Immunogenic proteins ofHelicobacter pullorum,Helicobacter bilisandHelicobacter hepaticusidentified by two-dimensional gel electrophoresis and immunoblotting

Kornilovska, Iryna; Nilsson, Ingrid; Utt, Meeme; Ljungh, Åsa; Wadström, Torkel

doi:10.1002/1615-9861(200206)2:6<775::aid-prot775>3.0.co;2-r

Cited by 29 publications

(25 citation statements)

References 24 publications

(12 reference statements)

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“…Then, 10 l of serum was added to 1 ml of cell lysate (with the density adjusted at A 540 to 1.5), followed by incubation for 2.5 h at 22°C under constant shaking. Cells were removed by centrifugation at 12,000 ϫ g for 15 min, and the supernatants were used for the serology (21,49).…”

Section: Methodsmentioning

confidence: 99%

Cross-Reactivity between Immune Responses to Helicobacter bilis and Helicobacter pylori in a Population in Thailand at High Risk of Developing Cholangiocarcinoma

Pisani

Whary

Nilsson

et al. 2008

Clin Vaccine Immunol

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Helicobacter bilis DNA has been detected in human tissue and is a candidate for etiologic investigations on the causes of hepatic and biliary tract diseases, but reliable serologic tests need to be developed in order to pursue such investigations. The scope of this study was to assess the specificity of two assays for H. bilis immune response allowing for H. pylori, and their cross-reactivity in a population in Thailand at high risk for cholangiocarcinoma. Plasma samples from 92 Thai volunteers were independently tested in two laboratories (Massachusetts Institute of Technology [MIT] and Lund). MIT performed three analyses of H. pylori and H. bilis based either on (i) outer

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Section: Methodsmentioning

confidence: 99%

Cross-Reactivity between Immune Responses to Helicobacter bilis and Helicobacter pylori in a Population in Thailand at High Risk of Developing Cholangiocarcinoma

Pisani

Whary

Nilsson

et al. 2008

Clin Vaccine Immunol

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“…T and H. hepaticus strain CCUG 33637 T (murine isolates) were cultured on Brucella blood agar as described previously (Kornilovs'ka et al, 2002) for 4 days at 37 8C in a microaerobic atmosphere (Anoxomat; MART Microbiology). Helicobacter pylori strain CCUG 17874 T was cultured (low passage; fewer than four times) on GAB-CAMP agar (Soltesz et al, 1992) without antibiotics for 3 days at 37 8C in a microaerobic atmosphere (Anoxomat; MART Microbiology).…”

Section: Methodsmentioning

confidence: 99%

Increased prevalence of seropositivity for non-gastric Helicobacter species in patients with autoimmune liver disease

Nilsson

Kornilovska

Lindgren

et al. 2003

Journal of Medical Microbiology

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Various Helicobacter species have been isolated from the stomach, intestinal tract and liver of a variety of mammalian and some avian species, and Helicobacter DNA has been detected in human bile and liver samples. An immunoblot assay was established to analyse serum antibody responses to non-gastric Helicobacter species in patients with autoimmune liver diseases, in comparison with healthy individuals. Sera from 36 patients with primary sclerosing cholangitis (PSC), 21 with primary biliary cirrhosis, 19 with autoimmune chronic hepatitis and 80 blood donors were analysed by immunoblot, using cell-surface proteins from Helicobacter pullorum, Helicobacter bilis and Helicobacter hepaticus as antigens. Prior to testing, sera were cross-absorbed with a whole-cell lysate of Helicobacter pylori. Antibody reactivity to various proteins of these three Helicobacter species was measured by densitometric scanning and results were processed by computer software to estimate antigenic specificity. Results were also compared with antibody response to H. pylori. For H. pullorum, reactivity to at least two of the proteins with molecular masses of 48, 45, 37, 20 and 16 kDa, for H. hepaticus, reactivity to the 76, 30 and 21 kDa proteins and for H. bilis, reactivity to the 22 and 20 kDa proteins, seemed to have high specificity. Positive immunoblot results with sera from patients with PSC to antigens of H. pullorum, H. bilis and H. hepaticus were found in 38, 22 and 25 % of cases, respectively, and from patients with other autoimmune liver diseases, in 30, 22 and 22 % of cases, respectively. Prevalence of serum antibodies to non-gastric Helicobacter species was significantly higher in patients with autoimmune chronic liver diseases than in healthy blood donors (P , 0·001). Increased antibody levels to enterohepatic Helicobacter species raise questions concerning an infectious role of these emerging bacterial pathogens in human autoimmune liver diseases. INTRODUCTIONGenetic, environmental and microbial factors are believed to play a role in the development of chronic liver diseases of presumed autoimmune aetiology, such as primary sclerosing cholangitis (PSC) and primary biliary cirrhosis (PBC). Unknown environmental factors or infectious agents could trigger an immunological reaction and an inflammatory response that result in the destruction of intra-and extrahepatic bile ducts (Lee & Kaplan, 1995;Sutton & Neuberger, 2002). No causative microbial agent has been defined clearly for humans. Normal intestinal microflora or previously unrecognized intestinal pathogens might contribute to the development of disease in susceptible hosts. More recently, DNA of different Helicobacter species was detected in human bile and gall-bladder samples, suggesting that these organisms may colonize or translocate to the biliary tract of humans and may be associated with hepatobiliary diseases (Fox et al., 1998(Fox et al., , 2001Leong & Sung, 2002; Ljungh & Wadström, 2002).Isolation of a Helicobacter species from a human liver was reported recently (...

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“…In some experiments 2% serum was used and in others 5% was used, as indicated in the text or figure legends. All of the growth and amino acid uptake experiments were performed with 5% serum initially, since that level was used in previous H hepaticus studies (11,18). After it was determined that use of a lower serum level (2%) facilitated H 2 -dependent studies (see below), all (growth and amino acid transport) experiments were repeated with that serum level as shown.…”

Section: Bacterial Strains and Growth Conditions H Hepaticus Strainmentioning

confidence: 99%

Helicobacter hepaticus Hydrogenase Mutants Are Deficient in Hydrogen-Supported Amino Acid Uptake and in Causing Liver Lesions in A/J Mice

Mehta¹,

Benoit²,

Mysore

et al. 2005

Infect Immun

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Helicobacter hepaticus, a causative agent of chronic hepatitis and hepatocellular carcinoma in mice, expresses a nickel-containing hydrogen-oxidizing hydrogenase enzyme. Growth of a hyaB gene-targeted mutant was unaffected by the presence of hydrogen, unlike the wild-type strain, which showed an enhanced growth rate when supplied with H 2 . Hydrogenase activities in H. hepaticus were constitutive and not dependent on the inclusion of H 2 during growth. Addition of nickel during growth significantly stimulated both urease (for wild-type and hyaB) and hydrogenase (for wild-type) activities. In a 5-h period, the extent of 14 C-labeled amino acid uptake by the wild type was markedly enhanced in the presence of hydrogen and was >5-fold greater than that of the hyaB mutant strain. In the presence of H 2 , the short-term whole-cell amino acid uptake V max of the parent strain was about 2.2-fold greater than for the mutant, but the half-saturation affinity for amino acid transport was the same for the parent and mutant strain. The liver-and cecum-colonizing abilities of the strains was estimated by real-time PCR quantitation of the H. hepaticus-specific cytolethal distending toxin gene and showed similar animal colonization for the hyaB mutant and the wild type. However, at 21 weeks postinoculation, the livers from mice inoculated with wild type exhibited moderate lobular lymphoplasmacytic hepatitis with hepatocytic coagulative necrosis, but the hydrogenase mutants exhibited no histological evidence of lobular inflammation or necrosis.In recent years, more than three different Helicobacter species have been recovered from rodents (24,25,31), with H. hepaticus being the most well-studied enterohepatic Helicobacter species. First isolated in 1992 from untreated A/JCr control mice, H. hepaticus is a gram-negative, microaerophilic bacterium (6) that occurs naturally in many strains of inbred mice (30). Mice infected with this bacterium develop chronic hepatic lesions and are more prone to developing hepatocellular carcinoma (25, 26), which has made H. hepaticus an excellent model for studying mechanisms of bacterium-associated liver carcinogenesis. Although H. hepaticus can be isolated from the liver of infected mice, it is more consistently recovered from the intestinal tract since the primary site of colonization is the lower bowels of mice (6, 7). Recently, Helicobacter species DNA was reported to be associated with primary hepatocellular carcinomas in human liver samples (9, 23).At the same time, a hydrogen uptake hydrogenase enzyme well studied for its roles in nonpathogenic bacteria was demonstrated to be important for colonization of animals by some human pathogens (i.e., by Salmonella enterica serovar Typhimurium and Helicobacter pylori [14,21]); this sparked our interest in studying the physiological role of this enzyme in H. hepaticus (16). The colonization deficiency of hydrogenase structural gene mutants of H. pylori was attributed to their inability to utilize hydrogen as an energy substrate. H. hepaticus hydro...

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Immunogenic proteins ofHelicobacter pullorum,Helicobacter bilisandHelicobacter hepaticusidentified by two-dimensional gel electrophoresis and immunoblotting

Cited by 29 publications

References 24 publications

Cross-Reactivity between Immune Responses to Helicobacter bilis and Helicobacter pylori in a Population in Thailand at High Risk of Developing Cholangiocarcinoma

Cross-Reactivity between Immune Responses to Helicobacter bilis and Helicobacter pylori in a Population in Thailand at High Risk of Developing Cholangiocarcinoma

Increased prevalence of seropositivity for non-gastric Helicobacter species in patients with autoimmune liver disease

Helicobacter hepaticus Hydrogenase Mutants Are Deficient in Hydrogen-Supported Amino Acid Uptake and in Causing Liver Lesions in A/J Mice

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