The Nature of Staphylococcal Beta Hemolysin: I. Mode of Action

Wiseman, G. M.; Caird, J. D.

doi:10.1139/m67-049

Cited by 35 publications

(14 citation statements)

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“…Calcium influx may be involved in the activation of Erk-type MAP kinases as observed in other cell types (24). Staphylococcal Hlb has sphingomyelinase activity (54). Hydrolysis of sphingomyelin from the outer leaflet of the plasma membrane of potential host cells results in the generation of phosphorylcholine and ceramides.…”

Section: Discussionmentioning

confidence: 99%

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Virulence factors ofStaphylococcus aureusinduce Erk-MAP kinase activation and c-Fos expression in S9 and 16HBE14o- human airway epithelial cells

Below¹,

Konkel²,

Zeeck³

et al. 2009

American Journal of Physiology-Lung Cellular and Molecular Phys

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Part of the innate defense of bronchial epithelia against bacterial colonization is regulated secretion of salt, water, and mucus as well as defensins and cytokines involving MAP kinase activation and alterations in early gene expression. We tested two different types of immortalized human airway epithelial cells (S9, 16HBE14o-) for activation of Erk-type MAP kinases and for expression of c-Fos on treatment with Staphylococcus aureus culture supernatants from the stationary growth phase [optical density (OD)(540 nm) = 10] or with recombinant S. aureus hemolysins A and B (Hla, Hlb). OD10 supernatants activated Erk-type MAP kinases and c-Fos expression in a concentration-dependent manner. Hla induced Erk-type kinase phosphorylation in S9 but not in 16HBE14o- cells. Hlb induced Erk activation in either cell type. Basal and stimulated levels of Erk-type MAP kinase phosphorylation were sensitive to the Mek1 inhibitor PD-98059, indicating that the bacterial products activated the entire signaling cascade that coregulates IL-8 induction and secretion. While c-Fos expression was enhanced by OD10 supernatants, Hla, and Hlb in S9 cells, 16HBE14o- cells responded to OD10 supernatant and Hlb but not to Hla. In S9 cells, PD-98059 suppressed c-Fos upregulation by OD10 supernatant, Hla, or Hlb, indicating that c-Fos expression requires activation of Erk-type MAP kinases. In 16HBE14o- cells, however, c-Fos expression by OD10 supernatant was sensitive to PD-98059, while that induced by Hlb was not. This indicates that ingredients of OD10 supernatants other than Hla or Hlb are activating Erk-type MAP kinases in 16HBE14o- cells and that other intracellular signaling systems apart from Erk-type MAP kinases contribute to Hlb-mediated regulation of c-Fos. Thus interaction of bacterial factors with airway epithelial cells may be highly cell type specific.

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Section: Discussionmentioning

confidence: 99%

“…Staphylococcal Hlb has sphingomyelinase activity (54). Hydrolysis of sphingomyelin from the outer leaflet of the plasma membrane of potential host cells results in the generation of phosphorylcholine and ceramides.…”

mentioning

confidence: 99%

Virulence factors ofStaphylococcus aureusinduce Erk-MAP kinase activation and c-Fos expression in S9 and 16HBE14o- human airway epithelial cells

Below¹,

Konkel²,

Zeeck³

et al. 2009

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Purified ,!I-lysin was used in one immunodiffusion study and was prepared by a modification of the method of Wiseman and Caird (1967) in which the haemolysin was passed through Sephadex G-75 equilibrated with Hallander's (1963) buffer. The active lysin was pooled when removed from the column, dialysed against distilled water for 48 hours, centrifuged and the supernatant fluid lyophilised.…”

Section: Methodsmentioning

confidence: 99%

Immunogenicity of the -Haemolysin of Staphylococcus Aureus

Fackrell

Wiseman

1974

Journal of Medical Microbiology

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PLATE XIXAT T E M P,T s by several investigators have failed to demonstrate the immunogenicity of the 6-haemolysin of StaphyZococcus aureus. Gladstone and Yoshida (1967) did not convincingly demonstrate that 8-lysin was immunogenic and they showed that inhibitors of its haemolytic activity were present in serum. Recent evidence suggests that these serum inhibitors are lipoproteins (Donahue, 1969;Kantor, Temples and Shaw, 1972).In this study, the immunogenicity of 8-lysin has been demonstrated by the use of purified antibody preparations. MATERIALS AND METHODSPreparation of 6-lysin The strain employed, methods of production and purification of 6-lysin were those of Caird and Wiseman (1970). Purified S-lysin was also kindly sent to us by Dr Arnold Kreger. Haemolysin titrations were performed as described by Wiseman and Caird (1972). Buffers were prepared according to Gomori (1955) and, where required, 0 . 0 1~ phosphate buffer at pH 7 was supplemented with 0.85% NaCl. Nitrogen content was assayed by the microKjeldahl technique as described by Markham (1942) and an estimate of protein content was derived from the assay value multiplied by 6.25.Preparation of u-, ,8-and 7-lysins u-and y-Haemolysins were prepared and purified as described elsewhere (Fackrell and Wiseman, 1974; Wiseman, Caird and Fackrell, 1975). Purified ,!I-lysin was used in one immunodiffusion study and was prepared by a modification of the method of Wiseman and Caird (1967) in which the haemolysin was passed through Sephadex G-75 equilibrated with Hallander's (1963) buffer. The active lysin was pooled when removed from the column, dialysed against distilled water for 48 hours, centrifuged and the supernatant fluid lyophilised.Serological procedures Immunodi'usion, immunoelectrophoresis and quantitative precipitin methods were those of Campbell et al. (1970). The blood agarose required in some immunodiffusion tests was prepared by the addition of 1.0 ml of packed, washed human group-0 RBC to 100 ml of a 1 % (w/v) solution of Difco agarose in phosphate-buffered saline held at 50°C.Production of antisera. Antibodies to the haemolysins were prepared in 2-month-old New Zealand white rabbits purchased from the Canadian Research Animal Farm, Bradford,

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“…α -toxin does not directly shed syndecan-1 ectodomains, but rather stimulates an endogenous mechanism which involves protein tyrosine kinases (e.g., Syk), but not protein kinase C and MAPK signaling pathways, that enhance the cleavage of syndecan-1 ectodomains by host cell metalloproteinase [100]. Staphylococcal β -toxin enhances syndecan-1 shedding by activating ceramide production in the alveolar epithelial cells and by implicating protein tyrosine kinases Syk and JAK2, Erk-type MAPKs, and metalloproteinase [115, 116]. …”

Section: Known Virulence Factors Of S Aureus and Their Novel Funcmentioning

confidence: 99%

Characterization of Virulence Factors ofStaphylococcus aureus: Novel Function of Known Virulence Factors That Are Implicated in Activation of Airway Epithelial Proinflammatory Response

Bień

Sokolova

Bożko

2011

Journal of Pathogens

154

124

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Airway epithelial cells play a major role in initiating inflammation in response to bacterial pathogens. S. aureus is an important pathogen associated with activation of diverse types of infection characterized by inflammation dominated by polymorphonuclear leukocytes. This bacterium frequently causes lung infection, which is attributed to virulence factors. Many of virulence determinants associated with S. aureus-mediated lung infection have been known for several years. In this paper, we discuss recent advances in our understanding of known virulence factors implicated in pneumonia. We anticipate that better understanding of novel functions of known virulence factors could open the way to regulate inflammatory reactions of the epithelium and to develop effective strategies to treat S. aureus-induced airway diseases.

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The Nature of Staphylococcal Beta Hemolysin: I. Mode of Action

Cited by 35 publications

References 10 publications

Virulence factors ofStaphylococcus aureusinduce Erk-MAP kinase activation and c-Fos expression in S9 and 16HBE14o- human airway epithelial cells

Virulence factors ofStaphylococcus aureusinduce Erk-MAP kinase activation and c-Fos expression in S9 and 16HBE14o- human airway epithelial cells

Immunogenicity of the -Haemolysin of Staphylococcus Aureus

Characterization of Virulence Factors ofStaphylococcus aureus: Novel Function of Known Virulence Factors That Are Implicated in Activation of Airway Epithelial Proinflammatory Response

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