Der p 5 allergen from house dust mite: first epitope mapping of rabbit IgG blocking antibodies

Lahiani, Sadjia; Dumez, Marie-Eve; Bitam, Idir; Galleni, Moreno

doi:10.1016/j.nmni.2018.11.009

Cited by 4 publications

(5 citation statements)

References 34 publications

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“…Of the 38 HDM allergen groups identified, allergens from 32 groups and 23 groups have been identified for Der f and Der p, respectively, and catalogued in the World Health Organization and International Union of Immunological Societies (WHO/IUIS) allergen database (http://www.allergen.org ). The most clinically important HDM allergens identified thus far have been group 1, group 2, and group 23 allergens [9 ], with recent evidence suggesting that HDM group 24 allergens may also be clinically important [10 ]. Previously, we found that a group 24 Der f allergen, namely Der f 24, was a ubiquinol cytochrome C reductase binding (UQCRB) protein homolog [10 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of immunodominant IgE binding epitopes of Der p 24, a major allergen of Dermatophagoides pteronyssinus

Cai

Chen

Zhang

et al. 2019

Clin Transl Allergy

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Background The identification of house dust mite (HDM) allergens and epitopes is important for allergy diagnosis and treatment. We sought to identify the Dermatophagoides pteronyssinus group 24 allergen (Der p 24) and to identify its immunodominant IgE epitope(s). Methods Der p 24 cDNA was cloned and expressed in a pET expression system. The IgE binding activity of purified recombinant (r)Der p 24 was evaluated by western blotting. Truncated Der p 24 proteins and overlapping synthetic polypeptides were subjected to IgE binding assays. Balb/c mice were immunized to investigate IgE epitope induction of IgE production. IgE binding of the 32 N-terminal residues of Der p 24 was compared to other Der p epitopes in enzyme-linked immunosorbent assays and dot blot assays. Human skin prick tests (SPTs) were performed. Results We cloned and expressed Der p 24 cDNA (GenBank accession no. KP893174.1). HDM allergic sera bound rDer p 24 in vitro and 5/10 HDM allergic patients (50%) had positive SPT reactions to rDer p 24. The immunodominant IgE epitope of Der p 24 was localized to the N-terminal 32-residue region, which produced a high specific IgE antibody titer in vivo and promoted mast cell β-hexosaminidase release. The IgE binding activity this N-terminal epitope of Der p 24 was stronger than that of Der p 1 or Der p 2 IgE epitopes. Conclusions We identified Der p 24 as a major HDM allergen with strong IgE binding activity via an immunodominant IgE epitope in the N-terminal 32-residue region, which triggers IgE production in vivo. The identified Der p 24 epitope may support HDM allergy diagnosis and treatment.

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Section: Introductionmentioning

confidence: 99%

Identification of immunodominant IgE binding epitopes of Der p 24, a major allergen of Dermatophagoides pteronyssinus

Cai

Chen

Zhang

et al. 2019

Clin Transl Allergy

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“…Bet v 1 dimerization provides two IgE binding sites that are required for cross-linking of IgE in effector cells and BCRs in B lymphocytes [56,60]. Interestingly, all these aforementioned dimeric allergens are known lipids carriers, as is rDer p 5 [16,19,26]. However, the investigation of IgE binding of a monomeric rDer p 5 in comparison with our oligomerized one would still be warranted, and we still believe that our IgE frequency was a relevant result.…”

Section: Discussionmentioning

confidence: 77%

“…Additionally, the number of monoclonal antibodies available for use in affinity chromatography is currently scarce, especially in the case of certain allergic sources [48,49]. For this reason, most research groups tend to heterologously express allergens and characterize them in vitro first [16,19,26]. With a few exceptions, most recombinant allergens displayed identical or similar allergenic activity and IgE binding capacity [43,45,50].…”

Section: Discussionmentioning

confidence: 99%

“…Considering previous reports on Der p 5 dimerization behavior [19,25,26,38], an initial in silico assessment of aggregating domains was performed using PASTA 2.0 [39]. Similarly to a previous analysis by our group [40], the amino acid sequence of shortened Der p 5 was uploaded to the PASTA 2.0 server, choosing the 100 top pairing energies and an energy threshold of 1.192 kcal/mol.…”

Section: In Silico and In Vitro Caracterization Of Rder P 5 Structura...mentioning

confidence: 99%

“…In addition, both mite groups are proteins exclusive of allergenic mites [8], and they are structurally similar [23,24]. These proteins present a predominantly α-helical structure, with the presence of a flexible N-terminal region and antiparallel α-helixes that, eventually, can polymerize creating a hydrophobic cavity [25,26]. 1 In bold countries with a tropical climate; only articles published up to 2007 are listed.…”

Section: Introductionmentioning

confidence: 99%

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Removal of N-Terminal Peptide Impacts Structural Aspects of an IgE-Reactive Recombinant Der p 5

Vieira,

Silva,

Silveira

et al. 2023

Allergies

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(1) Background: Modification of the structural elements of allergens is widely used in the field of allergies. The goal of the present research was to express, purify, and characterize the shortened recombinant group 5 allergen of Dermatophagoides pteronyssinus (rDer p 5). (2) Methods: rDer p 5 storage stability and aggregation capacity were explored through in silico analysis, dynamic light scattering (DLS), and SDS-PAGE. Serum IgE reactivity and cytokine amount were investigated in sera or cell culture supernatants through ELISA, MULTIPLEX®, and Western blot analysis using sera from sensitized humans from Brazil, Colombia, and Ecuador. (3) Results: Dimeric rDer p 5 was detected through native PAGE, and this result was confirmed by data from DLS. The protein was thermically stable, as it did not degrade at 4 °C for 21 days. The shortened rDer p 5 was classified as a major IgE allergen in Brazil and Colombia, but minor in Ecuador. IL-13, IL-10, IL-1β, and IL-6 were significantly elevated in the sera of rDer p 5-reactive patients. The same cytokines plus IL-5 were more secreted by human cells upon rDer p 5 stimulation. (4) Conclusions: N-terminal peptide deletion led to a higher rDer p 5 folding stability, which, even though dimeric, was an IgE-reactive protein. Therefore, rDer p 5 could be used for molecular diagnostic applications or as backbone for hypoallergen design.

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Identification of continuous immunoglobulin G epitopes of Dermatophagoides farinae allergens by peptide microarray immunoassay

et al. 2020

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Dermatophagoides farinae, as a common house dust mite species, is one of the main sources of allergens in the world. At present, Dermatophagoides farinae is found to contain more than 30 groups of allergens. These allergens are used for allergen‐specific immunotherapy (AIT) of allergic diseases. During the AIT process, immunoglobulin G (IgG) antibodies can block immunoglobulin E (IgE) antibody‐induced allergic reactions in the human body. One of the mechanisms may be that IgG and IgE competitively bind to the same allergic protein, so it is necessary to explore the binding sites (epitopes) of IgG antibodies to allergens. In this study, peptide arrays were constructed to react with the serums from patients with allergic asthma to find the IgG epitopes of several allergens including major allergens (Der f 1, 2) and mid‐tier allergens (Der f 4, 5, and 7), and then verified by enzyme‐linked immunosorbent assay (ELISA) test. Relevant epitopic sequences were located on the tertiary structure of individual allergens, as reconstructed by homology modeling. One IgG epitope of Der f 1 (90–106aa, NVPSELDLRSLRTVTPI), five IgG epitopes of Der f 4 (61–77aa, ERYQPVSYDIHTRSGDE; 193–209aa, FRSDASTHQWPDDLRSI; 226–242aa, HPFIYHETIYYGGNGIN; 271–287aa, LRWLRNFGTEWGLVPSG; 352–368aa, NDWVGPPTDQHGNILSV), and one IgG epitope of Der f 5 (84–101aa, RYNVEIALKSNEILERDL) were identified. IgG epitopes of Der f 2, 7 were not found. There are overlaps between the IgG and IgE epitopes of Der f 1, 4, and 5. These findings not only reflect the practicality of peptide array and ELISA test in the allergen IgG epitope identification, but also provide more information for further understanding of the human immunological changes during AIT and the molecular mechanisms of IgG blocking IgE activity.

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Der p 5 allergen from house dust mite: first epitope mapping of rabbit IgG blocking antibodies

Cited by 4 publications

References 34 publications

Identification of immunodominant IgE binding epitopes of Der p 24, a major allergen of Dermatophagoides pteronyssinus

Identification of immunodominant IgE binding epitopes of Der p 24, a major allergen of Dermatophagoides pteronyssinus

Removal of N-Terminal Peptide Impacts Structural Aspects of an IgE-Reactive Recombinant Der p 5

Identification of continuous immunoglobulin G epitopes of Dermatophagoides farinae allergens by peptide microarray immunoassay

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