Preliminary Description of Antigenic Components Characteristic of Stachybotrys chartarum

Raunio, P.; Kärkkäinen, Marja; Virtanen, Tuomas; Rautiainen, J. Mikko; Pasanen, Anna‐Liisa

doi:10.1006/enrs.2000.4111

Cited by 18 publications

(28 citation statements)

References 52 publications

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“…To study the human IgE reactivity to different fungal species, the sera of five exposed subjects who had shown an intense reaction to the 48-kD component of S. chartarum and their matched control sera were used. In the analyses, the dilution used for human sera was 1:5, for the S. chartarum -specific rabbit polyclonal antiserum [18]1:10,000, for the rabbit anti-human Ig (P0295, Dako, Denmark) 1:1,000 and for the peroxidase-conjugated swine anti-rabbit Ig (P0399, Dako) 1:10,000. Only the reactions clearly exceeding the intensity of the negative control (serum was replaced with a blocking agent, gelatine-PBS-Tween) were regarded as positive.…”

Section: Methodsmentioning

confidence: 99%

“…SDS-PAGE, electrophoretic transfer and immunoblotting analyses were carried out substantially as described previously [18]. In the SDS-PAGE analysis, 10% acrylamide gel was used under reducing conditions.…”

Section: Methodsmentioning

confidence: 99%

“…Fungal antigen crude extracts (CE) were prepared substantially as described earlier [18]. Briefly, Alternaria alternata [Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) 62006], Aspergillus versicolor [University of Kuopio (UKU) 3], Cladosporium cladosporioides (DSMZ 62121), Memnoniella echinata (Centraalbureau voor Schimmelcultures 304.54), Penicillium brevicompactum [American Type Culture Collection (ATCC) 58606], Phialophora rebens (National Public Health Institute RE27), Phoma sp.…”

Section: Methodsmentioning

confidence: 99%

“…The human IgE reactivity against the 34- and 52-kD components of S. chartarum was recently reported [17]. In our previous study [18], we characterized an antigen profile for S. chartarum and reported the influence of culture media and incubation time on the antigenic composition of S. chartarum extracts. We also described the role of carbohydrate moieties in the IgE-binding capacity of S. chartarum proteins.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, cross-reactivity between S. chartarum and ten other fungal extracts was demonstrated using rabbit IgG antibodies. Based on the results, a 50-kD component in 15% acrylamide gel (shown in the present study as a 48-kD component in 10% acrylamide gel) was proved to be the most characteristic immunologically reactive component of S. chartarum [18]. This component was tentatively shown to possess human IgE reactivity in immunoblotting analyses [19].…”

Section: Introductionmentioning

confidence: 99%

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Partial Amino Acid Sequence of a Cellulase-Like Component with IgE-Binding Properties from <i>Stachybotrys chartarum</i>

Kärkkäinen

Raunio²,

Rautiainen³

et al. 2004

Int Arch Allergy Immunol

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Background: The aim of this study was to characterize the amino acid sequence of a selected Stachybotrys chartarum component and to investigate human IgE reactivity against components of S. chartarum and nine other fungal species. Methods: Human IgE reactivity against S. chartarum and nine other fungal extracts was investigated by the immunoblotting method. For automated amino acid sequencing analyses, the S. chartarum extract was purified by ion exchange chromatography prior to in-gel alkylation and digestion with modified trypsin. Results: Human IgE reactivity was detected against eight components in the S. chartarum extract. Over 80% of the sera from the exposed subjects and less than 50% of the control sera recognized the 33-, 48- and 50-kD S. chartarum components. The human sera detected a 48- to 50-kD component from the extracts of eight fungal species. Nineteen peptide sequences were identified from the 48-kD component of S. chartarum. An analysis of the peptide sequences revealed homology with known fungal glycoside hydrolase enzymes (cellulases). Conclusions: The data showed human IgE reactivity against several S. chartarum components, including one at 48 kD. On the other hand, the human sera recognized 48- to 50-kD components from seven other fungal species, suggesting shared antigenic components (e.g. enolase) between the fungi. Thus, to our knowledge, this is the first antigen identified from S. chartarum.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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