative activity was evaluated by enzyme-linked immunosorbent assay (ELISA) using acid glycine extract from H. pylori. SUBJECTS: One hundred and fifty-nine randomly selected blood transfusion donors. RESULTS: The individuals with Lewis (a+b-)/non-secretor phenotype showed a significantly higher proportion of the H. pylori-seronegative subjects and a lower IgG immune response to H. pylori antigens as compared with the individuals of Lewis (a-b+)/secretor phenotype. CONCLUSION: The Lewis (a,b) histo-blood group antigens are implicated in the mechanisms of naturally occurring resistance to H. pylori infection.

ABO Blood-Group System↗
PubMed1997

Helicobacter pylori-human polymorphonuclear leucocyte interaction in the presence of ammonia.

OBJECTIVE: To determine if the ammonia produced by Helicobacter pylori affects the phagocytic ability of human polymorphonuclear leucocytes as measured by the oxidative burst. METHODS: Interactions between opsonized urease-positive and -negative strains of H. pylori with polymorphonuclear leucocytes were studied in two series of experiments. In the first series of experiments, concentrations from 0 to 50 mM of NH4Cl were added to polymorphonuclear leucocytes. In the second series of experiments, bacteria were pre-incubated for 1 h with urea (0 to 50 mM) before addition of phagocytes. Luminol-dependent chemiluminescence was measured every 5 min over a 50-min period. The pH was verified in each treatment. RESULTS: Inhibition of chemiluminescence, increasing with concentration, was noted in all treatments when NH4Cl was added. When urea was added to urease-positive strains, chemiluminescence was significantly reduced when compared to the urease-negative strain and the zymosan control. This effect could not be attributed to a change in pH in the experiments using NH4Cl or urea at a concentration of 5 mM and 10 mM. CONCLUSION: Ammonia generated by H. pylori may contribute to the decreased activity of polymorphonuclear leucocytes in vivo.

Ammonia↗
PubMed1997

Identification of heparan sulphate binding surface proteins of Helicobacter pylori: inhibition of heparan sulphate binding with sulphated carbohydrate polymers.

Heparan sulphate binding to cells of the gastric pathogen Helicobacter pylori at pH 4-6 is common. Binding was inhibited by various unlabelled sulphated polysaccharides and at high ionic strength and pH, but not by carboxylated or non-sulphated compounds. The inhibition by various sulphated compounds such as dextran sulphate and carrageenans was related to the sulphate content and not to the carbohydrate polymer backbone. The IC50 values for heparin and dextran sulphate for H. pylori strain 25 were calculated as 3.55 x 10(-7) M and 5.01 x 10(-6) M respectively. Heparin-binding proteins of H. pylori are exposed on the cell surface, as shown by biotinylation of cell-surface proteins before separation of outer membranes and by an indirect immunofluorescence assay. The strongest biotin-heparin binding by H. pylori was observed with a polypeptide in the 55-60 kDa region.

Bacterial Outer Membrane Proteins↗
PubMed1997

Infection of BALB/c A mice by spiral and coccoid forms of Helicobacter pylori.

Helicobacter pylori exists in two different morphological forms, spiral and coccoid. This study demonstrated that both forms can infect BALB/c A mice. The animals were inoculated orally three times at 2-day intervals with 10(8) cfu of both spiral and coccoid forms of strain CCUG 17874 (NCTC 11637), strain 25 and strain 553/93. Infection was followed over a 30-week period by histological scoring of the grade of inflammation in gastric biopsies. At each time point sera were collected for analysis in ELISA and immunoblot analysis. Both spiral and coccoid forms of all H. pylori strains gave significantly higher inflammation scores than a control group of animals 1 week after inoculation. The histological evidence persisted throughout the entire 30 weeks. The inflammation was most severe in the pylorus and duodenum. Infection with strain 553/93 displayed the most severe gastritis. The spiral form of strain CCUG 17874 gave an immune response after only 4 weeks, whereas its coccoid form as well as strains 25 and 553/93 (spiral and coccoid forms) gave a significant increase in antibody response in ELISA and immunoblot after 16 weeks. It is concluded that both spiral and coccoid forms of H. pylori can cause acute gastritis in BALB/c A mice.

Animals↗
PubMed1997

Lack of evidence for sialidase activity in Helicobacter pylori.

The role of sialic acid for the adhesion of Helicobacter pylori to gastric mucosa cells and/or to the mucin layer is still under debate. Several but not all H. pylori strains express a sialic acid-binding adhesin, specific for terminal alpha-2,3-sialic acid residues. Recently, the production of sialidase by H. pylori was reported [Dwarakanath, A.D. et al. (1995) FEMS Immunol. Med. Microbiol. 12,213 216]. We analysed several strains isolated from gastric biopsies cultivated both in liquid media and on agar plates for sialidase. Activity of this enzyme was first assayed using the fluorigenic substrate 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid. Since the fluorimetric assay can give false-positive results caused by non-specific interactions with umbelliferyl-tagged substances, we used also the more sensitive and specific assay with sialyl-[3H]lactitol as a substrate. No evidence for sialidase activity of H. pylori strains, cultivated under both inducible and non-inducible conditions, was obtained.

Dyspepsia↗
PubMed1997

Identification of the N-acetylneuraminyllactose-specific laminin-binding protein of Helicobacter pylori.

The interaction of the gastroduodenal pathogen Helicobacter pylori with the glycoprotein laminin was investigated. Binding of 125I-radiolabelled laminin in a liquid-phase assay by both hemagglutinating and poorly hemagglutinating strains was rapid, saturable, specific, partially reversible, of high affinity, and insensitive to pH. Inhibition of laminin binding by fetuin, but not asialofetuin, and reduced bacterial binding to periodate- or sialidase-treated laminin indicated that glycosylation, particularly sialylation, was important for laminin binding by H. pylori. Inhibition experiments with monosaccharides, disaccharides, and trisaccharides showed that the strains bound to a region spanning a trisaccharide. In particular, inhibition and displacement studies showed that binding to the trisaccharide N-acetylneuraminyl-alpha(2-3)-lactose [NeuAc(2-3)Lac] was preferential to that to the NeuAc(2-6)Lac isomer. Complete inhibition of laminin binding by both hemagglutinating and poorly hemagglutinating strains was achieved only when isolated lipopolysaccharide (LPS) was used as an inhibitor in combination with heat or protease treatment of H. pylori cells, thereby confirming the involvement of both LPS and a protein adhesin in laminin binding. Further inhibition experiments indicated that the protein receptor, rather than LPS, on H. pylori bound NeuAc(2-3)Lac. By using a Western blotting procedure, a 25-kDa outer membrane protein was identified as mediating laminin binding by both hemagglutinating and poorly hemagglutinating H. pylori strains. The specificity of binding was confirmed by complete inhibition of laminin binding by the 25-kDa protein with NeuAc(2-3)Lac. The data collectively suggest that a 25-kDa outer membrane protein acts in a lectin-like manner with LPS to mediate attachment of H. pylori to laminin.

Adhesins, Bacterial↗
PubMed1997

Immunoblot assay for serodiagnosis of Helicobacter pylori infections.

An immunoblot assay for the serological diagnosis of Helicobacter pylori infection was evaluated. Serum samples from patients whose gastric biopsy specimens were known to be positive or negative for H. pylori on culture were used to establish interpretive criteria for the immunoblot assay. A panel of sera from patients with diseases other than H. pylori infection and sera from healthy blood donors were included to validate these criteria. All sera were initially assessed in an enzyme immunoassay (Ge-EIA), based on acid glycine-extracted cell surface proteins of H. pylori NCTC 11637. The same antigen extract was used in the immunoblot assay. In addition, the Ge-EIA and the immunoblot assay were compared with a commercially available EIA (Seradyn, Color Vue Pylori). Bands of 110/120 kDa and/or two of five low-molecular-mass proteins (26, 29, 30, 31, and 33 kDa, in any combination) showed a strong correlation with the H. pylori culture-positive patients (97.5%) compared to the correlation obtained with the EIA results (Ge-EIA, 87.5%; Seradyn EIA, 92.5%), and the antibody responses to these proteins were considered specific reactions. In 37 of 40 serum samples from culture-negative patients and also in sera from patients with other disorders, a moderate antibody reactivity to the medium-size proteins (43 to 66 kDa) was observed, and these were considered not valuable for a specific immunoblot assay. Among sera from culture-positive patients, 39 of 40 serum samples were defined to be immunoblot positive, and from among sera from culture-negative patients, 3 of 40 serum samples were defined to be immunoblot positive. The use of sera from patients with negative cultures for H. pylori as negative controls may decrease the sensitivity due to sampling error and false-negative culture results. Immunoblot assay-positive results were detected among 10% of sera from patients with other diseases, whereas they were detected among 42.5% of sera by the Ge-EIA and 47.5% of sera by the Seradyn-EIA. The higher number of EIA-positive sera in this group reflects a possible cross-reactivity (false-positive EIA result). Of the blood donors, representing asymptomatic but possibly colonized subjects, 24% were immunoblot positive. In conclusion, our data indicate that immunoblotting is more sensitive as well as more specific than EIA. Moreover, it permits detection of antibody responses to specific antigens, e.g., the cytotoxin-associated CagA protein, which may have pathological implications.

Adult↗
PubMed1997

Biochemical aspects of H. pylori adhesion.

In analogy with Vibrio cholerae and other toxigenic enteropathogens such as Yersinia enterocolitica it seems most likely today that H. pylori by specific surface proteins binds to epithelial cell receptors (Table 1) allowing the pathogen to deliver urease, vacuolating toxin and other toxic metabolites to cause tissue damage and inflammation (1-3). H. pylori as well as animal gastric pathogens as H. mustelae and H. felis have developed an efficient flagellar apparatus allowing rapid and efficient penetration of the gastric mucus layer (3). Most interestingly, flagellae seems to be synthesized in the late stage of growth before the switch of spiral-shaped organisms to coccoidal forms in a liquid medium.

Bacterial Adhesion↗
PubMed1997

Attachment of Helicobacter pylori strains to human epithelial cells.

The aim of the study was to characterize several clinical isolates of H. pylori as regards the activity and specificity of their haemagglutinins and the involvement of surface sialic acid-specific and heparin-binding compounds in the adhesin of the bacteria to human epithelial cell lines. Although H. pylori strains caused haemagglutination (HA) of sheep erythrocytes, they differed markedly by activity and specificity. On the basis of haemagglutination inhibition study three types of H. pylori strains could be distinguished. The HA of Type I strains was inhibited with fetuin/mucin but not asialofetuin/asialomucin. The HA activity of Type II strains was inhibited with fetuin/mucin and asialofetuin/asialomucin. The HA of Type III strains was not influenced by any of these inhibitors. In vitro, H. pylori strains bound to the cells of human epithelial lines: HeLa, Kato-3, Ags. However, various compounds mediated the binding of H. pylori types distinguished by HA, to epithelial cells. The interaction of some of H. pylori strains with epithelial cells was mediated by bacterial sialic acid-binding compounds. The majority of H. pylori strains used heparin-binding surface compounds to attach to epithelial cells. Clinical H. pylori strains differ by the compounds used in adhesin to epithelial cell lines, however, this process also depends on the expression of appropriate receptors on the host cells.

Bacterial Adhesion↗
PubMed1996

Immunochemical properties of a 60 kDa cell surface-associated heat shock-protein (Hsp60) from Helicobacter pylori.

Western blot analysis (immunoblotting) of cell surface-associated proteins from Helicobacter pylori confirmed our previous findings that binding of human IgG is a common property (among H. pylori strains). Purification of the IgG-binding proteins (IGBP) was achieved by two purification steps, affinity chromatography on IgG-Sepharose and nickel chelate affinity chromatography. SDS-PAGE and immunoblotting analysis revealed a 60 kDa protein with affinity for peroxidase labeled human IgG. Solid phase binding assays showed that IgG binds to an immobilized protein (IGBP). The 60 kDa IGBP binds human IgG1, IgG3 and IgM. Binding could be inhibited by the kappa chain of the human IgG, but not with its Fc fragment, nor with IgA or IgM. In addition, rabbit polyclonal antibodies raised against the 60 kDa IGBP blocked IgG binding. Monoclonal antibodies, specific to the Hsp60 heat shock protein of H. pylori recognized the 60 kDa IGBP as revealed by immunoblotting analysis, both in crude preparations and in the purified fractions.

Antibodies, Bacterial↗
PubMed1996

Cryptic domains of a 60 kDa heat shock protein of Helicobacter pylori bound to bovine lactoferrin.

Bovine lactoferrin binds to a 60 kDa heat shock protein of Helicobacter pylori. Binding ability was related to human immunoglobulin G because bovine lactoferrin binding proteins were isolated by extraction of cell surface associated proteins with distilled water, applied on IgG-Sepharose and nickel sulphate chelate affinity chromatography. Binding was demonstrated by Western blot after purified protein was digested with alpha-chymotrypsin and incubated with peroxidase-labeled bovine lactoferrin. Binding was inhibited by bovine lactoferrin, lactose, rhamnose, galactose, and two iron-containing proteins, ferritin and haptoglobin. Helicobacter pylori binds ferritin and haptoglobin via charge or hydrophobic interactions because this binding was not inhibited by specific and various glycoproteins or carbohydrates. Carbohydrate moieties of bovine lactoferrin molecules seem to be involved in binding because glycoproteins with similar carbohydrate structures strongly inhibited binding. Scatchard plot analysis of the binding of peroxidase-labeled bovine lactoferrin to H. pylori cells yielded a kd 2.88 x 10(-6) M. In addition, binding of H. pylori cells to bovine lactoferrin was enhanced when bacteria treated with pepsin or alpha-chymotrypsin after isolation from iron-restricted and iron-containing media.

Animals↗
PubMed1996

A collagen binding protein from Lactobacillus reuteri is part of an ABC transporter system?

The gene coding for a collagen binding protein from Lactobacillus reuteri NCIB 11951 was cloned and sequenced. A genomic lambda library was constructed and recombinant plaques were screened using antisera raised against purified collagen binding proteins from the same L. reuteri strain. The positive plaques were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis, which revealed the expression of a 29 kDa protein, which reacted with the antisera and bound 125I-labelled type I collagen. The sequence of the corresponding gene, cnb showed that the collagen binding protein has sequence similarities to the solute binding component of bacterial ABC transporters.

Amino Acid Sequence↗
PubMed1996

IgG immune response to Helicobacter pylori antigens in patients with gastric cancer as defined by ELISA and immunoblotting.

Helicobacter pylori infection is considered to be a risk factor for gastric cancer. A high prevalence of H. pylori infection and high gastric-cancer incidence are characteristic of the Estonian population. To evaluate the relationship between these 2 events, we studied the seroprevalence of H. pylori in gastric cancer patients (n = 182) and in healthy blood donors (n = 306). A relative anti-H. pylori IgG antibody activity, as detected by ELISA and immunoblot patterns, was correlated with age, stage of the disease and tumor morphology. A significantly higher H. pylori seroprevalence was found in patients in the early stages of tumor development compared with both advanced cancer patients and controls. No significant difference in H. pylori seroprevalence between patients with the intestinal and diffuse types of tumor growth was observed. A decline in the recognition of putatively cross-reacting (33-66 kDa) antigens was noted in the cancer group. The response to vacuolating toxin-related 85-kDa and CagA 120-kDa protein antigens was not altered and was observed more often in the younger group of cancer patients.

Adult↗