Memorias do Instituto Oswaldo Cruz · 2025
Exploring BCG to deliver avidin fusion antigens from Schistosoma mansoni.
СтатьяНазвание и аннотация: Перевод в очереди
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Yamamoto Lais Sayuri, Trentini Monalisa Martins, Rodriguez Dunia, Silveira Paulo Henrique Santana, Januzzi Arthur Daniel, Carvalho Ana Carolina de Oliveira, Leite Luciana Cezar de Cerqueira, Kanno Alex Issamu
Аннотация
BACKGROUND: Bacillus Calmette-Guérin (BCG) is one of the most successful vaccines in the world and evidence suggests it can be used as a bacterial vector to deliver heterologous antigens.
OBJECTIVES: We evaluated whether BCG could be biotinylated and used as a carrier of Schistosoma mansoni antigen tetraspanin-2 (TSP-2) fused with rhizavidin, an avidin analog.
METHODS: BCG was grown and biotinylated. The recombinant protein Rzv:TSP-2 was produced and purified from Escherichia coli. The biotinylation and antigen coupling was analysed by flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot. Vaccine immunogenicity was tested in immunised mice by the assessment of lung and splenic T cells.
FINDINGS: BCG can be biotinylated, which in turn, can be coupled with Rzv:TSP-2. After a series of optimisations which involved molarity of the biotin, ratio of BCG:reagent and the concentration of Rzv:TSP-2 used, almost 50% of the bacteria were biotinylated and 35% coupled with antigen. Although a clear adjuvant effect of BCG was observed, evaluation of immune response in immunised mice demonstrated an overall low immunogenicity of the BCG-Rzv:TSP-2.
MAIN CONCLUSION: These results demonstrated the use of BCG as a carrier of avidin-tagged antigens. Further optimisations are needed in order to strengthen the stability of tagged proteins in order to produce antigen-specific immune responses.
OBJECTIVES: We evaluated whether BCG could be biotinylated and used as a carrier of Schistosoma mansoni antigen tetraspanin-2 (TSP-2) fused with rhizavidin, an avidin analog.
METHODS: BCG was grown and biotinylated. The recombinant protein Rzv:TSP-2 was produced and purified from Escherichia coli. The biotinylation and antigen coupling was analysed by flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot. Vaccine immunogenicity was tested in immunised mice by the assessment of lung and splenic T cells.
FINDINGS: BCG can be biotinylated, which in turn, can be coupled with Rzv:TSP-2. After a series of optimisations which involved molarity of the biotin, ratio of BCG:reagent and the concentration of Rzv:TSP-2 used, almost 50% of the bacteria were biotinylated and 35% coupled with antigen. Although a clear adjuvant effect of BCG was observed, evaluation of immune response in immunised mice demonstrated an overall low immunogenicity of the BCG-Rzv:TSP-2.
MAIN CONCLUSION: These results demonstrated the use of BCG as a carrier of avidin-tagged antigens. Further optimisations are needed in order to strengthen the stability of tagged proteins in order to produce antigen-specific immune responses.