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dc.contributor.advisorDíaz Arévalo, Diana 
dc.creatorCubides Amézquita, Jenner Rodrigo 
dc.date.accessioned2019-08-21T20:44:18Z
dc.date.available2019-08-21T20:44:18Z
dc.date.created2019-06-05
dc.date.issued2019
dc.identifier.urihttp://repository.urosario.edu.co/handle/10336/20148
dc.descriptionA pesar de que varios candidatos a la vacuna contra la malaria se encuentran actualmente ensayos de fase clínica 2 y 3, sus mecanismos de protección inmunológica no se comprenden completamente y la duración de la protección de la vacuna se desconoce en gran medida. El modelo del mono Aotus spp. ha sido recomendado por la Organización Mundial de la Salud para el estudio de los candidatos a la vacuna contra la malaria debido a las fuertes similitudes con la patología y los antecedentes genéticos observados en humanos. Sin embargo, la falta de anticuerpos específicos para los marcadores de superficie molecular de las células inmunes en Aotus, ha retrasado los avances en la investigación de la malaria. Entre los anticuerpos monoclonales comerciales (mAbs) para los marcadores moleculares de células T de memoria CD19 +, CD27 + B y CD4 +, CD45RO +, solo los del clon SK3 reconocieron las células T Aotus CD4. En este trabajo, los enfoques bioinformáticos fueron usados para diseñar péptidos antigénicos que corresponden a las regiones extracelulares de las proteínas de membrana CD19, CD27, CD4 y CD45RO, para producir mAbs. 1746 clones de hibridoma resultantes reconocieron los marcadores de superficie molecular de las células inmunes por citometría de flujo y el 30% de ellos se unen al péptido sintético por ELISA. Los mAbs, CD194G12A3G3, CD275F11C11, CD45H3D10 y CD45RO3A8G1 se unieron al 17,7%, 40,1%, 27,4 y 51% de los PBMC de Aotus con alta afinidad (100 ng / 106 células) pero solo mostraron afinidad media a las células humanas (300 ng / 106 células) en análisis FACS. En los ensayos de doble marcaje de las células B y T, mostró que los mAbs CD194G12A3G3 y CD275F11C11 reconocieron el 15,9%, y CD45H3D10 y CD45RO3A8G1 se unieron al 20,6% de las PBMC de Aotus, lo que sugiere que los mAbs reconocieron los marcadores de proteínas de membrana de las células B y T. Estos mAbs son útiles para la identificación y el seguimiento de las células de memoria en el modelo de Aotus para dilucidar qué células inmunes humanas pueden mediar la protección contra la malaria.
dc.description.abstractEven though several malaria vaccine candidates are currently in clinical phase 2, and 3 trials, their mechanisms of immune protection are not fully understood, and durations of vaccine protection are largely unknown. The Aotus monkey model has been recommended by the World Health Organization for the study of malaria vaccine candidates because of strong similarities to pathology and genetic background observed in humans. However, the lack of antibodies specific for molecular surface markers of immune cells in Aotus have delayed advances in malaria research. Among commercial monoclonal antibodies (mAbs) for human CD19+, CD27+ B cell and CD4+, CD45RO+ memory T cell molecular markers, only those of the clone SK3 recognized Aotus CD4 T cells. Here, bioinformatics approaches were used to design antigenic peptides that correspond to the extracellular regions of the membrane proteins CD19, CD27, CD4, and CD45RO, to produce mAbs. 1746 resulting hybridoma clones recognized molecular surface markers of immune cells by flow cytometry and 30% of them bond to the synthetic peptide by ELISA. The mAbs, CD194G12A3G3, CD275F11C11, CD45H3D10, and CD45RO3A8G1 bound to 17.7%, 40.1%, 27.4 and 51% of the Aotus’ PBMCs with high affinity (100 ng/106 cells) but displayed only medium affinity to human cells (300 ng/106 cells) in FACS analyses. Double staining of B and T cells showed that the mAbs CD194G12A3G3 and CD275F11C11 recognized 15.9%, and CD45H3D10 and CD45RO3A8G1 bound to 20.6% of Aotus’ PBMCs, suggesting that the mAbs recognized membrane protein markers of memory B and T cells. These mAbs are useful for the identification and tracking of memory cells in the Aotus model to elucidate which human immune cells may mediate protection against malaria
dc.format.mimetypeapplication/pdf
dc.language.isospa
dc.rightsAtribución-NoComercial-SinDerivadas 2.5 Colombia
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/2.5/co/
dc.sourceinstname:Universidad del Rosario
dc.sourcereponame:Repositorio Institucional EdocUR
dc.subjectInmunología
dc.subjectAnticuerpo Monoclonal
dc.subjectCitometría de flujo
dc.subject.ddcFisiología humana 
dc.subject.lembInmunología
dc.subject.lembAnticuerpos
dc.subject.lembCitometría de flujo
dc.titleProducción de anticuerpos monoclonales que reconozcan proteínas de membrana en células de memoria en aotus spp
dc.typemasterThesis
dc.publisherUniversidad del Rosario
dc.creator.degreeMagíster en Ciencias con Énfasis en Genética Humana
dc.publisher.programMaestría en Ciencias con Énfasis en Genética Humana
dc.publisher.departmentEscuela de Medicina y Ciencias de la Salud
dc.title.alternativeMonoclonal Antibodies for the Tracking of Aotus spp. Memory T and B Cells
dc.rights.accesRightsinfo:eu-repo/semantics/embargoedAccess
dc.type.spaTesis de maestría
dc.rights.accesoRestringido (Temporalmente bloqueado)
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dc.rights.licenciaEL AUTOR, manifiesta que la obra objeto de la presente autorización es original y la realizó sin violar o usurpar derechos de autor de terceros, por lo tanto la obra es de exclusiva autoría y tiene la titularidad sobre la misma. PARGRAFO: En caso de presentarse cualquier reclamación o acción por parte de un tercero en cuanto a los derechos de autor sobre la obra en cuestión, EL AUTOR, asumirá toda la responsabilidad, y saldrá en defensa de los derechos aquí autorizados; para todos los efectos la universidad actúa como un tercero de buena fe. EL AUTOR, autoriza a LA UNIVERSIDAD DEL ROSARIO, para que en los términos establecidos en la Ley 23 de 1982, Ley 44 de 1993, Decisión andina 351 de 1993, Decreto 460 de 1995 y demás normas generales sobre la materia, utilice y use la obra objeto de la presente autorización. -------------------------------------- POLITICA DE TRATAMIENTO DE DATOS PERSONALES. Declaro que autorizo previa y de forma informada el tratamiento de mis datos personales por parte de LA UNIVERSIDAD DEL ROSARIO para fines académicos y en aplicación de convenios con terceros o servicios conexos con actividades propias de la academia, con estricto cumplimiento de los principios de ley. Para el correcto ejercicio de mi derecho de habeas data cuento con la cuenta de correo habeasdata@urosario.edu.co, donde previa identificación podré solicitar la consulta, corrección y supresión de mis datos.
dc.type.documentTesis
dc.creator.degreetypeFull time
dc.contributor.coadvisorPatarroyo, Manuel A. 


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Atribución-NoComercial-SinDerivadas 2.5 Colombia
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