A CFSE-based assay of proliferative response of lymphocytes to stimulation with dengue viruses.
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.
Gil
L
L
Division of Vaccines, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10 600, Cuba.
Alonso
R
R
López
C
C
Blanco
A
A
Romero
Y
Y
Guillén
G
G
Hermida
L
L
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002522:Chlorocebus aethiops; D003716:Dengue Virus / Q000276:immunology*; D005452:Fluoresceins; D007114:Immunization; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D013388:Succinimides; D013936:Thymidine / Q000378:metabolism
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000956:Antigens, Viral; D005452:Fluoresceins; D013388:Succinimides; D013936:Thymidine