Purification of a major allergen, Asp f 2 binding to IgE in allergic bronchopulmonary aspergillosis, from culture filtrate of Aspergillus fumigatus.
BACKGROUND
Most cases of allergic bronchopulmonary aspergillosis (ABPA) are caused by the fungus Aspergillus fumigatus. Successful treatment of this disease depends on early diagnosis with the use of well-characterized and relevant antigens/allergens of the organism.


OBJECTIVE
The aim of this study was to purify and characterize relevant proteins from A. fumigatus that could be used in the reliable diagnosis of ABPA.


METHODS
Monoclonal antibodies were raised against A. fumigatus culture filtrate antigens. A Concanavalin A nonbinding protein fraction was purified with use of one of the monoclonal antibody immunoaffinity columns. The purified protein was analyzed on sodium dodecylsulfate-polyacrylamide gel electrophoresis gel and Western blots. The sensitivity and specificity of the purified protein were evaluated by RAST and ELISA with sera from 25 patients with ABPA, from 10 with allergic asthma, and from 10 normal control subjects.


RESULTS
The 37 kD Concanavalin A nonbinding protein reacted specifically with IgE antibodies in patients with ABPA. Among the 25 patients with ABPA studied, 96% had IgE antibody against the allergen, whereas none of the subjects with allergic asthma who had positive results on the skin prick test or normal control subjects had a reaction. Both RAST and ELISA results exhibited strong correlation with IgE binding. This allergen exhibited N-terminal sequence identity to a recombinant allergen Asp f 2.


CONCLUSIONS
A 37 kD protein with complete N-terminal homology to Asp f 2 is a major allergen of A. fumigatus that significantly reacts with IgE antibody in patients with ABPA, but does not elicit reaction in Aspergillus-sensitive subjects with asthma and normal control subjects.
Banerjee
B
B
Research Service, Veterans Affairs Medical Center, Milwaukee, Wis. 53295, USA.
Kurup
V P
VP
Greenberger
P A
PA
Hoffman
D R
DR
Nair
D S
DS
Fink
J N
JN
D000485:Allergens / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000918:Antibody Specificity; D000946:Antigens, Fungal / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D001229:Aspergillosis, Allergic Bronchopulmonary / Q000276:immunology*; D001232:Aspergillus fumigatus / Q000276:immunology* / Q000378:metabolism; D001666:Binding Sites, Antibody; D002478:Cells, Cultured; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D003208:Concanavalin A; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D011852:Radioallergosorbent Test
D000485:Allergens; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000939:Epitopes; D005656:Fungal Proteins; C106810:allergen Asp f 2, Aspergillus fumigatus; D003208:Concanavalin A; D007073:Immunoglobulin E