Mechanisms of chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus in human colonic tissue and a human intestinal epithelial cell line

Takahashi, Aline Barral; Sato, Yasuomi D.; Shiomi, Yoshihiko; Cantarelli, Vlademir Vicente; Iida, Tetsuya; Lee, M.; Honda, Takeshi

doi:10.1099/0022-1317-49-9-801

Cited by 49 publications

(38 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Unlike that of TDH, the activity of TRH is labile to heat treatment at 60°C for 10 min (10). Although we recently demonstrated that TDH induced Cl Ϫ secretion in human colonic epithelial cells (22), there is no evidence how TRH induces fluid secretion. The present study was designed to investigate how TRH affects short-circuit current (Isc) in human colonic epithelial cell (Caco-2 cell) monolayers and the intracellular Ca 2ϩ concentration ([Ca 2ϩ ] in ) in Caco-2 cells after exposure to TRH.…”

mentioning

confidence: 92%

“…The present study was designed to investigate how TRH affects short-circuit current (Isc) in human colonic epithelial cell (Caco-2 cell) monolayers and the intracellular Ca 2ϩ concentration ([Ca 2ϩ ] in ) in Caco-2 cells after exposure to TRH. The measurements of Isc and [Ca 2ϩ ] in in Caco-2 cells were taken by following a previously described method (22). Briefly, stock cells were trypsinized, suspended at 20 ϫ 10 4 cells/ml in medium, and seeded at confluent density onto 1.0-cm 2 transwell inserts (Costar, Cambridge, Mass.).…”

mentioning

confidence: 99%

“…[Ca 2ϩ ] in was determined by microfluo- rometry using a fluorescent dye, 1-(2-(5Ј-carboxyoxazol-2Ј-yl)-6-aminobenzofuran-5-oxy)-2(2Јamino-5Ј-methylphenoxy) ethane-N,N,NЈ,NЈ-tetraacetic acid, pentaacetoxy methyl ester (Fura-2/AM; Molecular Probes, Eugene, Ore.). Using a previously described method (22,23), TRH was purified from a clinically isolated strain of V. parahaemolyticus (RIMD 2210531), which was KP negative (TDH negative) and yielded a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Fig. 1A), suggesting that the toxins were purified to homogeneity.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

et al. 2000

View full text Add to dashboard Cite

A hemolytic toxin related to thermostable direct hemolysin (TDH), TDH-related hemolysin (TRH), produced by Kanagawa-phenomenon-negative Vibrio parahaemolyticus is suspected of playing an important, but yet-tobe-elucidated role in diarrhea caused by this organism. In cultured human colonic epithelial cells, TRH increases Cl ؊ secretion, followed by elevation of intracellular calcium.Vibrio parahaemolyticus is an important cause of gastroenteritis, with outbreaks now occurring all over the world (2, 17). However, the mechanisms underlying the diarrheic action of this pathogen are not completely understood. V. parahaemolyticus can be categorized according to the presence or absence of the Kanagawa phenomenon (KP), in which there is hemolysis due to thermostable direct hemolysin (TDH) produced by the organism on Wagatsuma agar medium (14). TDH has been considered a major virulence factor for this disease (8, 23); however, some cases of V. parahaemolyticus gastroenteritis are due to KP-negative strains (10,15,16). From clinically isolated KP-negative V. parahaemolyticus, we identified a new hemolysin, named TDH-related hemolysin (TRH), and demonstrated that this TRH was also an important virulence factor (10, 24). For example, TRH stimulated fluid secretion in the rabbit loop test (8, 10). Thus, there is a possibility that TRH induces diarrhea. TRH is immunologically related, but not identical, to TDH ordinary human virulence factor. Unlike that of TDH, the activity of TRH is labile to heat treatment at 60°C for 10 min (10). Although we recently demonstrated that TDH induced Cl Ϫ secretion in human colonic epithelial cells (22), there is no evidence how TRH induces fluid secretion. (175, 83, 62, 47.5, 32.5, 25, 16.5, and 6.5 kDa); lane 2, TRH. (B) Effect of gluconate ion and Ca 2ϩ depletion on Isc in Caco-2 cell monolayers. F, Control cells (simple exposure to TRH); s, Cl Ϫ replaced by gluconate ion in bath solution of Ussing chamber; OE, CaCl 2 was omitted from bath solution and EGTA (1 mM) was added to bath solution of Ussing chamber. TRH was added on the apical side of the Caco-2 cells at the time indicated. , negative control (no exposure to TRH). Values are expressed as means Ϯ standard deviations (n ϭ 5). ‫,ء‬ Significant difference (P Ͻ 0.05) versus negative control.

show abstract

mentioning

confidence: 92%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

et al. 2000

View full text Add to dashboard Cite

show abstract

“…Thus, TDH effects on epithelial cells are important for biological functions, such as diarrhea. The addition of TDH to the mucosal side of human colonic tissue in Ussing chambers led to increased short circuit currents (Isc), a process that was inhibited by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), an inhibitor of Ca2+-activated chloride (Cl (20).…”

Section: Thermostable Direct Hemolysin (Tdh) and Tdh-related Hemolysimentioning

confidence: 99%

“…! for 10 min (20). Both Vp-TDH and Vp-TRH induce chloride secretion in human colonic epithelial cells (20,23).…”

Section: Thermostable Direct Hemolysin (Tdh) and Tdh-related Hemolysimentioning

confidence: 99%

Diarrhea induced by infection of <I>Vibrio parahaemolyticus</I>

Shimohata

Takahashi

2010

J. Med. Invest.

View full text Add to dashboard Cite

Many species of bacteria induce secretory and inflammatory diarrhea (1). The increased intestinal fluid secretion in diarrhea appears to result from the active secretion of chloride (1-3), a principal anion, but the way in which diarrhea is caused by individual bacterial infections has not been completely elucidated. Vibrio parahaemolyticus is a Gram-negative halophilic bacterium that naturally occurs in marine and estuarine environments (4). It is a human pathogen that causes food-borne acute gastroenteritis, often associated with the consumption of raw or undercooked seafood (5, 6). Clinical symptoms of V. parahaemolyticus infections include watery diarrhea, abdominal cramps, nausea, vomiting, headaches, fever, and chills (7,8). THERMOSTABLE DIRECT HEMOLYSIN (TDH) AND TDH-RELATED HEMOLYSIN (TRH)The majority of V. parahaemolyticus clinical isolates from patients with diarrhea has produced TDH and/or TRH, which are encoded by the tdh and trh genes, respectively (9). Strong associations have been found between gastroenteritis and these two proteins (10, 11). Therefore, TDH and TRH are regarded as major virulence factors of V. parahaemolyticus.V. parahaemolyticus TDH (Vp-TDH) is a proteinaceous toxin composed of 165 amino acids with one disulfide bond near the carboxyl terminus (12). The protein is a dimer, which lacks lipid and carbohydrate moieties, and has a molecular weight of c. 42 kDa by gel filtration and 21 kDa by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (13,14). Vp-TDH exhibits β-hemolytic activity on Wagatsuma medium, which has been termed the Kanagawa phenomenon (KP). Purified Vp-TDH is heat-stable, even at 100! !for REVIEW Diarrhea induced by infection of Vibrio parahaemolyticusTakaaki Shimohata and Akira Takahashi Department of Preventive Environment and Nutrition, Institute of Health Biosciences, the University of Tokushima Graduate School, Tokushima, JapanAbstract : Vibrio parahaemolyticus is a human pathogen that naturally inhabits marine and estuarine environments. Infection with V. parahaemolyticus is often associated with the consumption of raw or undercooked seafood, causing gastroenteritis with watery diarrhea. The presence of two type III secretion system (T3SS) proteins, thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH), has been closely associated with the severity of diarrheal illness. TDH and TRH have various biological activities including hemolytic activity, cardiotoxicity, and enterotoxicity. T3SS1 is involved in cytotoxicity to host cells and orchestrates a multifaceted host cell infection by induction of autophagy, cell rounding, and cell lysis. T3SS2 is thought to be related to the enterotoxicity of V. parahaemolyticus. The activities of inducing diarrhea of each of the virulence factors were summarized in this review.

show abstract

Vibrio parahaemolyticus—Seafood Safety and Associations with Higher Organisms

Qadri

Chowdhury

Takeda

et al.

Oceans and Health: Pathogens in the Marine Environment

View full text Add to dashboard Cite

Mechanisms of chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus in human colonic tissue and a human intestinal epithelial cell line

Cited by 49 publications

References 24 publications

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

Diarrhea induced by infection of <I>Vibrio parahaemolyticus</I>

Vibrio parahaemolyticus—Seafood Safety and Associations with Higher Organisms

Contact Info

Product

Resources

About

Mechanisms of chloride secretion induced by thermostable direct haemolysin of Vibrio parahaemolyticus in human colonic tissue and a human intestinal epithelial cell line

Cited by 49 publications

References 24 publications

Cl − Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

Cl − Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

Diarrhea induced by infection of <I>Vibrio parahaemolyticus</I>

Vibrio parahaemolyticus—Seafood Safety and Associations with Higher Organisms

Contact Info

Product

Resources

About

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin

Cl ⁻ Secretion in Colonic Epithelial Cells Induced by the Vibrio parahaemolyticus Hemolytic Toxin Related to Thermostable Direct Hemolysin