A CFSE-based assay of proliferative response of lymphocytes to stimulation with dengue viruses

作者：Gil L^*; Alonso R; Lopez C; Blanco A; Romero Y; Guillen G; Hermida L

来源：Acta Virologica, 2009, 53(3): 147-152.

DOI：10.4149/av_2009_03_147

• cell-mediated immunity
• CFSE
• dengue viruses
• lymphocytes
• CD8 T cells

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 (3)H-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.

• 出版日期2009