Purification and characterisation of the dimeric group 12 allergen from Blomia tropicalis heterologously expressed by Escherichia coli Top10F´

Silva, Eduardo Santos da; Pacheco, Luis Gustavo Carvalho; Fernandes, Antônio Márcio Santana; Asam, Claudia; Silveira, Elisânia Fontes; Pinheiro, Carina S.; Alcantara-Neves, Neuza Maria

doi:10.1007/s11033-021-06361-6

Cited by 3 publications

(9 citation statements)

References 44 publications

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“…In addition, the positive correlations between sIgE measured by ImmunoCAP® to B. tropicalis and sIgE to recombinant proteins indicate that the amount of these allergens in the tested diagnostic tools was enough to indicate sensitization to this tropical mite. Nevertheless, our own extract displayed stronger correlations with the sIgE against the recombinant allergens, underscoring not only the quality of our extract, but also the need to continue the discussion on the importance of quality control of allergenic extract used for allergy diagnosis 10,21,25 …”

Section: Discussionmentioning

confidence: 93%

“…These results lead to the notion that both allergens should be included in underscoring not only the quality of our extract, but also the need to continue the discussion on the importance of quality control of allergenic extract used for allergy diagnosis. 10,21,25 The importance of these two B. tropicalis allergens was reinforced by the significant reduction in IgE binding of BTH2, an outcome that did not occur with BTH1. Again, the data emphasize the superior hypoallergenic aspect of BTH2, suggesting as well that this vaccine candidate can be used in other tropical regions of the globe to induce tolerance against its parental allergens.…”

Section: Discussionmentioning

confidence: 99%

“…Human IgE antibody reactivity to the four recombinant proteins and BtE was measured by indirect ELISA, as described. 18,25…”

Section: Human Ige-binding Assaymentioning

confidence: 99%

“…One day following the last challenge, the animals were euthanized with intraperitoneal injections of xylazine and ketamine (40 Several analyses regarding human IgE binding were performed for all three countries. 18,22,25 For PBMC-related results, given that the distribution was nonparametric, the Wilcoxon test was applied to verify statistical differences between stimulated-and non-stimulated PBMCs. 26 To compare the cytokine production of the two groups (allergic and non-allergic) for each stimulus, Mann-Whitney's test was used for the analyses.…”

Section: Evaluation Of Bth2 Treatment Capacity In Mice Sensitized Wit...mentioning

confidence: 99%

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The hybrid protein BTH2 suppresses allergic airway inflammation in a murine model of HDM‐specific immunotherapy

Silva

Santana

Silveira

et al. 2023

Clin Experimental Allergy

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BackgroundAllergen‐specific immunotherapy (AIT) is the only disease‐modifying treatment approach to change disease‐causing allergens. Hypoallergenic derivatives show promise as potential therapeutics, amongst which BTH2 was designed to induce tolerance against Blomia tropicalis allergy. Our aim was to investigate the hypoallergenicity and immunoregulatory activity of BTH2 in vitro and its therapeutic potential in a mouse model of AIT.MethodsRecombinant Blo t 5 and Blo t 21 allergens and their hybrid derivatives (BTH1 and BTH2) were expressed and purified. IgE binding capacity was tested by ELISA using sera from Brazilian, Colombian, and Ecuadorian subjects. Secretion of cytokines in supernatants from human cell cultures was measured following stimulation with the four recombinants and controls. The capacity of BTH2 to ameliorate allergic airway inflammation induced by B. tropicalis extract was evaluated in a murine model of AIT.ResultsrBlo t 5 and rBlo t 21 were identified as major allergens in Latin American patients, and BTH2 had the lowest IgE binding. In vitro stimulation of human cells induced greater levels of IL‐10 and IFN‐γ and reduced the secretion of Th2 cytokines. BTH2 ameliorated allergic airway inflammation in B. tropicalis‐challenged A/J mice, as evidenced by the histopathological and humoral biomarkers: decreased Th2 cytokines and cellular infiltration (especially eosinophils), lower activity of eosinophil peroxidase, an increase in IgG blocking antibodies and strong reduction of mucus production by goblet cells.ConclusionsOur study shows that BTH2 represents a promising candidate for the treatment of B. tropicalis allergy with hypoallergenic, immune regulatory and therapeutic properties. Further pre‐clinical studies are required in murine models of chronic asthma to further address the efficacy and safety of BTH2 as a vaccine against B. tropicalis‐induced allergy.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

“…Human IgE antibody reactivity to the four recombinant proteins and BtE was measured by indirect ELISA, as described. 18,25…”

Section: Human Ige-binding Assaymentioning

confidence: 99%

Section: Evaluation Of Bth2 Treatment Capacity In Mice Sensitized Wit...mentioning

confidence: 99%

See 2 more Smart Citations

The hybrid protein BTH2 suppresses allergic airway inflammation in a murine model of HDM‐specific immunotherapy

Silva

Santana

Silveira

et al. 2023

Clin Experimental Allergy

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“…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. For stability studies, the purified recombinant allergen was stored in NaP buffer at 4 • C (0 days to 21 days), as well as at RT (0 to 48 h).…”

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

confidence: 99%

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 rBlo t 41 with a chitin-binding type-2 domain: A novel major allergen from Blomia tropicalis

Luo,

Zhang,

Zheng

et al. 2024

International Journal of Biological Macromolecules

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Purification and characterisation of the dimeric group 12 allergen from Blomia tropicalis heterologously expressed by Escherichia coli Top10F´

Cited by 3 publications

References 44 publications

The hybrid protein BTH2 suppresses allergic airway inflammation in a murine model of HDM‐specific immunotherapy

The hybrid protein BTH2 suppresses allergic airway inflammation in a murine model of HDM‐specific immunotherapy

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

Identification of rBlo t 41 with a chitin-binding type-2 domain: A novel major allergen from Blomia tropicalis

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