Immunoblot Analyses of the Serologic Response to Aspergillus fumigatus Antigens in Experimental Invasive Aspergillosis

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Abstract
Theantibody response to Aspergillus fumigatus proteins was studied by the immunoblot technique in a rabbit model of invasive aspergillosis. Components of an A. fumigatus mycelial homogenate unbound by concanavalin A-Sepharose 4B chromatography were fractionated using SDS-PAGE and transferred to nitrocellulose papers. The protein fraction was probed with serial sera obtained from immunosuppressed or nonimmunosuppressed rabbits inoculated intravenously with saline or gradedinocula of A. fumigatus conidia. Seroconversion against antigens of 41, 54,and 71 kDa was demonstrated in 7 (50%),3 (21%), and 3 (21%) of 14 infected animals that survived ⩾10 days. Antibodies against the three antigens were detected in 4 (12.5%), 19(59%), and 14(44%) of 32 rabbits before immunosuppression or infection. Two-dimensional immuno-blotting revealed that the 41-, 54-, and 71-kDa antigens were derived in denaturing conditions from a single component resolved in nondenaturing polyacrylamide gel electrophoresis.

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