Production profile of the major allergen Alt a 1 in Alternaria alternata cultures.
BACKGROUND
The fungus Alternaria is strongly associated with asthma, but the importance of fungal allergen products is frequently underestimated. The profile of allergen release from fungal material is poorly understood.


OBJECTIVE
To investigate expression of the major allergen of Alternaria alternata, Alt a 1, during its growth in culture conditions for allergen extract production.


METHODS
Allergen expression was examined by Alt a 1-specific 2-site monoclonal antibody enzyme-linked immunosorbent assay, immunoblotting, and potency assays. The release of Alt a 1 was studied by transmission electron microscopy in conjunction with immunogold staining by using antibodies with specificity for Alt a 1.


RESULTS
A maximum amount of Alt a 1 was obtained after 4.5 weeks of growing, and it was found predominantly in the spent culture medium. In the same way, total IgE binding activity showed 15-fold more activity in the spent culture medium than in the buffer-extractable antigen fraction. Immunogold electron microscopy provided evidence that Alt a 1 is released from spores and mycelia.


CONCLUSIONS
Alternaria alternata allergenic proteins were constantly released into the culture medium, where they accumulated. Alt a 1 was a good marker for checking optimal culture conditions for A alternata extract production intended for clinical use.
Ibarrola
Ignacio
I
Departamento de Investigación y Desarrollo, Bial-Arístegui, Bilbao, Spain.
Suárez-Cervera
María
M
Arilla
M Carmen
MC
Martínez
Alberto
A
Monteseirín
Javier
J
Conde
José
J
Asturias
Juan A
JA
D000528:Alternaria / Q000276:immunology* / Q000648:ultrastructure*; D000818:Animals; D000946:Antigens, Fungal / Q000096:biosynthesis* / Q000276:immunology; D015153:Blotting, Western; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007073:Immunoglobulin E / Q000276:immunology; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D013110:Sphingosine / Q000096:biosynthesis* / Q000276:immunology; D013172:Spores, Fungal / Q000276:immunology*
C047996:Alternaria alternata pathotoxin TA; D000946:Antigens, Fungal; D007073:Immunoglobulin E; D013110:Sphingosine