Obtaining from the cell walls of Saccharomyces cerevisiae yeast and assessing their adjuvant properties in a subunit vaccine model

Abstract

Background. Polysaccharides are known to possess adjuvant properties, they are biodegradable, safe, and are of low-labor production. In this regard, the development of polysaccharide-based adjuvants is an urgent task.The aim. To develop a method for obtaining mannans from the cell walls of Saccharomyces cerevisiae yeast and to study their adjuvant properties using subunit vaccine model.Materials and methods. The preparation of mannans was obtained from the Saccharomyces cerevisiae yeast by enzymatic and alkaline hydrolysis. Its adjuvant properties were assessed in BALB/c mice immunized with the recombinant receptor-binding domain (RBD) of the SARS-CoV-2 (S) protein (Delta (B.1.617.2)). The titers of specific antibodies in the blood sera were determined by ELISA assays using the recombinant RBD (Wuhan-Hu-1 and Delta), and the recombinant (S) protein (Wuhan-Hu-1, Delta and Omicron) as antigens. The titers of virus-neutralizing antibodies were determined using virus-neutralization tests with the SARS-CoV-2 virus strains Wuhan – hCoV19/Australia/VIC01/2020 (Wuhan-Hu-1), Delta – hCoV-19/Russia/PSK-2804/2021 (Delta (B.1.617.2)), and Omicron 1 – hCoV-19/Russia/Moscow171619-031221/2021 (Omicron (B.1.1.529)).Results. The developed scheme allowed for obtaining up to 200 mg of mannans from 10 g of yeast cell debris. Double, with a two-week interval, immunization with RBD (50 μg) in combination with mannans (40 μg and 10 μg) induced the production of specific antibodies in titers from 1:2477330 to 1:188360. The titer of virus-neutralizing antibodies to the Delta – hCoV-19/Russia/PSK-2804/2021 was 1:485 (40 μg of mannans per mouse).Conclusions. We developed a scheme for obtaining a low-toxic preparation of mannans from the Saccharomyces cerevisiae yeast. The highest adjuvant activity was achieved when using mannans at the dose of 40 µg per mouse. Blood sera obtained from the immunized animals neutralized both homologous and heterologous SARS-CoV-2 strains.