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Acta Virologica Vol.53, No.3, p.147-152, 2009 |
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Title: A CFSE-based assay of proliferative response of lymphocytes to stimulation with dengue viruses | ||
Author: L. Gil, R. Alonso, C. Lopez, A. Blanco, Y. Romero, G. Guillen, L. Hermida | ||
Abstract: 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. |
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Keywords: cell-mediated immunity; CFSE, dengue viruses; lymphocytes; CD8+ T cells | ||
Year: 2009, Volume: 53, Issue: 3 | Page From: 147, Page To: 152 | |
doi:10.4149/av_2009_03_147 |
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