Molecular and biological characterization of an Asian-American isolate of chikungunya virus

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dc.contributor.advisorCastellanos Parra, Jaime Eduardo
dc.contributor.advisorCalvo Tapiero, Eliana Patricia
dc.contributor.authorArchila Hernández, Edwin Darío
dc.contributor.orcidArchila Hernández, Edwin Darío [0000-0002-8633-8031]
dc.date.accessioned2022-03-25T14:46:15Z
dc.date.available2022-03-25T14:46:15Z
dc.date.issued2021
dc.description.abstractBackground: Chikungunya virus is an arthropod-transmitted virus that causes chikungunya fever, a disease characterized by severe muscle and joint pain. The virus was introduced to the Americas in 2013 and caused approximately 2.7 million cases during the subsequent two years. The lack of knowledge regarding the biological behavior of the viral strains circulating during the outbreak has motivated the characterization of a virus isolated in Colombia in 2015. Methods: The full genomes of isolated and cultured viruses were obtained using next-generation sequencing. The infective and replicative capacities and their impact on viability were evaluated in three different cell lines, HEK293T, Huh-7, and MRC-5, to describe the biological behavior. The infection percentages were determined by flow cytometry, and cytopathic effects were evaluated by conducting a resazurin fluorescent metabolic assay. The viral yield of each cell line was quantified by conducting a virus plaque formation assay. The virus proteins and RNA kinetics were then evaluated by western blotting and Reverse transcription -qPCR. Results: The COL7624 isolate was clustered with other American and Caribbean sequences in the Asian American lineage. The T669A substitution in E2 distinguished it from other Colombian sequences reported in 2014. After 48 hpi, the three cell lines analyzed reached infection percentages exceeding 65%, generating a high load of infectious viral progeny. The infection kinetics indicated that the replication peak occurred around 24 hpi, although gRNA could be detected in the supernatant from 4 hpi onwards. Infection caused the overexpression of interferon and proinflammatory cytokines, such as IL-1, TNF-α, and IL-8. Conclusions: The COL7624 isolate exhibited a high infective and replicative capacity and induced the activation of the cellular immune response, similar to isolates belonging to the other genotypes.spa
dc.description.abstractenglishBackground: Chikungunya virus is an arthropod-transmitted virus that causes chikungunya fever, a disease characterized by severe muscle and joint pain. The virus was introduced to the Americas in 2013 and caused approximately 2.7 million cases during the subsequent two years. The lack of biological knowledge regarding the behavior of the viral strains circulating during the outbreak has motivated the characterization of a virus isolated in Colombia in 2015. Methods: The full genomes of isolated and cultured viruses were obtained using next-generation sequencing. The infective and replicative capacities and their impact on viability were evaluated in three different cell lines, HEK293T, Huh-7, and MRC-5, to describe the biological behavior. The infection percentages were determined by flow cytometry, and cytopathic effects were evaluated by conducting a resazurin fluorescent metabolic assay. The viral yield of each cell line was quantified by conducting a virus plaque formation assay. The virus proteins and RNA kinetics were then evaluated by western blotting and Reverse transcription-qPCR. Results: The COL7624 isolate was clustered with other American and Caribbean sequences in the Asian American lineage. The T669A substitution in E2 distinguished it from other Colombian sequences reported in 2014. After 48 hpi, the three cell lines analyzed reached infection percentages exceeding 65%, generating a high load of infectious viral progeny. The infection kinetics indicated that the replication peak occurred around 24 hpi, although gRNA could be detected in the supernatant from 4 hpi onwards. Infection caused the overexpression of interferon and proinflammatory cytokines, such as IL-1, TNF-α, and IL-8. Conclusions: The COL7624 isolate exhibited a high infective and replicative capacity and induced the activation of the cellular immune response, similar to isolates belonging to the other genotypes.eng
dc.description.degreelevelMaestríaspa
dc.description.degreenameMagíster en Ciencias Básicas Biomédicasspa
dc.format.mimetypeapplication/pdf
dc.identifier.instnameinstname:Universidad El Bosquespa
dc.identifier.reponamereponame:Repositorio Institucional Universidad El Bosquespa
dc.identifier.repourlrepourl:https://repositorio.unbosque.edu.co
dc.identifier.urihttps://hdl.handle.net/20.500.12495/7363
dc.language.isoeng
dc.publisher.facultyFacultad de Medicinaspa
dc.publisher.grantorUniversidad El Bosquespa
dc.publisher.programMaestría en Ciencias Básicas Biomédicasspa
dc.rightsAtribución-NoComercial-CompartirIgual 4.0 Internacional*
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.rights.accessrightshttps://purl.org/coar/access_right/c_abf2
dc.rights.localAcceso abiertospa
dc.rights.urihttps://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectGenomaspa
dc.subjectVirus chikungunyaspa
dc.subjectReplicaciónspa
dc.subjectInfecciónspa
dc.subject.keywordsInfectionspa
dc.subject.keywordsReplicationspa
dc.subject.keywordsChikungunya virusspa
dc.subject.keywordsGenomespa
dc.subject.nlmW 50
dc.titleMolecular and biological characterization of an Asian-American isolate of chikungunya virusspa
dc.title.translatedMolecular and biological characterization of an Asian-American isolate of chikungunya virusspa
dc.type.coarhttps://purl.org/coar/resource_type/c_bdcc
dc.type.coarversionhttps://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.driverinfo:eu-repo/semantics/masterThesis
dc.type.hasversioninfo:eu-repo/semantics/acceptedVersion
dc.type.localTesis/Trabajo de grado - Monografía - Maestríaspa

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Maestría en Ciencias Básicas Biomédicas