Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da UFMG |
| Texto Completo: | https://doi.org/10.1128/mbio.03478-21 http://hdl.handle.net/1843/62488 https://orcid.org/0000-0002-2588-2842 https://orcid.org/0000-0002-9493-1740 |
Resumo: | CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico |
| id |
UFMG_0438a1fddc54b62d78f1108d7b9d3329 |
|---|---|
| oai_identifier_str |
oai:repositorio.ufmg.br:1843/62488 |
| network_acronym_str |
UFMG |
| network_name_str |
Repositório Institucional da UFMG |
| repository_id_str |
|
| spelling |
2024-01-08T20:00:06Z2024-01-08T20:00:06Z2022-01-25131e03478-2117https://doi.org/10.1128/mbio.03478-212150-7511http://hdl.handle.net/1843/62488https://orcid.org/0000-0002-2588-2842https://orcid.org/0000-0002-9493-1740CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisTrans-sialidases (TS) are unusual enzymes present on the surface of Trypanosoma cruzi, the causative agent of Chagas disease. Encoded by the largest gene family in the T. cruzi genome, only few members of the TS family have catalytic activity. Active trans-sialidases (aTS) are responsible for transferring sialic acid from host glycoconjugates to mucins, also present on the parasite surface. The existence of several copies of TS genes has impaired the use of reverse genetics to study this highly polymorphic gene family. Using CRISPR-Cas9, we generated aTS knockout cell lines displaying undetectable levels of TS activity, as shown by sialylation assays and labeling with antibodies that recognize sialic acid-containing mucins. In vitro infection assays showed that disruption of aTS genes does not affect the parasite’s capacity to invade cells or to escape from the parasitophorous vacuole but resulted in impaired differentiation of amastigotes into trypomastigotes and parasite egress from the cell. When inoculated into mice, aTS mutants were unable to establish infection even in the highly susceptible gamma interferon (IFN-γ) knockout mice. Mice immunized with aTS mutants were fully protected against a challenge infection with the virulent T. cruzi Y strain. Altogether, our results confirmed the role of aTS as a T. cruzi virulence factor and indicated that aTS play a major role during the late stages of intracellular development and parasite egress. Notably, mutants lacking TS activity are completely avirulent in animal models of infection and may be used as a live attenuated vaccine against Chagas disease.engUniversidade Federal de Minas GeraisUFMGBrasilICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIAICB - DEPARTAMENTO DE PARASITOLOGIAICB - INSTITUTO DE CIÊNCIAS BIOLOGICASMicrobial GeneticsTrypanosoma cruziVacinasVirulênciaTrypanosoma cruziTrans-sialidaseVirulenceVaccineCRISPR-Cas9Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasitesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttps://journals.asm.org/doi/10.1128/mbio.03478-21Gabriela de A. Burle-CaldasJúlia T. de CastroNailma Aprigio Dos SantosViviane Grazielle da SilvaFernanda L. B. MuggeSergio SchenkmanRicardo T. GazzinelliSantuza Maria Ribeiro TeixeiraAntônio Edson R. OliveiraMilton C. A. PereiraJoão Luís Reis-CunhaAnderson Coqueiro dos SantosDawidson Assis GomesDaniella C. BartholomeuNilmar S. Morettiapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSELicense.txtLicense.txttext/plain; charset=utf-82042https://repositorio.ufmg.br/bitstream/1843/62488/1/License.txtfa505098d172de0bc8864fc1287ffe22MD51ORIGINALDisruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites.pdfDisruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites.pdfapplication/pdf3278368https://repositorio.ufmg.br/bitstream/1843/62488/2/Disruption%20of%20Active%20Trans-Sialidase%20Genes%20Impairs%20Egress%20from%20Mammalian%20Host%20Cells%20and%20Generates%20Highly%20Attenuated%20Trypanosoma%20cruzi%20Parasites.pdf86b72dc1d9f92422d4eb4242670460e9MD521843/624882024-01-08 17:00:07.295oai:repositorio.ufmg.br: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Repositório de PublicaçõesPUBhttps://repositorio.ufmg.br/oaiopendoar:2024-01-08T20:00:07Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false |
| dc.title.pt_BR.fl_str_mv |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| title |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| spellingShingle |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites Gabriela de A. Burle-Caldas Trypanosoma cruzi Trans-sialidase Virulence Vaccine CRISPR-Cas9 Trypanosoma cruzi Vacinas Virulência |
| title_short |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| title_full |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| title_fullStr |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| title_full_unstemmed |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| title_sort |
Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites |
| author |
Gabriela de A. Burle-Caldas |
| author_facet |
Gabriela de A. Burle-Caldas Júlia T. de Castro Nailma Aprigio Dos Santos Viviane Grazielle da Silva Fernanda L. B. Mugge Sergio Schenkman Ricardo T. Gazzinelli Santuza Maria Ribeiro Teixeira Antônio Edson R. Oliveira Milton C. A. Pereira João Luís Reis-Cunha Anderson Coqueiro dos Santos Dawidson Assis Gomes Daniella C. Bartholomeu Nilmar S. Moretti |
| author_role |
author |
| author2 |
Júlia T. de Castro Nailma Aprigio Dos Santos Viviane Grazielle da Silva Fernanda L. B. Mugge Sergio Schenkman Ricardo T. Gazzinelli Santuza Maria Ribeiro Teixeira Antônio Edson R. Oliveira Milton C. A. Pereira João Luís Reis-Cunha Anderson Coqueiro dos Santos Dawidson Assis Gomes Daniella C. Bartholomeu Nilmar S. Moretti |
| author2_role |
author author author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Gabriela de A. Burle-Caldas Júlia T. de Castro Nailma Aprigio Dos Santos Viviane Grazielle da Silva Fernanda L. B. Mugge Sergio Schenkman Ricardo T. Gazzinelli Santuza Maria Ribeiro Teixeira Antônio Edson R. Oliveira Milton C. A. Pereira João Luís Reis-Cunha Anderson Coqueiro dos Santos Dawidson Assis Gomes Daniella C. Bartholomeu Nilmar S. Moretti |
| dc.subject.por.fl_str_mv |
Trypanosoma cruzi Trans-sialidase Virulence Vaccine CRISPR-Cas9 |
| topic |
Trypanosoma cruzi Trans-sialidase Virulence Vaccine CRISPR-Cas9 Trypanosoma cruzi Vacinas Virulência |
| dc.subject.other.pt_BR.fl_str_mv |
Trypanosoma cruzi Vacinas Virulência |
| description |
CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico |
| publishDate |
2022 |
| dc.date.issued.fl_str_mv |
2022-01-25 |
| dc.date.accessioned.fl_str_mv |
2024-01-08T20:00:06Z |
| dc.date.available.fl_str_mv |
2024-01-08T20:00:06Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1843/62488 |
| dc.identifier.doi.pt_BR.fl_str_mv |
https://doi.org/10.1128/mbio.03478-21 |
| dc.identifier.issn.pt_BR.fl_str_mv |
2150-7511 |
| dc.identifier.orcid.pt_BR.fl_str_mv |
https://orcid.org/0000-0002-2588-2842 https://orcid.org/0000-0002-9493-1740 |
| url |
https://doi.org/10.1128/mbio.03478-21 http://hdl.handle.net/1843/62488 https://orcid.org/0000-0002-2588-2842 https://orcid.org/0000-0002-9493-1740 |
| identifier_str_mv |
2150-7511 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartof.pt_BR.fl_str_mv |
Microbial Genetics |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
| dc.publisher.initials.fl_str_mv |
UFMG |
| dc.publisher.country.fl_str_mv |
Brasil |
| dc.publisher.department.fl_str_mv |
ICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIA ICB - DEPARTAMENTO DE PARASITOLOGIA ICB - INSTITUTO DE CIÊNCIAS BIOLOGICAS |
| publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
| dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFMG instname:Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
| instname_str |
Universidade Federal de Minas Gerais (UFMG) |
| instacron_str |
UFMG |
| institution |
UFMG |
| reponame_str |
Repositório Institucional da UFMG |
| collection |
Repositório Institucional da UFMG |
| bitstream.url.fl_str_mv |
https://repositorio.ufmg.br/bitstream/1843/62488/1/License.txt https://repositorio.ufmg.br/bitstream/1843/62488/2/Disruption%20of%20Active%20Trans-Sialidase%20Genes%20Impairs%20Egress%20from%20Mammalian%20Host%20Cells%20and%20Generates%20Highly%20Attenuated%20Trypanosoma%20cruzi%20Parasites.pdf |
| bitstream.checksum.fl_str_mv |
fa505098d172de0bc8864fc1287ffe22 86b72dc1d9f92422d4eb4242670460e9 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 |
| repository.name.fl_str_mv |
Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG) |
| repository.mail.fl_str_mv |
|
| _version_ |
1803589147697348608 |