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A CFSE-based assay of proliferative response of lymphocytes to stimulation with dengue viruses.
Gil, L; Alonso, R; López, C; Blanco, A; Romero, Y; Guillén, G; Hermida, L.
Afiliação
  • Gil L; Division of Vaccines, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10 600, Cuba.
Acta Virol ; 53(3): 147-52, 2009.
Article em En | MEDLINE | ID: mdl-19941395
Dengue viruses (DENVs) are human pathogens that constitute a significant threat worldwide. Since they up-regulate MHC class I molecules; the cell-mediated immunity may play an important role in the defense against viruses. In this work, we tested a CFSE-based assay in determining proliferative response of lymphocytes isolated from mice or monkeys previously immunized with various DENV antigens to in vitro stimulation with DENVs. A positive proliferative response was obtained with lymphocytes of animals immunized with either live DENV-2 or its recombinant proteins. A similar result was also obtained with CD8+ T cells from mice immunized with live DENV-1 or DENV-2 following stimulation with homologous viruses. A comparison of the carboxyfluorescein diacetate succinimidyl ester (CFSE)-based and a 3H-thymidine incorporation-based assays of proliferative response of total lymphocytes showed a fair correlation of results of both assays. However, the CSFE-based assay offers in addition the determination of contribution of the CD8+ or other subsets of T cells to total proliferative response. These results represent the first and successful application of a CFSE-based assay to the evaluation of cell-mediated immunity to DENVs. This assay might be also exploited in testing candidate DENV vaccines.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Succinimidas / Ativação Linfocitária / Vírus da Dengue / Fluoresceínas Limite: Animals Idioma: En Revista: Acta Virol Ano de publicação: 2009 Tipo de documento: Article País de afiliação: Cuba País de publicação: Eslováquia
Buscar no Google
Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Succinimidas / Ativação Linfocitária / Vírus da Dengue / Fluoresceínas Limite: Animals Idioma: En Revista: Acta Virol Ano de publicação: 2009 Tipo de documento: Article País de afiliação: Cuba País de publicação: Eslováquia