Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it>
<p>Abstract</p> <p>Background</p> <p>Yeast surface display is a technique, where the proteins of interest are expressed as fusions with yeast surface proteins and thus remain attached to the yeast cell wall after expression. Our purpose was to study whether allergens ex...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2010-09-01
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| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/9/1/74 |
| _version_ | 1818565905291935744 |
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| author | Poulsen Lars K Søndergaard Ib Jensen Bettina M Borodina Irina |
| author_facet | Poulsen Lars K Søndergaard Ib Jensen Bettina M Borodina Irina |
| author_sort | Poulsen Lars K |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Yeast surface display is a technique, where the proteins of interest are expressed as fusions with yeast surface proteins and thus remain attached to the yeast cell wall after expression. Our purpose was to study whether allergens expressed on the cell surface of baker's yeast <it>Saccharomyces cerevisiae </it>preserve their native allergenic properties and whether the yeast native surface glycoproteins interfere with IgE binding. We chose to use the major allergens from the common wasp <it>Vespula vulgaris </it>venom: phospholipase A1, hyaluronidase and antigen 5 as the model.</p> <p>Results</p> <p>The proteins were expressed on the surface as fusions with a-agglutinin complex protein AGA2. The expression was confirmed by fluorescent cytometry (FACS) after staining the cells with antibody against a C-tag attached to the C-terminal end of the allergens. Phospholipase A1 and hyaluronidase retained their enzymatic activities. Phospholipase A1 severely inhibited the growth of the yeast cells. Antigen 5 - expressing yeast cells bound IgE antibodies from wasp venom allergic patient sera but not from control sera as demonstrated by FACS. Moreover, antigen 5 - expressing yeast cells were capable of mediating allergen-specific histamine release from human basophils.</p> <p>Conclusions</p> <p>All the three major wasp venom allergens were expressed on the yeast surface. A high-level expression, which was observed only for antigen 5, was needed for detection of IgE binding by FACS and for induction of histamine release. The non-modified <it>S. cerevisiae </it>cells did not cause any unspecific reaction in FACS or histamine release assay despite the expression of high-mannose oligosaccharides.</p> <p>In perspective the yeast surface display may be used for allergen discovery from cDNA libraries and possibly for sublingual immunotherapy as the cells can serve as good adjuvant and can be produced in large amounts at a low price.</p> |
| first_indexed | 2024-12-14T01:46:51Z |
| format | Article |
| id | doaj.art-175ce2cbc1ae4733a2afb16d6b4b55a0 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-14T01:46:51Z |
| publishDate | 2010-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-175ce2cbc1ae4733a2afb16d6b4b55a02022-12-21T23:21:31ZengBMCMicrobial Cell Factories1475-28592010-09-01917410.1186/1475-2859-9-74Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it>Poulsen Lars KSøndergaard IbJensen Bettina MBorodina Irina<p>Abstract</p> <p>Background</p> <p>Yeast surface display is a technique, where the proteins of interest are expressed as fusions with yeast surface proteins and thus remain attached to the yeast cell wall after expression. Our purpose was to study whether allergens expressed on the cell surface of baker's yeast <it>Saccharomyces cerevisiae </it>preserve their native allergenic properties and whether the yeast native surface glycoproteins interfere with IgE binding. We chose to use the major allergens from the common wasp <it>Vespula vulgaris </it>venom: phospholipase A1, hyaluronidase and antigen 5 as the model.</p> <p>Results</p> <p>The proteins were expressed on the surface as fusions with a-agglutinin complex protein AGA2. The expression was confirmed by fluorescent cytometry (FACS) after staining the cells with antibody against a C-tag attached to the C-terminal end of the allergens. Phospholipase A1 and hyaluronidase retained their enzymatic activities. Phospholipase A1 severely inhibited the growth of the yeast cells. Antigen 5 - expressing yeast cells bound IgE antibodies from wasp venom allergic patient sera but not from control sera as demonstrated by FACS. Moreover, antigen 5 - expressing yeast cells were capable of mediating allergen-specific histamine release from human basophils.</p> <p>Conclusions</p> <p>All the three major wasp venom allergens were expressed on the yeast surface. A high-level expression, which was observed only for antigen 5, was needed for detection of IgE binding by FACS and for induction of histamine release. The non-modified <it>S. cerevisiae </it>cells did not cause any unspecific reaction in FACS or histamine release assay despite the expression of high-mannose oligosaccharides.</p> <p>In perspective the yeast surface display may be used for allergen discovery from cDNA libraries and possibly for sublingual immunotherapy as the cells can serve as good adjuvant and can be produced in large amounts at a low price.</p>http://www.microbialcellfactories.com/content/9/1/74 |
| spellingShingle | Poulsen Lars K Søndergaard Ib Jensen Bettina M Borodina Irina Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> Microbial Cell Factories |
| title | Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> |
| title_full | Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> |
| title_fullStr | Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> |
| title_full_unstemmed | Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> |
| title_short | Display of wasp venom allergens on the cell surface of <it>Saccharomyces cerevisiae</it> |
| title_sort | display of wasp venom allergens on the cell surface of it saccharomyces cerevisiae it |
| url | http://www.microbialcellfactories.com/content/9/1/74 |
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