Dataset Information

Characterization of arginine kinase, anovel allergen of dermatophagoides farinae (Der f 20).

ABSTRACT:

Objective

To characterize a novel allergen, the Dermatophagoides farinae-derived arginine kinase (Der f 20).

Methods

The protein of Der f 20 was synthesized by genetic engineering approaches. The allergenicity of Der f 20 was tested by enzyme-linked immunosorbent assay and an airway allergy mouse model.

Results

The Der f 20 gene was cloned andpresented in the Gene Bank with an accession number of AAP57094. The Der f 20 is an arginine kinase (AK), whichshowed a close relationship with D. pteronyssinus AK and Aleuroglyphusovatus AK. Western-blot and ELISA studies showed the IgE binding capacity of Der f 20 was 66.7% in the sera from 6 dust mite allergic patients. Immune inhibition assayresults showed the IgE cross-reactivity between Der f 20 and DME (Dust mite extract). Positive responses to Der f 20 were 41.2% as shown by skin prick tests in 17 DME-allergic patients. In vitro experimental results showed that Der f 20 induced Th2 cell differentiation and the expression of T cell Ig mucin domain molecule-4 (TIM4) in DCs. Conclusions; The Der f 20 protein is a novel subtype of thedust mite allergen.

SUBMITTER: Xing P

PROVIDER: S-EPMC4731678 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Characterization of arginine kinase, anovel allergen of dermatophagoides farinae (Der f 20).

Xing Peng P Yu Haiqiong H Li Meng M Xiao Xiaojun X Jiang Congli C Mo Lihua L Zhang Min M Yang Pingchang P Liu Zhigang Z

American journal of translational research 20151215 12

<h4>Objective</h4>To characterize a novel allergen, the Dermatophagoides farinae-derived arginine kinase (Der f 20).<h4>Methods</h4>The protein of Der f 20 was synthesized by genetic engineering approaches. The allergenicity of Der f 20 was tested by enzyme-linked immunosorbent assay and an airway allergy mouse model.<h4>Results</h4>The Der f 20 gene was cloned andpresented in the Gene Bank with an accession number of AAP57094. The Der f 20 is an arginine kinase (AK), whichshowed a close relatio ...[more]

PMID: 26885278

Similar Datasets

Project description:Background House dust mites (HDMs), including Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) species, represent a major source of inhalant allergens that induce IgE-mediated anaphylactic reactions. HDM allergen identification is important to the diagnosis and treatment of allergic diseases. Here, we report the identification of a novel HDM allergen, which we suggest naming Der f 40, and its immunodominant IgE epitopes. Methods The recombinant protein Der f 40 was expressed using a pET prokaryotic expression system and purified with Ni-NTA resins. IgE binding activity was evaluated by IgE-western blot, dot-blot, and ELISA. Mast cell activation testing was performed to assess the cellular effects of IgE binding in mouse bone marrow derived mast cells (BMMCs) expressing human FcεRI. IgE binding assays were performed with truncated and hybrid Der f 40 protein molecules to find immunodominant IgE epitopes. Results A 106-amino acid (aa) recombinant Der f Group 40 protein (rDer f 40) was obtained (GenBank accession No. XP_046915420.1) as thiredoxin-like protein. Der f 40 was shown to bind IgE from HDM allergic serum in vitro (9.68%; 12/124 in IgE-ELISA), and shown to promote the release of β-hexosaminidase from BMMCs dose-dependently when administered with HDM allergic sera. The Der f Group 40 protein was named Der f 40 and listed in the World Health Organization and International Union of Immunological Societies (WHO/IUIS) Allergen Nomenclature Sub-committee. IgE binding assays with Der f 40-based truncated and hybrid proteins indicated that IgE binding epitopes are likely located in the C-terminal region and dependent on conformational structure. The 76–106-aa region of C-terminus was identified as an immunodominant IgE epitope of Der f 40. Conclusion A novel HDM allergen with robust IgE binding activity was identified and named Der f 40. An immunodominant IgE epitope of Der f 40 with conformational dependency was identified in the C-terminus (aa 76–106). These findings provide new information that may be useful in the development of diagnostic and therapeutic agents for HDM allergy.

Dataset Information

Characterization of arginine kinase, anovel allergen of dermatophagoides farinae (Der f 20).

Objective

Methods

Results

Publications

Characterization of arginine kinase, anovel allergen of dermatophagoides farinae (Der f 20).

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets