Abstract
The house dust mite (HDM) Dermatophagoides pteronyssinus is an important risk factor for asthma and rhinitis. Allergen specific immunotherapy that is based on recombinant proteins has been proposed for the safer and more efficient treatment of allergic diseases. The aim of this study was to design and obtain a hybrid protein (DPx4) containing antigenic regions of allergens Der p 1, Der p 2, Der p 7, and Der p 10 from this mite. DPx4 was produced in Escherichia coli and its folding was determined by circular dichroism. Non-denaturing dot-blot, ELISA, basophil activation test, dot blot with monoclonal antibodies, ELISA inhibition, and cysteine protease activity assays were performed. Mice that were immunized with DPx4 were also analyzed. We found that DPx4 had no cysteine protease activity and it showed significantly lower IgE reactivity than Der p 1, Der p 2, and D. pteronyssinus extract. DPx4 induced lower basophil activation than Der p 2 and the allergen extract. Immunized mice produced IgG antibodies that inhibited the binding of allergic patient’s IgE to the allergen extract and induced comparatively higher levels of IL-10 than the extract in peripheral blood mononuclear cells (PBMC) culture. These results suggest that DPx4 has immunological properties that are useful for the development of a mite allergy vaccine.
Highlights
IgE antibodies to normally innocuous environmental antigens, such as house dust mite (HDM), pollens, and animal dander, mediate respiratory allergic diseases
DPx4 was Obtained as a Partly Folded Protein The DPx4 protein was recovered from the inclusion bodies, after treatment with 8 M urea
The rationale behind the design of the hybrid protein was to obtain one molecule containing antigenic regions from several D. pteronyssinus allergens with the inclusion of at least the major allergens Der p 1 and Der p 2, plus other allergens that contribute to the sensitization in allergic individuals
Summary
IgE antibodies to normally innocuous environmental antigens, such as house dust mite (HDM), pollens, and animal dander, mediate respiratory allergic diseases. The mite species Dermatophagoides pteronyssinus is very common in these regions [1,2] Several allergens from this species have been characterized, showing different capabilities of sensitization in atopic individuals [1,3]. Allergen specific immunotherapy (SIT) with whole allergen extract is the only disease that modifies treatment of allergy [4,5]. This approach has some disadvantages, such as great variation in composition, missing of important allergens, and the inclusion of non-relevant molecules [6]
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