Lipid-binding allergens from Dermatophagoides pteronyssinus mites extract isolated by liposomes
© Picolo et al.; licensee BioMed Central Ltd. 2014
Published: 1 October 2014
House dust mites (HDM), including Dermatophagoides pteronyssinus (Dpt) and Dermatophagoides farinae (Df), are one of the commonest aeroallergens worldwide eliciting allergic manifestations . Allergenic phenomenon is tightly associated both with the mites themselves and with ligands derived from mite-associated bacterial/fungal products. Some Dpt allergens belong of lipid-binding proteins, including Der p 2 (protein with myeloid differentiation protein 2 (MD-2) related lipid recognition domain), Der p 7 (structurally homologous to lipid binding protein family) and Der p 13 (lipid transporter molecule) . Thus, in this study we aimed to evaluate the ability of liposomes to adsorb lipid-binding proteins from Dermatophagoites pteronyssinus mites extract.
Liposomes were prepared by ethanolic injection using dipalmitoyl phosphatidylcholine/cholesterol or dipalmitoyl phosphatidylethanolamine/ cholesterol diluted in ethanol. To Poll-down assays, Dpt allergens were incubate with liposome preparations and than washed three times with PBS solution. The adsorbed proteins on liposome surface were removed by SDS treatment and then analyzed by SDS-PAGE. Additionally, the lipid-binding proteins were analyzed by ELISA to evaluate the immunoreativity of Dpt-specific IgE and IgG1 antibodies .
Results and conclusions
Several proteins ranging from 21 to 205 kDa were enriched in poll-down assays, including a polypeptide with high molecular weight (>205 kDa). In addition, Dpt-adsorbed on liposomes were reactive to IgE and IgG1 antibodies from allergic patients analyzed by ELISA. Further analysis using mass spectrometry will be conducted to identify the proteins adsorbed on liposome surface. Liposomes might be used to produce enriched fractions of lipid-binding proteins from Dermatophagoides pteronyssinus for further studies in allergic diseases.
- Gandhi VD, Davidson C, Asaduzzaman M, Nahirney D, Vliagoftis H: House dust mite interaction with airway epithelium: Role in allergic airway inflammation. 2013, 13 (3): 262-270.Google Scholar
- Gregory LG, Lloyd CM: Orchestrating house dust mite-associated allergy in the lung. Trends in Immunology. 2011, 32 (9): 402-411. 10.1016/j.it.2011.06.006.PubMed CentralView ArticlePubMedGoogle Scholar
- Pereira EA, Silva DA, Cunha-Júnior JP, Almeida KC, Alves R, Sung SJ, Taketomi EA: IgE, IgG1 and IgG4 antibody responses to Blomia tropicalis in atopic pacientes. Allergy. 2005, 60 (3): 401-406. 10.1111/j.1398-9995.2005.00738.x.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.