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To increase the effectiveness and immunogenicity of modern vaccines, especially subunit ones, it is required to use adjuvants. Polysaccharides, due to their safety and biocompatibility, are desirable candidates for the creation of vaccine adjuvants. The aim of our study was to develop a method for obtaining beta-Glucans from the yeast Saccharomyces cerevisiae cell wall, and evaluate their adjuvant properties. The high purity and non-toxicity of the resulting preparation was achieved by using enzyme complexes of cellulase and protease in combination with ultrasound (22 kHz) at the purification stage. The developed scheme allows for the yield of beta-Glucans up to 2 g from 100 g of the biomass of wet cells. The adjuvant properties of beta-Glucans were studied in 50 male BALB/c mice, weighing 16–18 g. Immunization was performed twice, with a 14-day interval, intramuscularly, 200 μl per animal. The recombinant receptor-binding domain (RBD) of the surface S protein of the SARS-CoV-2 virus (Wuhan-Hu-1 and B.1.617.2 (Delta)) was used as an antigen, at a dose of 50 μg per animal. A positive control group was administered with the antigen combined with aluminum hydroxide. As a negative control, mice injected with the saline solution were used. The titers of specific antibodies in the blood sera were determined by ELISA assays. RBD (Wuhan-Hu-1 and Delta), and S protein (Wuhan-Hu-1, Delta and Omicron) were used as antigens. The titers of virus-neutralizing antibodies were measured in neutralization tests using SARS-CoV-2 virus strains Wuhan-Hu-1, Delta (B.1.617.2) and Omicron (B.1.1.529). The results of the study have shown that beta-Glucans have the ability to enhance the production of specific and virus-neutralizing antibodies in mice immunized with RBD. The titers of specific and virus neutralizing antibodies are comparable to their levels in the group immunized with RBD and Al(OH)₃. It has been found in the experiments in white outbred ICR mice that the preparation belongs to practically non-toxic substances. Therefore, it can be concluded that the use of beta-Glucans could become a preferable alternative to the conventional adjuvants based on aluminum salts, being biocompatible, biodegradable and non-toxic substances of low labor-intensive production.
To increase the effectiveness and immunogenicity of modern vaccines, especially subunit ones, it is required to use adjuvants. Polysaccharides, due to their safety and biocompatibility, are desirable candidates for the creation of vaccine adjuvants. The aim of our study was to develop a method for obtaining beta-Glucans from the yeast Saccharomyces cerevisiae cell wall, and evaluate their adjuvant properties. The high purity and non-toxicity of the resulting preparation was achieved by using enzyme complexes of cellulase and protease in combination with ultrasound (22 kHz) at the purification stage. The developed scheme allows for the yield of beta-Glucans up to 2 g from 100 g of the biomass of wet cells. The adjuvant properties of beta-Glucans were studied in 50 male BALB/c mice, weighing 16–18 g. Immunization was performed twice, with a 14-day interval, intramuscularly, 200 μl per animal. The recombinant receptor-binding domain (RBD) of the surface S protein of the SARS-CoV-2 virus (Wuhan-Hu-1 and B.1.617.2 (Delta)) was used as an antigen, at a dose of 50 μg per animal. A positive control group was administered with the antigen combined with aluminum hydroxide. As a negative control, mice injected with the saline solution were used. The titers of specific antibodies in the blood sera were determined by ELISA assays. RBD (Wuhan-Hu-1 and Delta), and S protein (Wuhan-Hu-1, Delta and Omicron) were used as antigens. The titers of virus-neutralizing antibodies were measured in neutralization tests using SARS-CoV-2 virus strains Wuhan-Hu-1, Delta (B.1.617.2) and Omicron (B.1.1.529). The results of the study have shown that beta-Glucans have the ability to enhance the production of specific and virus-neutralizing antibodies in mice immunized with RBD. The titers of specific and virus neutralizing antibodies are comparable to their levels in the group immunized with RBD and Al(OH)₃. It has been found in the experiments in white outbred ICR mice that the preparation belongs to practically non-toxic substances. Therefore, it can be concluded that the use of beta-Glucans could become a preferable alternative to the conventional adjuvants based on aluminum salts, being biocompatible, biodegradable and non-toxic substances of low labor-intensive production.
Searching for a preparation that would meet all the requirements for modern adjuvants remains a matter of critical importance for specific immunoprophylaxis. Much information is available now on chitosan positive effect, including its effect on the immune response. The article provides results of the preclinical tests for different affordable chitosan-based products. For the test purposes, we took the following three products manufactured by LLC Bioprogress (Shchelkovo, Russia): water-soluble chitosan (succinate) – 2% solution edible chitosan (water-soluble) – 2% solution; edible chitosan (acid-soluble) – 2% solution, as well as antirabies vaccine RABIKOV manufactured by Shchelkovo Biocombinat (Russia). Immunogenic properties of chitosan-based products were tested in 85–100-day-old female white laboratory mice weighing 21–35 g. The animals were divided into 37 groups (6 mice in each group). Chitosan-based products were administered subcutaneously or intramuscularly, either together with the anti-rabies vaccine or without it. Animals from the control groups received either saline solution or the vaccine only. There was also a group of intact animals. The experiment demonstrated that the water-soluble chitosan (succinate) administered subcutaneously, acid-soluble edible chitosan (at a concentration of 1:64 and more), and water-soluble edible chitosan (at a concentration of 1:108) administered subcutaneously and intramuscularly increase the level of post-vaccination anti-rabies antibodies. Thus, the tested chitosan-based products do not have any negative impact on the laboratory animals and have immunogenic properties.
The purpose of this review is to analyze scientific data on the adjuvant properties of substances of various origin and chemical nature (adjuvants) published in recent decades and to evaluate the effectiveness of their use in the vaccination against various infections, including particularly dangerous ones. The analysis of the literature data available in PubMed, Web of Science, Scopus, eLibrary databases, indicates that the search for new substances and drugs with the ability to enhance the immune response to antigens that are part of antibacterial and antiviral registered vaccines, as well as experimental preventive drugs, is an important and promising direction. The use of various substances and compounds as adjuvants enhances the immunogenic and protective properties of vaccines, reduces the antigenic load on the human body and causes a tense immune response in individuals with reduced functioning of immune system and in the elderly. When choosing an adjuvant, it is necessary to take into account the direction of its action on the formation of both local and systemic specific immune response, depending on the nature of the pathogen.
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