Universidad Peruana Cayetano Heredia

Standardization of a fluorescent-based quantitative adhesion assay to study attachment of Taenia solium oncosphere to epithelial cells in vitro

Mostrar el registro sencillo del ítem

dc.contributor.author Chile Andrade, Nancy
dc.contributor.author Evangelista, J.
dc.contributor.author Gilman, Robert Hugh
dc.contributor.author Arana, Y.
dc.contributor.author Palma, S.
dc.contributor.author Sterling, C.R.
dc.contributor.author García Lescano, Héctor Hugo
dc.contributor.author Gonzalez Zariquiey, Armando Emiliano
dc.contributor.author Verastegui Pimentel, Manuela Renee
dc.date.accessioned 2022-01-18T19:26:47Z
dc.date.available 2022-01-18T19:26:47Z
dc.date.issued 2012
dc.identifier.uri https://hdl.handle.net/20.500.12866/10873
dc.description.abstract To fully understand the preliminary stages of Taenia solium oncosphere attachment in the gut, adequate tools and assays are necessary to observe and quantify this event that leads to infection. A fluorescent-based quantitative adhesion assay, using biotinylated activated-oncospheres and monolayers of Chinese hamster ovary cells (CHO-K1) or human intestinal monolayer cells (INT-407, HCT-8 or HT-29), was developed to study initial events during the infection of target cells and to rapidly quantify the in vitro adhesion of T. solium oncospheres. Fluorescein streptavidin was used to identify biotinylated activated-oncospheres adhered to cells. This adherence was quantified using an automated fluorescence plate reader, and the results were expressed as fluorescence intensity values. A series of three assays were performed. The first was to identify the optimum number of biotinylated activated-oncospheres to be used in the adhesion assay. The goal of the second assay was to validate this novel method with the established oncosphere-binding system using the immunofluorescent-antibody assay (IFA) method to quantify oncosphere adhesion. A total of 10,000 biotinylated activated-oncospheres were utilized to assess the role of sera and laminin (LM) in oncosphere adherence to a CHO-K1 cell monolayer. The findings that sera and LM increase the adhesion of oncospheres to monolayer cells were similar to results that were previously obtained using the IFA method. The third assay compared the adherence of biotinylated activated-oncospheres to different types of human intestinal monolayer cells. In this case, the fluorescence intensity was greatest when using the INT-407 cell monolayer. We believe this new method of quantification offers the potential for rapid, large-scale screening to study and elucidate specific molecules and mechanisms involved in oncosphere-host cell attachment. en_US
dc.language.iso eng
dc.publisher Elsevier
dc.relation.ispartofseries Journal of Immunological Methods
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Humans en_US
dc.subject Animal Cell en_US
dc.subject In Vitro Study en_US
dc.subject Taenia Solium en_US
dc.subject Streptavidin en_US
dc.subject Immunofluorescence Test en_US
dc.subject Fluorescent Antibody Technique en_US
dc.subject Cell Adhesion en_US
dc.subject Biotinylation en_US
dc.subject Cell Adhesion en_US
dc.subject CHO Cells en_US
dc.subject Cricetinae en_US
dc.subject Cricetulus Griseus en_US
dc.subject Epithelial Cells en_US
dc.subject HT29 Cells en_US
dc.subject Intestine Epithelium Cell en_US
dc.subject Laminin en_US
dc.subject Oncosphere en_US
dc.subject Quantitative Adhesion Assay en_US
dc.subject Taenia Solium Oncosphere en_US
dc.title Standardization of a fluorescent-based quantitative adhesion assay to study attachment of Taenia solium oncosphere to epithelial cells in vitro en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.jim.2011.12.001
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.01.03
dc.relation.issn 1872-7905


Ficheros en el ítem

Ficheros Tamaño Formato Ver

No hay ficheros asociados a este ítem.

Este ítem aparece en la(s) siguiente(s) colección(ones)

Mostrar el registro sencillo del ítem

info:eu-repo/semantics/restrictedAccess Excepto si se señala otra cosa, la licencia del ítem se describe como info:eu-repo/semantics/restrictedAccess

Buscar en el Repositorio


Listar

Panel de Control

Estadísticas