Identification and analysis of basidiospore allergens from puffballs.
Spore extracts from four common puffballs (Calvatia cyathiformis, Geaster saccatum, Pisolithus tinctorius, and Scleroderma areolatum) were fractionated by Sephadex G-75 gel filtration. Pooled fractions, analyzed by skin test or direct RAST, contained allergenic components over a molecular weight range of 10,500 to 20,000 daltons for all puffball spores studied except for S. areolatum (greater than 70,000 daltons). The Sephadex G-75 allergenic pool of C. cyathiformis was further fractionated by hydrophobic interaction chromatography. Fractions were successively eluted with 4 and 1 mol/L buffered saline solutions and water. The 1 mol/L fraction contained maximal allergenic activity as determined by RAST-inhibition studies. At least 15 Coomassie blue staining bands were identified after isoelectrofocusing, of which 7 were shown by immunoprinting to bind IgE from a serum pool of five patients with allergy. Our results demonstrate that a combination of gel filtration, hydrophobic interaction chromatography, RAST-inhibition, and immunoprinting studies can be used to fractionate, characterize, and standardize spore allergen extracts of puffballs for diagnostic and therapeutic purpose.
Ibanez
M D
MD
Department of Medicine, Tulane University School of Medicine, New Orleans, LA 70112.
Horner
W E
WE
Liengswangswong
V
V
Sastre
J
J
Lehrer
S B
SB
D000485:Allergens / Q000032:analysis*; D000946:Antigens, Fungal / Q000032:analysis*; D001487:Basidiomycota / Q000276:immunology*; D002850:Chromatography, Gel; D003911:Dextrans; D006801:Humans; D015151:Immunoblotting; D007525:Isoelectric Focusing; D011852:Radioallergosorbent Test; D012882:Skin Tests; D013172:Spores, Fungal / Q000276:immunology*
D000485:Allergens; D000946:Antigens, Fungal; D003911:Dextrans; C025614:sephadex