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Human IgE mAbs define variability in commercial Aspergillus extract allergen composition
Mark A. Wurth, … , R. Stokes Peebles Jr., Scott A. Smith
Mark A. Wurth, … , R. Stokes Peebles Jr., Scott A. Smith
Published October 18, 2018
Citation Information: JCI Insight. 2018;3(20):e123387. https://doi.org/10.1172/jci.insight.123387.
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Research Article Immunology Pulmonology

Human IgE mAbs define variability in commercial Aspergillus extract allergen composition

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Abstract

Sensitization to Aspergillus species is associated with allergic respiratory diseases. Allergen immunotherapy with nonstandardized Aspergillus extracts is commonly used as therapy in these patients. Unfortunately, no method exists to measure the relevant allergen protein content in diagnostic and therapeutic extracts. Thus, there is a critical need for Aspergillus extract standardization. We hypothesized that development of Aspergillus-specific human IgE mAbs would allow for the characterization of the relevant human allergenic epitopes among currently available commercial Aspergillus fumigatus extracts. Patients with allergic bronchopulmonary mycosis were recruited from Vanderbilt University Medical Center. IgE antibody–secreting B cells were grown and immortalized using human hybridoma techniques first described here. Twenty-six human Aspergillus-reactive IgE mAbs were used as capture and detection reagents to characterize the Aspergillus allergen content of commercial extracts. We found extreme variability in the specificity and quantity of their protein targets. Just 4 mAbs reacted with all available extracts, and only 1 of 4 extracts contained the major allergen Asp f 1. This degree of variability will almost certainly affect the efficacy of these reagents when used in diagnosis and treatment. Human IgE mAbs represent an innovative tool for the evaluation of relevant human allergenic epitopes, which may assist in future development and long-term standardization of mold extracts.

Authors

Mark A. Wurth, Azadeh Hadadianpour, Dennis J. Horvath, Jacob Daniel, Olivia Bogdan, Kasia Goleniewska, Anna Pomés, Robert G. Hamilton, R. Stokes Peebles Jr., Scott A. Smith

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Figure 1

Identification of Asp f 1 in commercial extracts using human IgE mAbs.

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Identification of Asp f 1 in commercial extracts using human IgE mAbs.
(...
(A) Indirect fluorescence ELISA analysis of allergenic extracts. (B) Western blot (WB) analysis of allergenic extracts. Analyses were performed on commercially manufactured allergenic extracts from GREER, HS, ALKB, and AlrgyL using human IgE mAbs to the major allergen Asp f 1. Box-and-whisker plots show the median, first and fourth quartiles, and minimum and maximum values (whiskers).

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