Study of the specific activity of the granulated form of the mixed-allergen from house dust mites Dermatophagoides pteronyssinus and Dermatophagoides farinae for oral immunotherapy

Юрьевна²,

Khlgatian

et al. 2021

Reviews of domestic and foreign authors consider different approaches to understanding the formation of immunological and clinical tolerance induced by allergen-specific immunotherapy (ASIT). Despite the wide variety of theoretical research, the mechanism of the bodys immune systems response to ASIT remains unclear. The aim of this review is to analyze the current understanding of the mechanisms of formation of changes in the bodys reactivity in response to an allergen after ASIT. It is known that the type of response to the antigen is determined by its dose. In low-dose ASIT tolerance to the antigen is formed in the absence of inflammation, which is apparently associated with the activation of specific high-affinity receptors on cells of the immune system. High doses of allergen in ASIT probably lead to a rearrangement of cellular receptors, causing a decrease in their number by internalization or a weakening of their sensitivity to an excessive signal (desensitization). Due to a decrease in the number of receptors and / or their loss of sensitivity, the response to the antigen changes according to the principle of negative regulation, implemented at the level of receptor or postreceptor mechanisms. The formation of an anti-inflammatory cytokine response to antigen contributes to the differentiation of naive T cells into inducible regulatory T cells (iTreg). The suppressing effect of Treg on immune system cells affects Th effector cells, mast cells, basophils, eosinophils, B cells, and dendritic cells. The occurring immunological shifts form a new type of tolerant response to the allergen, namely, the change in the type of immunoglobulins from IgE to IgG and IgA and new phenotypes of T memory and B memory cells.

Modern ideas about mechanisms of allergen-specific immunotherapy

Юрьевна²,

Khlgatian

et al. 2021

Treatment forms of house dust mites allergens intended for allergen-specific immunological therapy

Vasilyeva

Berzhets²,

et al. 2021

The growth of allergic diseases dictates the need to develop new forms of therapeutic allergens with high immunogenic and low allergenic activity. For many years, our laboratory has been developing drugs for the diagnosis and treatment of house dust mites (HDM) allergies. The purpose of this study is to summarize the results of the five-year development of therapeutic preparations of HDM allergens. During this period, we obtained the following forms of therapeutic allergens: a granular sublingual dosage form of a mixed allergen from Dermatophagoides pteronyssinus (Der.p) and Dermatophagoides farinaе (Der.f) and a succinylated monomeric HDM allergoid Der.p. The physicochemical and immunobiological properties of the obtained preparations were studied by methods: electrophoresis in PAGE in the presence of SDS-sodium, micropoint immunoblot, ELISA, inhibition of the binding reaction of allergen-specific IgE in the sera of patients. The research results showed that the obtained preparations have a reduced allergenic and increased immunogenic activity in comparison with native extracts. The created forms of mite allergens can be further used to treat patients sensitized to HDM of the genus Dermatophagoides.

Actual questions of standardization of water-salt extracts of house dust mites mixed allergens intended for medicinal forms of sublingual immunotherapy

Berzhets

Нестеренко

et al. 2022

Background: Standardization of medical forms of allergens is the process of establishing and applying a unified system of quality indicators and control methods. Quality control of therapeutic forms of allergens is an essential condition for guaranteeing the clinical efficacy and safety of allergen-specific immunotherapy (ASIT). The technology for obtaining the granular form of the mixed allergen from house dust mites Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) for oral use was developed in the laboratory of allergens Mechnikov Research Institute of Vaccines and Sera. Aims: to determine the authenticity of the anphenolic water-salt extract of the mixed allergen from mites Der p and Der f, intended for the therapeutic form of the drug. Materials and methods: Standardization of the obtained allergen extract was carried out by protein nitrogen unit by the Nessler method. The protein concentration was determined by the Bradford method. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to study the presence of protein composition in mixed allergen. The pH value was determined potentiometrically. The comparison of the specific activity of the standard allergen and the experimental mixed allergen was carried out by the method of reverse immunoassay (REAST). Results: SDS-PAGE showed the preservation of the protein profile of the water-salt extract of the mixed allergen Der p and Der f, corresponding to the protein profile of allergens of house dust mites. There were no statistically significant differences in the optical density of the studied allergen (four drug series) and the standard allergen with reference serums containing specific IgE to Der p or Der f when reacting with a Class 3 reference serum in REAST(р0,05). Conclusions: The above in accordance with the State Pharmacopoeia of the Russian Federation XIV edition is a confirmation of the authenticity of the drug. Not phenolic extraction did not affect the main allergenic components of the mixed allergen Der p and Der f and allowed it to maintain a high degree of specificity.