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1.
Inability of an isogenic urease-negative mutant stain of Helicobacter mustelae to colonize the ferret stomach. 总被引:1,自引:6,他引:1
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K A Andrutis J G Fox D B Schauer R P Marini J C Murphy L Yan J V Solnick 《Infection and immunity》1995,63(9):3722-3725
Eight ferrets specific-pathogen-free for Helicobacter mustelae were given, per dose, approximately 3.0 x 10(7) CFU of either the wild-type parent strain of H. mustelae (NCTC 12032) (two ferrets) the isogenic urease-negative mutant strain of H. mustelae (10::Tn3Km) (four ferrets), or sterile culture broth (two ferrets). Infection status was monitored by endoscopic gastric biopsy for urease activity, histopathology, and culture and by serology at 3, 6, 10, and 21 weeks. All ferrets were necropsied at 25 weeks. Both negative control ferrets remained uninfected, both ferrets receiving the H. mustelae wild-type parent strain became infected after two doses of the organism, and all four ferrets given two doses of the isogenic urease-negative mutant strain of H. mustelae remained uninfected throughout the 6-month study. Histopathology correlated with infection status. H. mustelae-infected ferrets exhibited diffuse mononuclear inflammation in the subglandular region and the lamina propria of the gastric mucosa, while uninfected ferrets showed no or minimal inflammation. These results suggest that urease activity is essential for colonization of the ferret stomach by H. mustelae. 相似文献
2.
Failure of surface ring mutant strains of Helicobacter mustelae to persistently infect the ferret stomach
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Patterson MM O'Toole PW Forester NT Noonan B Trust TJ Xu S Taylor NS Marini RP Ihrig MM Fox JG 《Infection and immunity》2003,71(5):2350-2355
Helicobacter mustelae, the gastric pathogen of ferrets, produces an array of surface ring structures which have not been described for any other member of the genus Helicobacter, including H. pylori. The unique ring structures are composed of a protein named Hsr. To investigate whether the Hsr rings are important for colonization of the ferret stomach, ferrets specific pathogen free for H. mustelae were inoculated with an Hsr-deficient mutant strain or the wild-type H. mustelae strain. Quantitative cultures from antral biopsy specimens obtained at 3, 6, and 9 weeks postinoculation demonstrated no significant difference in the levels of bacteria in the ferrets that received the Hsr-negative strain and the ferrets infected with the parent strain. However, when the ferrets were biopsied at 12 and 15 weeks and necropsied at 18 weeks after infection, the levels of bacteria of the Hsr-negative strain in the stomach antrum were significantly reduced. This decline contrasted the robust antral colonization by the wild-type strain. The Hsr-negative strain did not efficiently colonize the gastric body of the study ferrets. Histological examination at 18 weeks postinoculation revealed minimal gastric inflammation in the animals that received the mutant H. mustelae strain, a finding consistent with its waning infection status, whereas lesions characteristic of helicobacter infection were present in ferrets infected with the wild-type strain. Scant colonization by the Hsr-negative H. mustelae strain at the end of the 18-week study, despite initial successful colonization, indicates an inability of the mutant to persist, perhaps due to a specific host response. 相似文献
3.
Construction and characterization of an isogenic urease-negative mutant of Helicobacter mustelae.
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J V Solnick C Josenhans S Suerbaum L S Tompkins A Labigne 《Infection and immunity》1995,63(9):3718-3721
Helicobacter mustelae infects the ferret stomach and provides an opportunity to study pathogenic determinants of a Helicobacter species in its natural host. We constructed an isogenic urease-negative mutant of H. mustelae which produced no detectable urease and showed a reduced acid tolerance. This mutant provides an opportunity to further evaluate the role of urease in the pathogenesis of Helicobacter infection. 相似文献
4.
Adherence of isogenic flagellum-negative mutants of Helicobacter pylori and Helicobacter mustelae to human and ferret gastric epithelial cells
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Isogenic flagellum-negative mutants of Helicobacter pylori and Helicobacter mustelae were screened for their ability to adhere to primary human and ferret gastric epithelial cells, respectively. We also evaluated the adherence of an H. pylori strain with a mutation in the flbA gene, a homologue of the flbF/lcrD family of genes known to be involved in the regulation of H. pylori flagellar biosynthesis. H. pylori and H. mustelae mutants deficient in production of FlaA or FlaB and mutants deficient in the production of both FlaA and FlaB showed no reduction in adherence to primary human or ferret gastric epithelial cells compared with the wild-type parental strains. However, adherence of the H. pylori flbA mutant to human gastric cells was significantly reduced compared to the adherence of the wild-type strain. These results show that flagella do not play a direct role in promoting adherence of H. pylori or H. mustelae to gastric epithelial cells. However, genes involved in the regulation of H. pylori flagellar biosynthesis may also regulate the production of an adhesin. 相似文献
5.
On the basis of analysis of protein profiles, isolates of Helicobacter pylori and Helicobacter mustelae were less than 40% similar. Cytotoxin produced by H. pylori was not detected in isolates of H. mustelae. Both bacterial species agglutinated human erythrocytes. These results substantiate a taxonomic difference between H. pylori and H. mustelae. 相似文献
6.
Helicobacter mustelae is a urease-rich bacterium associated with gastritis in ferrets. The ureases of H. mustelae and Helicobacter pylori, a bacterium implicated in human gastritis, share many characteristics. Helicobacter sp. ureases appear to be unique among bacterial enzymes in exhibiting submillimolar Km values and in being composed of two subunits. 相似文献
7.
Induction of interleukin-8 secretion from gastric epithelial cells by a cagA negative isogenic mutant of Helicobacter pylori. 总被引:14,自引:3,他引:14
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J E Crabtree Z Xiang I J Lindley D S Tompkins R Rappuoli A Covacci 《Journal of clinical pathology》1995,48(10):967-969
The ability of Helicobacter pylori strains to induce interleukin-8 (IL-8) gene expression and protein secretion from gastric epithelial cell lines in vitro is variable. This cellular response is associated with bacterial expression of the CagA protein present in type I H pylori strains. To determine the role of CagA in this host cell response, an isogenic cagA negative mutant, N6.XA3, was constructed. The cagA negative isogenic mutant and the wild-type parental cagA positive strain, N6, were cocultured with AGS, ST-42 and KATO-3 gastric epithelial cell lines and secreted interleukin-8 assayed by enzyme linked immunosorbent assay. In all three cell lines there was no significant difference in the IL-8 secretion induced by the cagA negative isogenic mutant, N6.XA3, and the wild-type parent strain, N6. These studies show that CagA is not the inducer of IL-8 secretion from gastric epithelial cells. As all wild-type CagA positive strains studied to date induce IL-8, the bacterial factor(s) inducing this inflammatory response is closely associated with the expression of CagA. 相似文献
8.
Helicobacter mustelae and Helicobacter pylori bind to common lipid receptors in vitro. 总被引:9,自引:6,他引:3
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Helicobacter pylori is a recently recognized human pathogen causing chronic-active gastritis in association with duodenal ulcers and gastric cancer. Helicobacter mustelae is a closely related bacterium with similar biochemical and morphologic characteristics. H. mustelae infection of antral and fundic mucosa in adult ferrets causes chronic gastritis. An essential virulence property of both Helicobacter species is bacterial adhesion to mucosal surfaces. The aim of this study was to determine whether H. mustelae binds to the same lipids shown previously to be receptors for H. pylori adhesion in vitro. By using thin-layer chromatography overlay and a receptor-based enzyme-linked immunosorbent assay, H. mustelae was found to bind the same receptor lipids as H. pylori, namely, phosphatidylethanolamine and gangliotetraosylceramide. In addition, both H. pylori and H. mustelae bound to a deacylplasmalogen phosphatidylethanolamine. In contrast to H. pylori, H. mustelae binding to receptors was unaffected by motility or viability. Murine monoclonal and bovine polyclonal antibodies against exoenzyme S, and exoenzyme S itself (from Pseudomonas aeruginosa), inhibited binding of H. mustelae to phosphatidylethanolamine and gangliotetraosylceramide. These findings show that H. mustelae binds in vitro to the same lipid receptors as H. pylori and suggest that the adhesion of H. mustelae to such species is mediated by preformed, surface-exposed adhesins which include an exoenzyme S-like protein. 相似文献
9.
The fatty acid compositions of Helicobacter mustelae whole cells, isolated phospholipids, and isolated lipopolysaccharides were analyzed by gas-liquid chromatography. Major phospholipid fatty acids were C16:0, C18:0, C18:1, and C19:0 cyc. In isolated lipopolysaccharides, 3-OH-C16:0, 3-OH-C14:0, C14:0, C16:0, and C18:0 were found. The lipid composition of H. mustelae thus showed pronounced differences from that of H. pylori. Flagella were purified by mechanical shearing and centrifugation steps. In all H. mustelae strains, the flagellin had an apparent molecular mass of 53 kDa and was thus the same size as H. pylori flagellin. The flagellin of strain NCTC 12032 was further purified and subjected to N-terminal amino acid sequence analysis. The first 10 amino acids were identical to those of H. pylori flagellin, but the next 5 were different. Significant homology was also found with flagellins of other bacteria. 相似文献
10.
González-Valencia G Muñoz-Perez L Morales-Espinosa R Camorlinga-Ponce M Muñoz O Torres J 《Journal of clinical microbiology》2008,46(8):2783-2785
The diversity in the expression of Lewis antigens (Le) of 226 single colonies of Helicobacter pylori isolated from four regions of the stomach of eight adults is shown. Ley was expressed more in strains colonizing antrum than in strains colonizing fundus, whereas Lex was more common in fundus strains. cagA+ strains were more associated with Le-negative strains. 相似文献
11.
A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. 总被引:10,自引:7,他引:10
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The urease of Helicobacter pylori has been proposed to be one of its pathogenic factors. A kanamycin resistance determinant was inserted in a cloned urease gene, and transformation-mediated allelic exchange mutagenesis was carried out to introduce the disrupted gene into the corresponding wild-type chromosomal region of a clinical isolate of H. pylori, CPY3401. The resulting mutant, HPT73, had the null activity of urease. Nude mouse stomachs were challenged with these two isogenic strains to examine the role of urease in pathogenesis. Gastritis was found in the CPY3401-challenged stomachs, from which bacteria indistinguishable from CPY3401 were recovered. There was no gastritis in the HPT73-challenged stomachs, and we could not recover H. pylori from them. These results indicated that H. pylori urease is essential for colonizing the nude mouse stomach. 相似文献
12.
Helicobacter mustelae isolation from feces of ferrets: evidence to support fecal-oral transmission of a gastric Helicobacter. 总被引:9,自引:0,他引:9
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J G Fox B J Paster F E Dewhirst N S Taylor L L Yan P J Macuch L M Chmura 《Infection and immunity》1992,60(2):606-611
Helicobacter mustelae has been isolated from stomachs of ferrets with chronic gastritis and ulcers. When H. mustelae is inoculated orally into H. mustelae-negative ferrets, the animals become colonized and develop gastritis, a significant immune response, and a transient hypochlorhydria. All of these features mimic Helicobacter pylori-induced gastric disease in humans. Because the epidemiology of H. pylori infection is poorly understood and its route of transmission is unknown, the feces of weanling and adult ferrets were cultured for the presence of H. mustelae. H. mustelae was isolated from the feces of 11 of 36 ferrets by using standard helicobacter isolation techniques. H. mustelae was identified by biochemical tests, ultrastructural morphology, reactivity with specific DNA probes, and 16S rRNA sequencing. H. mustelae was not recovered from 20-week-old ferrets which had been H. mustelae positive as weanlings, nor was H. mustelae recovered from 1-year-old ferrets. Isolation of H. mustelae from feces may correspond to periods of transient hypochlorhydria, or H. mustelae may be shed in feces intermittently. The H. mustelae-colonized ferret provides an ideal model for studying the pathogenesis and transmission of H. pylori-induced gastric disease. 相似文献
13.
Protein Hpn: cloning and characterization of a histidine-rich metal-binding polypeptide in Helicobacter pylori and Helicobacter mustelae. 总被引:3,自引:1,他引:2
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J V Gilbert J Ramakrishna F W Sunderman Jr A Wright A G Plaut 《Infection and immunity》1995,63(7):2682-2688
Helicobacter pylori is a human gastrointestinal pathogen involved in gastritis, duodenal ulcers, and gastric neoplasia. This microorganism produces large amounts of a urease which, like all known ureases, has nickel in the active site. We have identified a protein in clinical isolates of H. pylori and an identical protein in the ferret pathogen Helicobacter mustelae that strongly binds Ni2+ and Zn2+. This protein has been named Hpn to emphasize its origins in H. pylori and its affinity for nickel. The encoding hpn gene, cloned and expressed in Escherichia coli ER1793, has an open reading frame (180 bp) that specifies a protein with a calculated molecular mass of 7,077 Da and with the same amino-terminal sequence as that of wild-type Hpn. The deduced sequence of Hpn consists of 60 amino acids, of which 28 (47%) are histidines. The hpn gene does not map with the urease gene cluster on the H. pylori chromosome. An Hpn-negative, isogenic H. pylori strain, generated by hpn gene deletion and grown on blood agar, had the same urease activity that wild-type cells did. Thus, the role of Hpn in helicobacters is unknown. 相似文献
14.
Sequence and antigenic variability of the Helicobacter mustelae surface ring protein Hsr 总被引:1,自引:0,他引:1
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We have identified an array of more than 500 repetitive sequences flanking the hsr gene, which encodes the major surface protein of the ferret pathogen Helicobacter mustelae. The repeats show identity exclusively to the amino-terminal half of Hsr. Analysis of Hsr from three strains indicated variability of exposed epitopes. Characterization of an hsr mutant showed that Hsr is not an adhesin. 相似文献
15.
Heiji Kumazawa Hiroshi Takagi Katsuko Sudo Wataru Nakamura Syun Hosoda 《Virchows Archiv : an international journal of pathology》1989,416(2):141-151
Summary Neoplastic and nonneoplastic lesions developing spontaneously in antral and fundic mucosae of stomachs of mutant chamois-coloured Z (130 animals) and Y (67 animals) strains of Mastomys aged 18 to 24 months were examined histologically and histochemically. The Z strain developed both antral lesions (hyperplasia 29.2%; dysplasia 23.8%; adenocarcinoma 17.7%) and fundic carcinoid(s) (72.3%). The antral lesions were limited to the lesser curvature near the pyloric ring. Macroscopically, adenocarcinomas resembled human gastric carcinomas of either Borrmann's type I or II. Histochemically, adenocarcinoma cells were characterised by marked reduction of total mucins produced and predominance of mucins with both periodic acid-Schiff and Alcian blue reactivities (neutral and sialated class II mucins). An infiltrating adenocarcinoma was successfully transplanted into nude mice, reaching the 7th generation of transplantations over 4 years, and retained histological features of the primary tumour. The ultrastructural appearance of growing transplanted tumours supported the reduced production of mucins by adenocarcinoma cells with scarcity of mucin granules and intracellular cysts.However, the Y strain never developed antral lesions like the wild strain, developing fundic carcinoid(s) only. Microscopically, these carcinoids contained argyrophilic nonargentaffin granules, and biochemically produced histamine consistently but no 5-hydroxytryptamine (5-HT, serotonin) like those of the wild strain. Since we found unexpectedly that a line of F2 but not F1 hybrids between wild and Z strains developed the same antral lesions as Z strain, a preliminary experiment was performed to confirm the development of antral lesions in F2 hybrids newly produced by brother-sister mating. Among 41 surviving F2 offspring, 4 (9.8%) developed hyperplasia, 2 (4.9%) dysplasia and none adenocarcinoma. The numbers (6) of animals observed with these lesions approximated to their expected numbers (7.3). 相似文献
16.
The ultrastructure of Helicobacter mustelae, a natural inhabitant of the ferret stomach, has been studied and compared with the human gastroduodenal pathogen H. pylori. H. mustelae is a short, slightly curved rod, 2 microns x 0.5 micron, with four or more sheathed flagella. The most common flagellar configuration is a single flagellum at one terminus, bipolar arrangement at the other end and a lateral flagellum. Dense inclusion bodies were observed below the flagellar insertion sites. It is suggested that this configuration is a specialised adaptation to motility in a viscous environment. On examination of the ferret gastric mucosa, similarities to H. pylori were observed such as adherence to gastric tissue and the formation of adhesion pedestals. However, unlike H. pylori, significant numbers of bacteria were intracellular. Furthermore, a much greater proportion of H. mustelae were attached to the mucosa with few bacteria lying free in the mucus, as is seen with H. pylori. It is concluded that the ferret is an important model for the study of adherence of bacteria to gastric mucosa and their possible role in peptic ulceration. 相似文献
17.
18.
Antagonisms among isogenic strains of Escherichia coli in the digestive tracts of gnotobiotic mice. 总被引:9,自引:4,他引:5
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We have observed that antagonisms occur between isogenic strains of Escherichia coli associated with gnotobiotic mice. The strains differed in the carriage of plasmids or in chromosomal mutations. The plasmid-free strains, in general, inhibited the establishment of plasmid-bearing strains in the gastrointestinal tract of mice. The outcome of the interactions between isogenic pairs, however, depended on the order in which the strains were introduced into the mice. Maintaining the bacterial strains in monoassociation with gnotobiotic mice resulted in the "adaptation" of the bacteria to their host. Thus, in all cases, "adapted" strains became the dominant population in the feces of mice, regardless of whether the adapted strains was introduced into mice before or after its isogenic partner which had been cultured in vitro. The ecological advantage disappeared when the adapted strain was cultured in broth. Ultrastructural differences in cell morphology were observed between strains maintained in vivo and in vitro. 相似文献
19.
Campylobacter pylori subsp. mustelae was cultured from both normal and inflamed gastric mucosa of ferrets. Examination of neonatal, juvenile, and adult ferrets established that the gastric mucosa in the majority of preweanling (age, less than 6 weeks) ferrets sampled were not colonized with C. pylori subsp. mustelae, whereas the gastric mucosa of 100% of adult ferrets were colonized with this gastric organism. C. pylori subsp. mustelae was isolated from the gastric mucosa on a sequential basis from nine ferrets during a several-month period, inferring either persistent colonization or frequent reinfection with C. pylori subsp. mustelae. 相似文献
20.
Infection of adult Syrian hamsters with flagellar variants of Campylobacter jejuni. 总被引:7,自引:5,他引:7
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Two variants of Campylobacter jejuni IN1 differing in the expression of flagella, IN1 (Fla+ Mot+, wild type) and IN1-NM (Fla- Mot-), were tested for their ability to establish infection in adult hamsters. Animals were challenged intracecally with 2 X 10(9) to 5 X 10(10) CFU and monitored for evidence of infection. None of the challenged animals developed illness. There was a significant difference, however, in the ability of IN1 to infect hamsters (35 of 43) compared with that of IN1-NM (1 of 42) (P less than 0.01). Additionally, eight animals challenged with IN1-NM excreted only the campylobacters of Fla+ phenotype, which were shown to be identical with the parental Fla+ Mot+ type. Both groups of animals developed serum immunoglobulin G antibodies to outer membrane proteins of IN1 as measured by an enzyme-linked immunosorbent assay; however, only animals that excreted Fla+ organisms developed antiflagellin antibodies. In vitro reversibility from Fla+ to Fla- occurred with a frequency of 9.2 X 10(-6) per cell per generation; however, reversion from Fla- to Fla+ could not be detected in vitro. Further characterization of IN1-NM showed that it produced a cytoplasmic 62K flagellin subunit protein, but this protein lacked six epitopes detected in IN1 and also differed in its two-dimensional gel electrophoresis pattern. The results of these studies support the concept that an intact flagellum is necessary for intestinal infection with C. jejuni and that this model may be useful for studying the immune response to C. jejuni. 相似文献