The effect on enterotoxicity of protease purified from Vibrio cholerae O1

Ichinose, Y

doi:10.1016/0378-1097(94)90025-6

Cited by 10 publications

(11 citation statements)

References 14 publications

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“…5A). Our results are in agreement with the finding that pretreatment of ileal loops with purified V. cholerae HapA increased fluid accumulation in loops inoculated with live vibrios (20). The hapA and hapA motY mutants expressed less CT and TcpA (Fig.…”

Section: Discussionsupporting

confidence: 92%

Contribution of Hemagglutinin/Protease and Motility to the Pathogenesis of El Tor Biotype Cholera

et al. 2006

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Vibrio cholerae is a highly motile organism that secretes a Zn-dependent metalloprotease, hemagglutinin/protease (HapA). HapA has been shown to have mucinase activity and contribute to the reactogenicity of live vaccine candidates, but its role in cholera pathogenesis is not yet clear. The contribution of motility to pathogenesis is not fully understood, since conflicting results have been obtained with different strains, mutants, and animal models. The objective of this work was to determine the contribution of HapA and motility to the pathogenesis of El Tor biotype cholera. To this end we constructed isogenic motility (motY) and mucinase (hapA) single and double mutants of an El Tor biotype V. cholerae strain. Mutants were characterized for the expression of major virulence factors in vitro and in vivo. The motility mutant showed a remarkable increase in cholera toxin (CT), toxin coregulated pilus major subunit (TcpA), and HapA production in vitro. Increased TcpA and CT production could be explained by increased transcription of tcpA, ctxA, and toxT. No effect was detected on the transcription of hapA in the motility mutant. The sodium ionophore monensin diminished production of HapA in the parent but not in the motility mutant. Phenamil, a specific inhibitor of the flagellar motor, diminished CT production in the wild-type and motY strains. The hapA mutant showed increased binding to mucin. In contrast, the motY mutation diminished adherence to biotic and abiotic surfaces including mucin. Lack of HapA did not affect colonization in the suckling mouse model. The motility and mucinase defects did not prevent induction of ctxA and tcpA in the mouse intestine as measured by recombinasebased in vivo expression technology. Analysis of mutants in the rabbit ileal loop model showed that both V. cholerae motility and HapA were necessary for full expression of enterotoxicity.Cholera is an acute diarrheal disease characterized by the passing of voluminous rice water stool. Vibrio cholerae of serogroups O1 and O139 continues to cause seasonal cholera outbreaks that affect highly populated regions in Asia, Africa, and Latin America. V. cholerae is a highly motile organism with a single sheathed polar flagellum. It colonizes the small intestine and expresses a variety of virulence determinants, such as the toxin-coregulated pilus (TCP), cholera toxin (CT), and other factors required to multiply and survive in the host.V. cholerae produces a soluble Zn-metalloprotease, hemagglutinin/protease (HapA), encoded by hapA (18). HapA can proteolytically degrade several physiologically important host proteins including mucin (10). HapA perturbs the paracellular barrier of cultured intestinal epithelial cells (33, 46) by acting on tight junction-associated proteins (47). Inactivation of hapA increased adherence to mucin synthesized by HT29-18N2 cells (4), and expression of hapA was required for V. cholerae to penetrate a mucin-containing gel in vitro (42). Although analysis of hapA mutants in infant rabbits and suckling mice has not ...

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Section: Discussionsupporting

confidence: 92%

Contribution of Hemagglutinin/Protease and Motility to the Pathogenesis of El Tor Biotype Cholera

et al. 2006

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“…It also nicks and activates CT and CT-related enterotoxins [24]. Besides the protease activity, HA/P also shows hemagglutinating activity [25,26] on erythrocytes of Balb/c mice, but this activity has not been consistently reproduced [27,28]. Our study showed that NMDCY has a protease activity which is inhibited by metal chelators, but has no hemagglutinating activity on Balb/c mice and chicken erythrocytes.…”

Section: Resultsmentioning

confidence: 97%

Characterization of non-membrane-damaging cytotoxin of non-toxigenic Vibrio cholerae O1 and its relevance to disease

Mitra

1998

FEMS Microbiology Letters

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The non-membrane-damaging cytotoxin which causes dramatic cell rounding of cultured HeLa cells was purified to homogeneity from a clinical strain (WO5) of non-toxigenic Vibrio cholerae O1 Inaba belonging to the El Tor biotype. The purified protein has a denatured molecular weight of 35 kDa and a native molecular weight of approximately 37 kDa indicating the monomeric nature of the protein. The 15 N-terminal amino acid sequence of non-membrane-damaging cytotoxin showed complete homology to the hemagglutinin protease previously purified and characterized from V. cholerae O1. Purified nonmembrane-damaging cytotoxin from V. cholerae O1 was immunologically and biochemically identical to that previously purified from V. cholerae O26. Non-membrane-damaging cytotoxin was found to be enterotoxic in rabbit ileal loop assay inducing accumulation of non-hemorrhagic fluid at 100 Wg and elicited a concentration dependent increase in short circuit current and tissue conductance of rabbit ileal mucosa mounted on Ussing chambers. A significant serum immunoglobulin G response against non-membrane-damaging cytotoxin was elicited by patients infected with V. cholerae O139 but not with V. cholerae O1. These properties make non-membrane-damaging cytotoxin a potential virulence factor of V. cholerae which should be taken into consideration while making live, attenuated recombinant vaccine strains against cholera. z 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

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“…The RIL test was carried out using Japanese white rabbits weighing 2.0-2.5 kg by the method of Ichinose et al (13). Briefly, rabbits starved for 48 hr were anesthetized with sodium pentobarbital (20 mg kg-1 of body weight) and laparotomy was performed.…”

Section: Methodsmentioning

confidence: 99%

Detection of Genes Encoding Cholera Toxin (CT), Zonula Occludens Toxin (ZOT), Accessory Cholera Enterotoxin (ACE) and Heat‐Stable Enterotoxin (ST) in Vibrio mimicus Clinical Strains

Shi

Miyoshi

Hiura

et al. 1998

Microbiology and Immunology

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A total of 51 clinical strains of Vibrio mimicus were searched for the presence of virulence-associated genes, like ctx, zot or ace genes which locate in "cholera virulence cassette," and the st gene by polymerase chain reaction. Moreover, the pathological potential of each clinical strain was also examined by rabbit ileal loop (RIL). Three strains showed to have the ctx gene, of which only one strain was zot gene-positive. Meanwhile, one other strain was zot but ctx-. All of these four strains were found to have the ace gene and to belong to serogroup O115. Nine strains showed to carry the st gene. However, none of these ST-genepositive strains was indicated to contain the genes located in the "cholera virulence cassette." It is of interest to note that all of the RIL-positive and/or virulence gene-positive strains were restricted to three serogroups, O20, O41 and O115. These results suggest a significant association between O antigens and enterotoxic activities in V mimicus clinical strains, and clearly demonstrate multifactorial virulence potentials of this human pathogen.

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The effect on enterotoxicity of protease purified from Vibrio cholerae O1

Cited by 10 publications

References 14 publications

Contribution of Hemagglutinin/Protease and Motility to the Pathogenesis of El Tor Biotype Cholera

Contribution of Hemagglutinin/Protease and Motility to the Pathogenesis of El Tor Biotype Cholera

Characterization of non-membrane-damaging cytotoxin of non-toxigenic Vibrio cholerae O1 and its relevance to disease

Detection of Genes Encoding Cholera Toxin (CT), Zonula Occludens Toxin (ZOT), Accessory Cholera Enterotoxin (ACE) and Heat‐Stable Enterotoxin (ST) in Vibrio mimicus Clinical Strains

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