Purification and partial characterization of heat-stable enterotoxin of enterotoxigenic Escherichia coli

Takeda, Yoshifumi; Takeda, Tae; Yano, Takuo; Yamamoto, Kazuhiko; Miwatani, Toshio

doi:10.1128/iai.25.3.978-985.1979

Cited by 125 publications

(43 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Although many large plasmids have been reported to be involved in the pathogenicity to animal salmonellosis, our conclusion in the present study is that virulence factors of Salmonella causing human acute enterocolitis may not be coded by genes on a plasmid. The reasons are: (1) only 58% of 1,041 Salmonella isolates from patients contained plasmids most of which were heterogeneous in size; (2) although the virulence of S. typhimurium to cause mouse typhoid is believed to be determined by the 100-kb plasmid, 70% of the isolates from enterocolitis patients did not contain the plasmid ; (3) plasmid-eliminated strains, as well as naturally plasmid-free isolates, of S. braenderup and S. typhimurium showed virulence as strong as the plasmid-bearing parents in various pathogenicity tests for enteropathogenicity, which have been recognized to represent the potentials of many bacteria to cause human gastroenteritis (3,11,18,22). We postulate, therefore, that the virulence plasmids are involved only in a systemic infection such as mouse typhoid or septicemia and not in an infection such as human enterocolitis, the basis of which is not clear.…”

Section: Discussionmentioning

confidence: 99%

The 106‐Kilobase Plasmid of Salmonella braenderup and the 100‐Kilobase Plasmid of Salmonella typhimurium Are Not Necessary for the Pathogenicity in Experimental Models

Horiuchi

Goto

Inagaki

et al. 1991

Microbiology and Immunology

View full text Add to dashboard Cite

Among 1,041 clinical isolates (77 serovars) of Salmonella which had been derived from cases with acute enterocolitis, 601 (58°)) contained one or more plasmids. Large serovar-specific plasmids were seen in 95 of 307 isolates (31%) of Salmonella typhimurium, in 34 of 34 isolates (100%) of Salmonella enteritidis and in 36 of 38 isolates (94.7%) of Salmonella braenderup: the sizes of which were 100, 60 and 106 kilobases (kb), respectively.In order to determine the role of these plasmids in pathogenicity for enterocolitis, the plasmids were eliminated from some strains of S. braenderup and S. typhimurium and the pathogenicity of the plasmid-less strains was compared with that of the parent strains by invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, lesion of mucosal tissue and the Sereny test. The virulence of all the plasmid-less strains was as strong as that of the plasmidbearing strains in these pathogenicity assay systems. We therefore concluded that the 106-kb plasmid of S. braenderup and the 100-kb plasmid of S. typhimurium are not necessary for their pathogenicity in the experimental models : invasiveness to HeLa cells, fluid accumulation in the rabbit ligated ileal loop, and Sereny test.

show abstract

Section: Discussionmentioning

confidence: 99%

The 106‐Kilobase Plasmid of Salmonella braenderup and the 100‐Kilobase Plasmid of Salmonella typhimurium Are Not Necessary for the Pathogenicity in Experimental Models

Horiuchi

Goto

Inagaki

et al. 1991

Microbiology and Immunology

View full text Add to dashboard Cite

show abstract

“…The suckling mouse assay was essentially carried out as described by Takeda et al [18]. Namely, breeding colonies of white mice were obtained from SLC, Shizuoka, Japan.…”

Section: Suckling Mouse Assaymentioning

confidence: 99%

Isolation and characterization of a cytotoxin produced byPlesiomonas shigelloidesP-1 strain

Okawa

Ohtomo

Tsugawa

et al. 2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

In order to clarify the enteropathogenicity of Plesiomonas shigelloides, we investigated a cytotoxin produced by the P-1 strain isolated from patients suffering from diarrhea. The cytotoxicity of the culture filtrate of the strain reached a maximum in culture at 37 degrees C after 12 h shaken in BHI medium. The cytotoxin in the cultures was purified by (NH4)2SO4 precipitation, and Sephacryl S-100, Mono Q HR, and Superdex 200 HR column chromatographies. An approximate 340-fold purification was achieved, with a recovery of about 1.4%, from the culture supernatant. The cytotoxin is heat-stable, and is a complex of three major proteins (LPS-binding proteins with molecular weights of 32, 40, and 48 kDa), with lipopolysaccharide (LPS) giving a total a molecular weight of more than 600 kDa. The ratio of protein to LPS in the cytotoxin was 6-5. The cytotoxic activity was reduced by about 80% by proteinase K treatment or when incubated with anti-cholera toxin antibody (Anti-CT). Western blotting of the cytotoxin with Anti-CT demonstrated the presence of two anti-cholera toxin-reactive protein (ACRP) bands with molecular weights of 40 kDa (a major single protein band) and 48 kDa. The N-terminal amino acid sequence (20 residues) of the 40 kDa protein was 75% identical to Pasteurella multocida cell membrane proteins. The cytotoxin gave a positive reaction in the suckling mouse assay whereas LPS alone hardly exhibited any cytotoxic or enterotoxigenic activity. In conclusion, P. shigelloides produces a cytotoxin that consists of a complex of protein and LPS with the former component exhibiting both cytotoxicity and enteropathogenicity. This cytotoxin has the potential to have an important role in the enteropathogenicity of P. shigelloides.

show abstract

“…Intestinal loop test in mice. The intestinal loop test in mice was carried out essentially as described previously (36). Male ddY mice weighing 20 to 25 g were used without starvation.…”

Section: Fraction Numbermentioning

confidence: 99%

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Yamamoto¹,

Takeda²,

Miwatani³

et al. 1983

Infect Immun

Self Cite

View full text Add to dashboard Cite

Cholera-like enterotoxin was isolated and purified from the culture supernatant of a non-O1 strain of Vibrio cholerae, E8498, isolated from the environment. Enterotoxin was purified by aluminum hydroxide absorption and elution and successive gel filtrations on Sephadex G-100, Bio-Gel A-5m, and Sephadex G-75. Purified enterotoxin gave a single stained band on polyacrylamide gel disc electrophoresis, and the mobility was the same as that of cholera enterotoxin. The specific biological activity of the purified enterotoxin was almost the same as that of cholera enterotoxin in the Chinese hamster ovary cell assay, fluid accumulation in mouse ligated intestine, increase in vascular permeability in rabbit skin, and passive immune hemolysis. Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis showed that the purified enterotoxin consisted of subunits A and B, identical to those of cholera enterotoxin, and Ouchterlony double gel diffusion tests indicated that the two toxins were immunologically identical. Enterotoxins prepared from several non-O1 strains isolated from human patients were also immunologically identical to cholera enterotoxin.Sodium azide and sodium EDTA were added to culture supernatants at final concentrations of 0.02 and 0.05%, respectively.Purified CT. CT from V. cholerae 569B, serovar 01, purified by the method described by Ohtomo et al. 1128 on July 15, 2020 by guest http://iai.asm.org/ Downloaded from

show abstract

Purification and partial characterization of heat-stable enterotoxin of enterotoxigenic Escherichia coli

Cited by 125 publications

References 22 publications

The 106‐Kilobase Plasmid of Salmonella braenderup and the 100‐Kilobase Plasmid of Salmonella typhimurium Are Not Necessary for the Pathogenicity in Experimental Models

The 106‐Kilobase Plasmid of Salmonella braenderup and the 100‐Kilobase Plasmid of Salmonella typhimurium Are Not Necessary for the Pathogenicity in Experimental Models

Isolation and characterization of a cytotoxin produced byPlesiomonas shigelloidesP-1 strain

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Contact Info

Product

Resources

About