Expression of Alt a 1 allergen from Alternaria alternata in the yeast Yarrowia lipolytica.
Allergies affect almost 25% of the population in industrialized countries. Alternaria alternata is known to be a significant source of aeroallergens and sensitization to this mold is a risk factor for the development of wheezing, asthma, and atopic dermatitis. Diagnosis and treatment of allergies requires the production of large amounts of pure and well defined protein. Yarrowia lipolytica, a non-pathogenic ascomycete able to secrete high levels of enzymes that can grow in inexpensive substrates, has been considered a useful host for heterologous gene expression. In the present work, we have developed two vectors for expressing Alt a 1, the most relevant A. alternata allergen, in Y. lipolytica. One vector is autosomal and one is integrative. With both systems, rAlt a 1 was secreted into the culture medium. The immunological characteristics of the purified recombinant allergen were determined by IgE-blot using sera from 42 A. alternata-allergic patients. We have carried out ELISA-inhibition experiments using sera from four patients to compare the IgE-binding capacity of natural and recombinant allergens. Our results show that Y. lipolytica is able to produce a recombinant Alt a 1 which is immunochemically equivalent to the natural counterpart and could be used for immunotherapy and diagnostics.
Department of Microbiology and Genetics, CIETUS, IBSAL Universidad de Salamanca, Salamanca, Spain.
Matías
M
Morín
Juan A
JA
Asturias
Angel
A
Domínguez
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000528:Alternaria / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D001249:Asthma / Q000276:immunology / Q000382:microbiology; D016022:Case-Control Studies; D002648:Child; D003470:Culture Media / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology*; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008297:Male; D011485:Protein Binding; D011994:Recombinant Proteins / Q000276:immunology; D012221:Rhinitis, Allergic, Perennial / Q000276:immunology / Q000382:microbiology; D025062:Yarrowia / Q000235:genetics / Q000378:metabolism*; D055815:Young Adult
C096258:ALTA1 protein, Alternaria alternata; D000485:Allergens; D003470:Culture Media; D000939:Epitopes; D005656:Fungal Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E