Purification of exfoliative toxin produced by Staphylococcus hyicus and its antigenicity

Tanabe, Taishi; Sato, Hisaaki; Kuramoto, Masahiko; Saito, Hiroshi

doi:10.1128/iai.61.7.2973-2977.1993

Cited by 30 publications

(46 citation statements)

References 13 publications

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“…Studies have shown that both virulent and avirulent S. hyicus can be isolated from diseased pigs [1]. The virulent strains produce an approximately 27-kDa [2,3] or 30-kDa [1,4] exfoliative toxin. S. hyicus ex-foliative toxin has been shown to be the factor responsible for the alterations of the skin in EE in pigs [4] and the production of exfoliative toxin correlates with the ability of the strains to induce EE in pigs [5].…”

Section: Introductionmentioning

confidence: 99%

“…Patients with SSSS are usually under the age of 5 years [6] and EE a¡ects piglets in the age of 1^5 weeks, but mild forms of the disease can be observed among older pigs [7]. The histopathological alterations of the skin in the two diseases are similar [6,8] and are in both cases caused by toxins [3,4,9]. SSSS in humans is caused by infection with Staphylococcus aureus producing antigenically distinct exfoliative toxins ETA and ETB, which are the factors that cause exfoliation of the skin [9].…”

Section: Introductionmentioning

confidence: 99%

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Differentiation and distribution of three types of exfoliative toxin produced by Staphylococcus hyicus from pigs with exudative epidermitis

Andresen¹

1998

FEMS Immunology and Medical Microbiology

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Exfoliative toxins of approximately 30 kDa produced by Staphylococcus hyicus strains NCTC 10350, 1289D-88 and 842A-88 were purified and specific polyclonal antisera were raised against each of the toxins. It was shown by immunoblot analysis and ELISA that three exfoliative toxins from S. hyicus were antigenically distinct. The three toxins were designated ExhA, ExhB and ExhC. From 60 diseased pigs, each representing an outbreak of exudative epidermitis, a total of 584 isolates of S. hyicus were phage typed and tested for production of exfoliative toxin. ExhA-, ExhB- and ExhC-producing S. hyicus isolates were found in 12 (20%), 20 (33%) and 11 (18%), respectively, of the 60 pig herds investigated. Production of the different types of exfoliative toxin was predominantly associated with certain phage groups. However, toxin production was found in all of the six phage groups defined by the phage typing system. Some changes in the distribution of isolates between phage groups were observed when the results of this study were compared to previous investigations. In this study two new antigenically distinct exfoliative toxins were isolated and tools for in vitro detection of toxin producing S. hyicus isolates and for further studies on the exfoliative toxins from S. hyicus have been provided.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differentiation and distribution of three types of exfoliative toxin produced by Staphylococcus hyicus from pigs with exudative epidermitis

Andresen¹

1998

FEMS Immunology and Medical Microbiology

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“…Exudative epidermitis (EE) in pigs is caused by infection with certain strains of Staphylococcus hyicus [1]. Previous studies have shown that the factor responsible for the alterations of the skin in EE is a protein toxin of approximately 27 kDa [2] or 30 kDa [1,3], designated exfoliative toxin. This toxin is produced by virulent strains of S. hyicus [4,5].…”

Section: Introductionmentioning

confidence: 99%

Studies on the effect of divalent metal ions on exfoliative toxins from Staphylococcus hyicus: indications of ExhA and ExhB being metalloproteins

Andresen¹

1999

FEMS Immunology and Medical Microbiology

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The exfoliative toxins ExhA and ExhB produced by Staphylococcus hyicus strains NCTC10350 and 1289D-88, respectively, were investigated with regard to the effect of divalent metal ions on toxin production as measured in indirect enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies. Data were obtained as endpoint titer values and used as semiquantitative measures for the amount of exfoliative toxin detected in culture supernatants. It was shown that the endpoint titers of ExhA in supernatants from cultures of strain NCTC10350 grown in the presence of 0.5 mM CaCl2, Cu(NO3)2 or ZnSO4 were higher compared to titers obtained by growth in medium supplemented with a number of other divalent metal salts. The titer of ExhB as determined in the indirect ELISA was increased by addition of 0.5 mM CoCl2, Cu(NO3)2 or CuSO4 to the growth medium. When ExhA or ExhB, prepared without addition of metal salt to the liquid growth medium, was subsequently incubated with 25 mM of Co2+, Cu2+ or Zn2+, the endpoint titers of the toxins were increased. Dialysis of ExhA and ExhB prepared with Zn2+ and Co2+, respectively, against certain metal chelators, resulted in a reduction of the titer determined in ELISA. Other metal chelators had varied effect in the detection of the toxins in ELISA. It was, however, not possible to restore the recognition of toxins by the monoclonal antibodies by incubation of EDDHA-dialyzed toxin preparations with Co2+, Cu2+ or Zn2+. The results of this study suggest that ExhA and ExhB are metalloproteins.

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“…In the same year, we also reported that piglets and young chickens are susceptible to shET and that shET causes a rounding effect without cell death in the cultured cells (11). Recently, we purified shET from the culture filtrate of S. hyicus strain P-1 by ammonium sulfate precipitation, DEAE-cellulofine column chromatography, Sephadex G-75 gel filtration and polyacrylamide gel electrophoresis (16). We also reported some characteristics of purified shET (16).…”

mentioning

confidence: 95%

“…Recently, we purified shET from the culture filtrate of S. hyicus strain P-1 by ammonium sulfate precipitation, DEAE-cellulofine column chromatography, Sephadex G-75 gel filtration and polyacrylamide gel electrophoresis (16). We also reported some characteristics of purified shET (16). The molecular mass of shET estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is 27,000.…”

mentioning

confidence: 99%

Optimum Culture Conditions for Production of Exfoliative Toxin by Staphylococcus hyicus

Watanabe

Tanabe

Sato

et al. 1995

Microbiology and Immunology

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Optimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined. High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S. hyicus. The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6. shET activity in the culture filtrate from TY broth inoculated with 107 CFU of S. hyicus per ml was higher than that in TY broth inoculated with 106 and 108 CFU of bacteria per ml. When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions. shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor. shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask. shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose. Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 × 109 CFU of S. hyicus strain P‐1 into 300 ml of TY broth in a 2,000‐ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min. Then shET activity of the culture filtrate under appropriate culture conditions was measured after various incubation periods. shET activity was detected 6 hr after inoculation, reached the maximum (253 exfoliative unit/0.1 ml) at 16 hr and decreased between 20 and 48 hr. Thus, the optimum incubation period was determined to be 16 hr. Then the optimum concentration of ammonium sulfate for isolation of shET from the culture filtrate under appropriate culture conditions was examined. The greatest shET activity was obtained from the fraction salted out with 90% saturated ammonium sulfate. Thus, the optimum concentration of ammonium sulfate for the isolation of shET was determined to be 90% saturation.

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Purification of exfoliative toxin produced by Staphylococcus hyicus and its antigenicity

Cited by 30 publications

References 13 publications

Differentiation and distribution of three types of exfoliative toxin produced by Staphylococcus hyicus from pigs with exudative epidermitis

Differentiation and distribution of three types of exfoliative toxin produced by Staphylococcus hyicus from pigs with exudative epidermitis

Studies on the effect of divalent metal ions on exfoliative toxins from Staphylococcus hyicus: indications of ExhA and ExhB being metalloproteins

Optimum Culture Conditions for Production of Exfoliative Toxin by Staphylococcus hyicus

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