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Published on: May 30, 2020
Characterization of folded recombinant Der p 5, a potential diagnostic marker allergen for house dust mite allergy
Margit Weghofer1, Monika Grote, Yuliya Dall'Antonia
1Division of Immunopathology, Department of Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
Background:
Der p 5 was reported as an important allergen in Dermatophagoides pteronyssinus, which is particularly recognized by patients suffering from asthma. The aim of this study was to produce, by recombinant DNA technology, a folded Der p 5 allergen for diagnostic, therapeutic and preventive purposes.
Methods:
Der p 5-encoding cDNA was isolated from a lambda gt11 D. pteronyssinus expression cDNA library and expressed in Escherichia coli. rDer p 5 was purified to homogeneity and characterized by mass spectroscopy and circular dichroism. IgE reactivity was tested with sera from 117 mite-allergic patients and in a basophil histamine release experiment. Der p 5-specific rabbit IgG antibodies were produced for the ultrastructural localization of the allergen in mites by immunogold electron microscopy as well as for cross-reactivity studies.
Results:
rDer p 5 is a heat-stable protein with predominantly alpha-helical secondary structure which reacted with IgE from 31% of mite-allergic patients' sera and showed no relevant cross-reactivity to group 5 allergens from storage mites and tropical mites. rDer p 5-specific rabbit IgG antibodies inhibited mite-allergic patients' IgE binding to Der p 5 and allowed to localize the allergen in secretory granules of midgut epithelial cells of house dust mites.
Conclusions:
The described rDer p 5 molecule may be useful for diagnosis and immunotherapy of house dust mite-allergic patients.
