Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos

RESUMEN La babesiosis es una de las enfermedades veterinarias más importantes transmitidas por garrapatas que afecta principalmente a animales salvajes y domésticos, y recientemente es considerada como una zoonosis emergente que se distribuye en regiones tropicales y subtropicales alrededor del mund...

Full description

Autores:
Cuy Chaparro, Laura Esperanza
Tipo de recurso:
Doctoral thesis
Fecha de publicación:
2023
Institución:
Universidad Nacional de Colombia
Repositorio:
Universidad Nacional de Colombia
Idioma:
spa
OAI Identifier:
oai:repositorio.unal.edu.co:unal/85498
Acceso en línea:
https://repositorio.unal.edu.co/handle/unal/85498
https://repositorio.unal.edu.co
Palabra clave:
630 - Agricultura y tecnologías relacionadas::636 - Producción animal
610 - Medicina y salud::616 - Enfermedades
Ticks
Erythrocyte indices
Host-parasite interactions
Immunogenicity, vaccine
Cattle-Immunology
Ácaros y garrapatas
Acari
Babesiosis
Índices de eritrocitos
Interacciones huésped-parásitos
Inmunogenicidad vacunal
Bovinos -Inmunología
Garrapatas
Babesia Bovis
Péptidos con alta capacidad de unión
Eritrocito bovino
Parasite adhesion
High activity binding peptides
Bovine erythrocyte
Rights
openAccess
License
Atribución-NoComercial 4.0 Internacional
id UNACIONAL2_f8c12ef5396e461d823dd3636524b950
oai_identifier_str oai:repositorio.unal.edu.co:unal/85498
network_acronym_str UNACIONAL2
network_name_str Universidad Nacional de Colombia
repository_id_str
dc.title.spa.fl_str_mv Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
dc.title.translated.none.fl_str_mv Identification of Babesia bovis' protein binding regions to bovine erythrocytes
title Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
spellingShingle Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
630 - Agricultura y tecnologías relacionadas::636 - Producción animal
610 - Medicina y salud::616 - Enfermedades
Ticks
Erythrocyte indices
Host-parasite interactions
Immunogenicity, vaccine
Cattle-Immunology
Ácaros y garrapatas
Acari
Babesiosis
Índices de eritrocitos
Interacciones huésped-parásitos
Inmunogenicidad vacunal
Bovinos -Inmunología
Garrapatas
Babesia Bovis
Péptidos con alta capacidad de unión
Eritrocito bovino
Parasite adhesion
High activity binding peptides
Bovine erythrocyte
title_short Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
title_full Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
title_fullStr Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
title_full_unstemmed Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
title_sort Indentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinos
dc.creator.fl_str_mv Cuy Chaparro, Laura Esperanza
dc.contributor.advisor.spa.fl_str_mv Patarroyo Gutiérrez, Manuel Alfonso
Moreno Pérez, Darwin Andrés
dc.contributor.author.spa.fl_str_mv Cuy Chaparro, Laura Esperanza
dc.contributor.orcid.spa.fl_str_mv 0000-0002-2016-2117
dc.subject.ddc.spa.fl_str_mv 630 - Agricultura y tecnologías relacionadas::636 - Producción animal
610 - Medicina y salud::616 - Enfermedades
topic 630 - Agricultura y tecnologías relacionadas::636 - Producción animal
610 - Medicina y salud::616 - Enfermedades
Ticks
Erythrocyte indices
Host-parasite interactions
Immunogenicity, vaccine
Cattle-Immunology
Ácaros y garrapatas
Acari
Babesiosis
Índices de eritrocitos
Interacciones huésped-parásitos
Inmunogenicidad vacunal
Bovinos -Inmunología
Garrapatas
Babesia Bovis
Péptidos con alta capacidad de unión
Eritrocito bovino
Parasite adhesion
High activity binding peptides
Bovine erythrocyte
dc.subject.decs.eng.fl_str_mv Ticks
Erythrocyte indices
Host-parasite interactions
Immunogenicity, vaccine
Cattle-Immunology
dc.subject.decs.spa.fl_str_mv Ácaros y garrapatas
Acari
Babesiosis
Índices de eritrocitos
Interacciones huésped-parásitos
Inmunogenicidad vacunal
Bovinos -Inmunología
dc.subject.lemb.spa.fl_str_mv Garrapatas
dc.subject.proposal.spa.fl_str_mv Babesia Bovis
Péptidos con alta capacidad de unión
Eritrocito bovino
dc.subject.proposal.eng.fl_str_mv Parasite adhesion
High activity binding peptides
Bovine erythrocyte
description RESUMEN La babesiosis es una de las enfermedades veterinarias más importantes transmitidas por garrapatas que afecta principalmente a animales salvajes y domésticos, y recientemente es considerada como una zoonosis emergente que se distribuye en regiones tropicales y subtropicales alrededor del mundo. Babesia bovis, uno de los hemoprotozoarios responsable de la babesiosis bovina, es el agente más patógeno del género Babesia que causa un impacto negativo en la industria ganadera dada las altas tasas de morbimortalidad que genera. Las principales estrategias de control y tratamiento de la babesiosis se han centrado en el uso de quimioterapéuticos dirigidos contra el parásito o de acaricidas contra el vector. Sin embargo, ante la aparición de resistencia a esta clase de productos químicos, la vacunación con organismos vivos atenuados de B. bovis se ha convertido en el principal método de control de la infección. Aunque el uso de este tipo de vacunas ha estado asociado con niveles parciales de protección, éstas tienen importantes limitaciones como una vida útil corta, riesgo de reversión de la virulencia, contaminación con otros patógenos transmitidos por sangre, falla en la inducción de inmunidad protectiva contra diferentes cepas y la pérdida de inmunogenicidad, lo que ha limitado la comercialización de la vacuna. Por lo tanto, se requieren otras estrategias alternas para mejorar las condiciones de sanidad animal bovina. El conocimiento de la biología básica del parásito en relación con la caracterización de las moléculas que éste usa para invadir a sus células diana es esencial para diseñar una medida de control eficaz contra la enfermedad, como es el caso de las vacunas. Por ende, este proyecto se encaminó a identificar proteínas de B. bovis o regiones derivadas de ellas que tengan capacidad de unirse a los eritrocitos bovinos como alternativa para contribuir al conocimiento de su biología en relación con la interacción parásito-célula. Para ello, se realizó la predicción de proteínas importantes para la invasión del parásito a sus células diana mediante la exploración in silico de los datos de transcriptoma y proteoma de B. bovis, obteniendo como resultado las proteínas BbMSA-1, BbAMA-1 y BbRON2. Posteriormente, varias regiones bajo restricción funcional y con presencia de codones seleccionados negativamente en BbMSA-1, BbAMA-1 y BbRON2 fueron inferidas a través de análisis de selección natural, con el objetivo de elegir fragmentos idóneos (regiones o péptidos de 20 residuos) y evaluar así su capacidad para unirse a los eritrocitos bovinos. Los ensayos altamente sensibles de interacción proteína-célula y péptido-célula permitieron identificar varias regiones con capacidad de unión a eritrocitos bovinos y diferentes péptidos con alta capacidad de unión (HABP, del inglés High Activity Binding Peptide) a sus células diana para BbMSA-1: 42422 (39PEGSFYDDMSKFYGAVGSFD58), 42424 (91NALIKNNPMIRPDLFNATIV110) y 42426 (150TDIVEEDREKAVEYFKKHVY169); BbAMA-1: 42437 (100YMQKFDIPRNHGSGIYVDLG119), 42438 (120GYESVGSKSYRMPVGKSPVV139) y 42443 (302SPMHPVRDAIFGKWSGGSSV321); y BbRON2: 42918 (1218SFIMVKPPALHCVLKPVETL1237). Además, los análisis de predicción de la estructura terciaria y secundaria de las moléculas permitieron evidenciar que los HABPs 42422 y 42426 de MSA-1, así como 42437 y 42438 de AMA-1, son altamente helicoidales y contienen epítopes de células B y T, mientras que los HABPs 42424 de MSA-1 y 42918 de RON2 son no estructurados estando este último, localizado en una región intrínsecamente desordenada flanqueada por dos regiones helicoidales. Este es el primer estudio que analiza y describe las regiones mínimas (definidas en este estudio por componerse de 20 aminoácidos) implicadas en la unión de las proteínas MSA-1, AMA-1 y RON2 de B. bovis a su célula diana. En estudios futuros, se explorará el potencial antigénico de dichos péptidos durante la inmunización in vivo, y se evaluará su eficacia como potenciales candidatos a vacuna. (Texto tomado de la fuente)
publishDate 2023
dc.date.issued.none.fl_str_mv 2023-12
dc.date.accessioned.none.fl_str_mv 2024-01-29T20:07:37Z
dc.date.available.none.fl_str_mv 2024-01-29T20:07:37Z
dc.type.spa.fl_str_mv Trabajo de grado - Doctorado
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/doctoralThesis
dc.type.version.spa.fl_str_mv info:eu-repo/semantics/acceptedVersion
dc.type.coar.spa.fl_str_mv http://purl.org/coar/resource_type/c_db06
dc.type.content.spa.fl_str_mv Text
dc.type.redcol.spa.fl_str_mv http://purl.org/redcol/resource_type/TD
format http://purl.org/coar/resource_type/c_db06
status_str acceptedVersion
dc.identifier.uri.none.fl_str_mv https://repositorio.unal.edu.co/handle/unal/85498
dc.identifier.instname.spa.fl_str_mv Universidad Nacional de Colombia
dc.identifier.reponame.spa.fl_str_mv Repositorio Institucional Universidad Nacional de Colombia
dc.identifier.repourl.spa.fl_str_mv https://repositorio.unal.edu.co
url https://repositorio.unal.edu.co/handle/unal/85498
https://repositorio.unal.edu.co
identifier_str_mv Universidad Nacional de Colombia
Repositorio Institucional Universidad Nacional de Colombia
dc.language.iso.spa.fl_str_mv spa
language spa
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dc.format.extent.spa.fl_str_mv [v], 36 páginas
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dc.publisher.spa.fl_str_mv Universidad Nacional de Colombia
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dc.publisher.branch.spa.fl_str_mv Universidad Nacional de Colombia - Sede Bogotá
institution Universidad Nacional de Colombia
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spelling Atribución-NoComercial 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Patarroyo Gutiérrez, Manuel Alfonso30322a85ae0ad9c1368bae791386bffeMoreno Pérez, Darwin Andrés2544f79cefd410a354e61644479f5435600Cuy Chaparro, Laura Esperanzaf92884620178c235a6a98a173bc000000000-0002-2016-21172024-01-29T20:07:37Z2024-01-29T20:07:37Z2023-12https://repositorio.unal.edu.co/handle/unal/85498Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.coRESUMEN La babesiosis es una de las enfermedades veterinarias más importantes transmitidas por garrapatas que afecta principalmente a animales salvajes y domésticos, y recientemente es considerada como una zoonosis emergente que se distribuye en regiones tropicales y subtropicales alrededor del mundo. Babesia bovis, uno de los hemoprotozoarios responsable de la babesiosis bovina, es el agente más patógeno del género Babesia que causa un impacto negativo en la industria ganadera dada las altas tasas de morbimortalidad que genera. Las principales estrategias de control y tratamiento de la babesiosis se han centrado en el uso de quimioterapéuticos dirigidos contra el parásito o de acaricidas contra el vector. Sin embargo, ante la aparición de resistencia a esta clase de productos químicos, la vacunación con organismos vivos atenuados de B. bovis se ha convertido en el principal método de control de la infección. Aunque el uso de este tipo de vacunas ha estado asociado con niveles parciales de protección, éstas tienen importantes limitaciones como una vida útil corta, riesgo de reversión de la virulencia, contaminación con otros patógenos transmitidos por sangre, falla en la inducción de inmunidad protectiva contra diferentes cepas y la pérdida de inmunogenicidad, lo que ha limitado la comercialización de la vacuna. Por lo tanto, se requieren otras estrategias alternas para mejorar las condiciones de sanidad animal bovina. El conocimiento de la biología básica del parásito en relación con la caracterización de las moléculas que éste usa para invadir a sus células diana es esencial para diseñar una medida de control eficaz contra la enfermedad, como es el caso de las vacunas. Por ende, este proyecto se encaminó a identificar proteínas de B. bovis o regiones derivadas de ellas que tengan capacidad de unirse a los eritrocitos bovinos como alternativa para contribuir al conocimiento de su biología en relación con la interacción parásito-célula. Para ello, se realizó la predicción de proteínas importantes para la invasión del parásito a sus células diana mediante la exploración in silico de los datos de transcriptoma y proteoma de B. bovis, obteniendo como resultado las proteínas BbMSA-1, BbAMA-1 y BbRON2. Posteriormente, varias regiones bajo restricción funcional y con presencia de codones seleccionados negativamente en BbMSA-1, BbAMA-1 y BbRON2 fueron inferidas a través de análisis de selección natural, con el objetivo de elegir fragmentos idóneos (regiones o péptidos de 20 residuos) y evaluar así su capacidad para unirse a los eritrocitos bovinos. Los ensayos altamente sensibles de interacción proteína-célula y péptido-célula permitieron identificar varias regiones con capacidad de unión a eritrocitos bovinos y diferentes péptidos con alta capacidad de unión (HABP, del inglés High Activity Binding Peptide) a sus células diana para BbMSA-1: 42422 (39PEGSFYDDMSKFYGAVGSFD58), 42424 (91NALIKNNPMIRPDLFNATIV110) y 42426 (150TDIVEEDREKAVEYFKKHVY169); BbAMA-1: 42437 (100YMQKFDIPRNHGSGIYVDLG119), 42438 (120GYESVGSKSYRMPVGKSPVV139) y 42443 (302SPMHPVRDAIFGKWSGGSSV321); y BbRON2: 42918 (1218SFIMVKPPALHCVLKPVETL1237). Además, los análisis de predicción de la estructura terciaria y secundaria de las moléculas permitieron evidenciar que los HABPs 42422 y 42426 de MSA-1, así como 42437 y 42438 de AMA-1, son altamente helicoidales y contienen epítopes de células B y T, mientras que los HABPs 42424 de MSA-1 y 42918 de RON2 son no estructurados estando este último, localizado en una región intrínsecamente desordenada flanqueada por dos regiones helicoidales. Este es el primer estudio que analiza y describe las regiones mínimas (definidas en este estudio por componerse de 20 aminoácidos) implicadas en la unión de las proteínas MSA-1, AMA-1 y RON2 de B. bovis a su célula diana. En estudios futuros, se explorará el potencial antigénico de dichos péptidos durante la inmunización in vivo, y se evaluará su eficacia como potenciales candidatos a vacuna. (Texto tomado de la fuente)Babesiosis is one of the most important tick-borne veterinary diseases affecting both wild and domestic animals. Recently, it has been considered also as an emerging zoonosis distributed in tropical and subtropical regions around the world. Babesia bovis, the hemoprotozoa responsible for bovine babesiosis, is the most pathogenic agent in the Babesia genus causing a negative impact on the livestock industry related to the high morbidity and mortality rates it generates. The main control and treatment strategies for babesiosis have been focused on chemotherapeutics against the parasite or acaricides against the vector. However, the emergence of resistance against these chemicals have made vaccination with live atenuated B. bovis organisms the main infection control method. Although this type of vaccines has been associated with significant levels of protection, there are limitations such as a short shelf life, risk of virulence reversal, contamination with other blood-borne pathogens, failure to induce protective immunity against different strains and immunogenicity loss. Those restrictions have limited the vaccine commercialization making alternative strategies needed to improve bovine animal welfare. Knowledge of the parasite’s basic biology and the characterization of the molecules used to invade its target cells is essential to design an effective control measure against the disease. This project was thus aimed at identifying B. bovis proteins or protein-regions with the ability to bind to bovine erythrocytes (its target cell) to elucidate the parasite-cell interaction. For this, the prediction of parasite’s important proteins for the invasion to its target cells was carried out in silico by transcriptome and proteome data exploration, which led to selecting BbMSA-1, BbAMA-1 and BbRON2 proteins. Subsequently, several regions under functional restriction and presenting negatively selected codons in BbMSA-1, BbAMA-1 and BbRON2 were inferred through natural selection analysis to choose suitable fragments (20-mer peptide regions) to evaluate their ability to bind to bovine erythrocytes. Highly sensitive protein-cell and peptide-cell interaction assays allowed the identification of several binding regions to bovine erythrocytes and different High Activity Binding Peptides (HABPs) to their target cells; for BbMSA1: peptides 42422 (39PEGSFYDDMSKFYGAVGSFD58), 42424 (91NALIKNNPMIRPDLFNATIV110 ) and 42426 (150TDIVEEDREKAVEYFKKHVY169); for BbAMA-1: peptides 42437 (100YMQKFDIPRNHGSGIYVDLG119), 42438 (120GYESVGSKSYRMPVGKSPVV139) and 42443 (302SPMHPVRDAIFGKWSGGSSV321), and for BbRON2: peptide 42918 (1218SFIMVKPPALHCVLKPVETL1237). Tertiary and secondary structure prediction analysis showed that HABPs 42422 and 42426 of MSA-1, together with 42437 and 42438 of AMA-1 are highly helical and can contain epitopes for B and T cells. MSA-1 HABP 42424 is unstructured, while RON2 HABP 42918 is in an intrinsically disordered region flanked by two helical regions. This is the first study describing the minimal regions (defined here as being 20 aa long) involved in the binding of the B. bovis MSA1, AMA-1 and RON2 proteins to their target cell. Further studies should follow to explore the antigenic potential of these peptides by in vivo immunization, to assess their efficacy as potential anti-B. bovis vaccine candidates.DoctoradoDoctora en Biotecnología[v], 36 páginasapplication/pdfspaUniversidad Nacional de ColombiaBogotá - Ciencias - Doctorado en BiotecnologíaFacultad de CienciasBogotá, ColombiaUniversidad Nacional de Colombia - Sede Bogotá630 - Agricultura y tecnologías relacionadas::636 - Producción animal610 - Medicina y salud::616 - EnfermedadesTicksErythrocyte indicesHost-parasite interactionsImmunogenicity, vaccineCattle-ImmunologyÁcaros y garrapatasAcariBabesiosisÍndices de eritrocitosInteracciones huésped-parásitosInmunogenicidad vacunalBovinos -InmunologíaGarrapatasBabesia BovisPéptidos con alta capacidad de uniónEritrocito bovinoParasite adhesionHigh activity binding peptidesBovine erythrocyteIndentificación de regiones de unión de proteínas de Babesia bovis a eritrocitos bovinosIdentification of Babesia bovis' protein binding regions to bovine erythrocytesTrabajo de grado - Doctoradoinfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_db06Texthttp://purl.org/redcol/resource_type/TDSingh B, Varikuti S, Halsey G, Volpedo G, Hamza OM, Satoskar AR. 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Sci Rep. 2017;7(1):9616.EstudiantesInvestigadoresMaestrosPúblico generalORIGINAL1057590849.2023.pdf1057590849.2023.pdfTesis de Doctorado en Biotecnologíaapplication/pdf4328501https://repositorio.unal.edu.co/bitstream/unal/85498/5/1057590849.2023.pdfd9ce33fa85d413a9268a5b1fd7cc3256MD55LICENSElicense.txtlicense.txttext/plain; charset=utf-85879https://repositorio.unal.edu.co/bitstream/unal/85498/3/license.txteb34b1cf90b7e1103fc9dfd26be24b4aMD53THUMBNAIL1057590849.2023.pdf.jpg1057590849.2023.pdf.jpgGenerated Thumbnailimage/jpeg3965https://repositorio.unal.edu.co/bitstream/unal/85498/6/1057590849.2023.pdf.jpg7193ab1f69626cb700e2d6aea0e946c3MD56unal/85498oai:repositorio.unal.edu.co:unal/854982024-08-21 23:13:00.619Repositorio Institucional Universidad Nacional de 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