Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis
Surface proteins bound to the cell membrane by glycosylphosphatidylinositol (GPI) anchors are considered essential for the survival of pathogenic protozoans. In the case of the tick-transmitted hemoparasite Babesia bovis, the most virulent causative agent of bovine babesiosis, the GPI-anchored prote...
| Autores principales: | , , , , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2021
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/8610 https://www.sciencedirect.com/science/article/abs/pii/S0304401720302557 https://doi.org/10.1016/j.vetpar.2020.109275 |
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| author | Flores, Daniela Rodriguez, Anabel Elisa Tomazic, Mariela Luján Torioni, Susana Marta Echaide, Ignacio Eduardo Zamorano, Patricia Ines Langellotti, Cecilia Ana Araujo, Flabio Ribeiro de Rolls, Peter Schnittger, Leonhard Florin-Christensen, Mónica |
| author_browse | Araujo, Flabio Ribeiro de Echaide, Ignacio Eduardo Flores, Daniela Florin-Christensen, Mónica Langellotti, Cecilia Ana Rodriguez, Anabel Elisa Rolls, Peter Schnittger, Leonhard Tomazic, Mariela Luján Torioni, Susana Marta Zamorano, Patricia Ines |
| author_facet | Flores, Daniela Rodriguez, Anabel Elisa Tomazic, Mariela Luján Torioni, Susana Marta Echaide, Ignacio Eduardo Zamorano, Patricia Ines Langellotti, Cecilia Ana Araujo, Flabio Ribeiro de Rolls, Peter Schnittger, Leonhard Florin-Christensen, Mónica |
| author_sort | Flores, Daniela |
| collection | INTA Digital |
| description | Surface proteins bound to the cell membrane by glycosylphosphatidylinositol (GPI) anchors are considered essential for the survival of pathogenic protozoans. In the case of the tick-transmitted hemoparasite Babesia bovis, the most virulent causative agent of bovine babesiosis, the GPI-anchored proteome was recently unraveled by an in silico approach. In this work, one of the identified proteins, GASA-1 (GPI-Anchored Surface Antigen-1), was thoroughly characterized. GASA-1 is 179 aa long and has the characteristic features of a GPI-anchored protein, including a signal peptide, a hydrophilic core and a hydrophobic tail that harbors a GPI anchor signal. Transcriptomic analysis shows that it is expressed in pathogenic and attenuated B. bovis strains. Notably, the gasa-1 gene has syntenic counterparts in B. bigemina and B. ovata, which also encode GPI-anchored proteins. This is highly unusual since all piroplasmid GPI-anchored proteins described so far have been found to be species-specific. Sequencing of gasa-1 alleles from B. bovis geographical isolates originating from Argentina, USA, Brazil, Mexico and Australia showed over 98 % identity in both nucleotide and amino acid sequences. A recombinant form of GASA-1 (rGASA-1) was generated in E. coli and anti-rGASA-1 antibodies were raised in mice. Fixed and live immunofluorescence assays showed that GASA-1 is expressed in in vitro cultured B. bovis merozoites and surface-exposed. Moreover, incubation of B. bovis in vitro cultures with anti-GASA-1 antibodies partially, but significantly, reduced erythrocyte invasion, indicating that this protein bears neutralization-sensitive antibody epitopes. Splenocytes of rGASA-1-inoculated mice showed a specific proliferative response when exposed to the recombinant protein, indicating that GASA-1 bears T-cell epitopes. Finally, sera from a group of B. bovis-infected cattle reacted with the recombinant protein, demonstrating that GASA-1 is expressed during natural infection of bovines with B. bovis, and suggesting that it is immunodominant. The high degree of conservation among B. bovis isolates and the presence of syntenic genes in other Babesia species suggest a relevant role of GASA-1 and GASA-1-like proteins for parasite survival, especially considering that, due to their surface location, they are exposed to the selection pressure of the host immune system. The highlighted features of GASA-1 make it an interesting candidate for the development of vaccines against bovine babesiosis. |
| format | Artículo |
| id | INTA8610 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA86102021-01-18T15:32:04Z Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis Flores, Daniela Rodriguez, Anabel Elisa Tomazic, Mariela Luján Torioni, Susana Marta Echaide, Ignacio Eduardo Zamorano, Patricia Ines Langellotti, Cecilia Ana Araujo, Flabio Ribeiro de Rolls, Peter Schnittger, Leonhard Florin-Christensen, Mónica Babesiosis Vacuna Antígenos Glicosiltransferasas Babesia Bovis Vaccines Antigens Glycosyltransferases Surface proteins bound to the cell membrane by glycosylphosphatidylinositol (GPI) anchors are considered essential for the survival of pathogenic protozoans. In the case of the tick-transmitted hemoparasite Babesia bovis, the most virulent causative agent of bovine babesiosis, the GPI-anchored proteome was recently unraveled by an in silico approach. In this work, one of the identified proteins, GASA-1 (GPI-Anchored Surface Antigen-1), was thoroughly characterized. GASA-1 is 179 aa long and has the characteristic features of a GPI-anchored protein, including a signal peptide, a hydrophilic core and a hydrophobic tail that harbors a GPI anchor signal. Transcriptomic analysis shows that it is expressed in pathogenic and attenuated B. bovis strains. Notably, the gasa-1 gene has syntenic counterparts in B. bigemina and B. ovata, which also encode GPI-anchored proteins. This is highly unusual since all piroplasmid GPI-anchored proteins described so far have been found to be species-specific. Sequencing of gasa-1 alleles from B. bovis geographical isolates originating from Argentina, USA, Brazil, Mexico and Australia showed over 98 % identity in both nucleotide and amino acid sequences. A recombinant form of GASA-1 (rGASA-1) was generated in E. coli and anti-rGASA-1 antibodies were raised in mice. Fixed and live immunofluorescence assays showed that GASA-1 is expressed in in vitro cultured B. bovis merozoites and surface-exposed. Moreover, incubation of B. bovis in vitro cultures with anti-GASA-1 antibodies partially, but significantly, reduced erythrocyte invasion, indicating that this protein bears neutralization-sensitive antibody epitopes. Splenocytes of rGASA-1-inoculated mice showed a specific proliferative response when exposed to the recombinant protein, indicating that GASA-1 bears T-cell epitopes. Finally, sera from a group of B. bovis-infected cattle reacted with the recombinant protein, demonstrating that GASA-1 is expressed during natural infection of bovines with B. bovis, and suggesting that it is immunodominant. The high degree of conservation among B. bovis isolates and the presence of syntenic genes in other Babesia species suggest a relevant role of GASA-1 and GASA-1-like proteins for parasite survival, especially considering that, due to their surface location, they are exposed to the selection pressure of the host immune system. The highlighted features of GASA-1 make it an interesting candidate for the development of vaccines against bovine babesiosis. Instituto de Patobiología Fil: Flores, Daniela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Rodriguez, Anabel Elisa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Tomazic, Mariela Luján. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Torioni, Susana Marta. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Echaide, Ignacio Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Zamorano, Patricia Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Langellotti, Cecilia Ana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Araujo, Flabio Ribeiro de. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Corte; Brasil Fil: Rolls, Peter. Tick Fever Centre. Department of Agriculture & Fisheries; Australia Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Flores, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Tomazic, Mariela Luján. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina Fil: Florin-Christensen, Monica. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina 2021-01-18T15:19:34Z 2021-01-18T15:19:34Z 2020-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/8610 https://www.sciencedirect.com/science/article/abs/pii/S0304401720302557 0304-4017 https://doi.org/10.1016/j.vetpar.2020.109275 eng info:eu-repograntAgreement/INTA/2019-PD-E5-I102-001/2019-PD-E5-I102-001/AR./Desarrollo de vacunas y tecnologías para mejorar las estrategias profilácticas y terapéuticas de las enfermedades que afectan la producción animal y la salud pública info:eu-repograntAgreement/INTA/2019-PD-E5-I105-001/2019-PD-E5-I105-001/AR./Patógenos animales: su interacción con el hospedador y el medio ambiente. Impacto en productividad, ecosistemas, sanidad animal y salud pública en el marco “Una Salud” info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Veterinary parasitology 287 : 109275. (Noviembre 2020) |
| spellingShingle | Babesiosis Vacuna Antígenos Glicosiltransferasas Babesia Bovis Vaccines Antigens Glycosyltransferases Flores, Daniela Rodriguez, Anabel Elisa Tomazic, Mariela Luján Torioni, Susana Marta Echaide, Ignacio Eduardo Zamorano, Patricia Ines Langellotti, Cecilia Ana Araujo, Flabio Ribeiro de Rolls, Peter Schnittger, Leonhard Florin-Christensen, Mónica Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title | Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title_full | Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title_fullStr | Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title_full_unstemmed | Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title_short | Characterization of GASA-1, a new vaccine candidate antigen of Babesia bovis |
| title_sort | characterization of gasa 1 a new vaccine candidate antigen of babesia bovis |
| topic | Babesiosis Vacuna Antígenos Glicosiltransferasas Babesia Bovis Vaccines Antigens Glycosyltransferases |
| url | http://hdl.handle.net/20.500.12123/8610 https://www.sciencedirect.com/science/article/abs/pii/S0304401720302557 https://doi.org/10.1016/j.vetpar.2020.109275 |
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