Characterization of a hybrid protein designed with segments of allergens from Blomia tropicalis and Dermatophagoides pteronyssinus

Martínez, Dalgys; Cantillo, José Fernando; Herazo, Helber; Wortmann, Judith; Keller, Walter; Caraballo, Luis; Puerta, Leonardo

doi:10.1016/j.imlet.2018.01.012

Cited by 10 publications

(16 citation statements)

References 53 publications

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“…Similar to other hybrid molecule of 22.8 kDa that was developed by our group (MAVAC-BD-2) [43], DPx4 induced IgG blocking antibodies in immunized mice, showed comparatively lower human serum IgE reactivity and basophil activation than the purified allergens and mite extract. However, there are major differences between these molecules in allergen representation and antigenic composition; DPx4 contains antigenic segments of four allergens from only D. pteronyssinus , with a larger contribution from Der p 1, while MAVAC-BD-2 contains antigenic segments of four allergens from this mite species and three allergens from B. tropicalis , with the largest contribution of Der p 2 and Blo t 5.…”

Section: Discussionmentioning

confidence: 80%

“…The plasmid containing of DPx4 insert was transformed into competent E. coli BL21 (DE3) cells (Invitrogen Corporation, Carlsbad, CA, USA) by electroporation. The expression and purification was performed, as described elsewhere [43]. In brief, the culture was induced with 1 mM IPTG for 4 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

“…26 sera were randomly chosen from the allergic patients to compare the IgE reactivity to DPx4 with that of Der p 1, Der p 2, and D. pteronyssinus extract for non-denaturing dot blot experiments, as described previously [43]. Three sera from non-allergic individuals and buffer alone were used as the controls.…”

Section: Methodsmentioning

confidence: 99%

“…The bound IgE and IgG antibodies were detected by incubation with 100 μL of alkaline phosphatase-conjugate anti human IgE (Sigma A3525) or anti-IgG (Sigma A3187). The substrate p-nitrophenyl phosphate (Sigma) diluted in 10% diethanolamine was added for the development of reactivity, and OD was measured in an ELISA reader at 405 nm [43]. The quantification of IgE was performed by using the standard curve of human IgE (Merk Millipore, Billerica, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

“…DPx4-specific IgG1 and IgG2a in serum were determined by ELISA while using MaxiSorp plate wells (Nunc, Thermo Fisher) that were coated with 1 µg/mL of DPx4. In addition, we evaluated whether IgG antibodies induced by immunization reacted with mite extract by dot blot assay, using nitrocellulose membrane that was dotted with 5 µg of allergen extract, as described in [43]. An ELISA inhibition test was performed using MaxiSorp plate wells that were coated with 5 µg of D. pteronyssinus extract and serum pool from the immunized mice with DPx4 to test whether the IgG antibodies could inhibit the human IgE binding to D. pteronyssinus extract, as described previously [43].…”

Section: Methodsmentioning

confidence: 99%

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An Engineered Hybrid Protein from Dermatophagoides pteronyssinus Allergens Shows Hypoallergenicity

Martínez

Múnera

Cantillo

et al. 2019

IJMS

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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.

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Section: Discussionmentioning

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

An Engineered Hybrid Protein from Dermatophagoides pteronyssinus Allergens Shows Hypoallergenicity

Martínez

Múnera

Cantillo

et al. 2019

IJMS

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The allergenic activity and clinical impact of individual IgE-antibody binding molecules from indoor allergen sources

Caraballo

Valenta

Puerta

et al. 2020

World Allergy Organization Journal

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A large number of allergens have been discovered but we know little about their potential to induce inflammation (allergenic activity) and symptoms. Nowadays, the clinical importance of allergens is determined by the frequency and intensity of their IgE antibody binding (allergenicity). This is a rather limited parameter considering the development of experimental allergology in the last 20 years and the criteria that support personalized medicine. Now it is known that some allergens, in addition to their IgE antibody binding properties, can induce inflammation through non IgE mediated pathways, which can increase their allergenic activity. There are several ways to evaluate the allergenic activity, among them the provocation tests, the demonstration of non-IgE mediated pathways of inflammation, case control studies of IgE-binding frequencies, and animal models of respiratory allergy. In this review we have explored the current status of basic and clinical research on allergenic activity of indoor allergens and confirm that, for most of them, this important property has not been investigated. However, during recent years important advances have been made in the field, and we conclude that for at least the following, allergenic activity has been demonstrated:

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Efficacy and safety of subcutaneous immunotherapy with a mixture of glutaraldehyde-modified extracts of Dermatophagoides pteronyssinus, Dermatophagoides farinae, and Blomia tropicalis

Cardona-Villa

Uribe-Garcia

Calvo-Betancur

et al. 2022

World Allergy Organization Journal

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Characterization of a hybrid protein designed with segments of allergens from Blomia tropicalis and Dermatophagoides pteronyssinus

Cited by 10 publications

References 53 publications

An Engineered Hybrid Protein from Dermatophagoides pteronyssinus Allergens Shows Hypoallergenicity

An Engineered Hybrid Protein from Dermatophagoides pteronyssinus Allergens Shows Hypoallergenicity

The allergenic activity and clinical impact of individual IgE-antibody binding molecules from indoor allergen sources

Efficacy and safety of subcutaneous immunotherapy with a mixture of glutaraldehyde-modified extracts of Dermatophagoides pteronyssinus, Dermatophagoides farinae, and Blomia tropicalis

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