Mangada, Marlou M.Ennis, Francis A.Rothman, Alan L.2022-08-232022-08-232004-01-012017-08-22J Immunol Methods. 2004 Jan;284(1-2):89-97. doi:10.1016/j.jim.2003.10.0030022-1759 (Linking)10.1016/j.jim.2003.10.00314736419https://hdl.handle.net/20.500.14038/35055We developed an intracellular cytokine staining assay to quantify dengue specific memory T cells elicited by a primary dengue virus (DEN) infection. Peripheral blood mononuclear cells (PBMC) of volunteers who received experimental live attenuated monovalent DEN vaccines were stimulated with glutaraldehyde-inactivated dengue-infected Vero cell culture lysates from all four DEN serotypes. CD4+ T cell frequencies to previously identified MHC class II peptides were equivalent to 40-70% of the responses using virus infected cell lysates in two donors. IFN-gamma responses to DEN were detected from 0.04% to 0.45% CD4+ T cells. The highest IFN-gamma response was elicited by antigens from the homologous serotype. We detected serotype-specific and -cross reactive CD4+ T cell cytokine responses from all donors. This assay is suitable for measuring DEN specific CD4+ T cells in human PBMC from large population studies where donor haplotype is unknown or highly variant.en-USImmunityImmunology and Infectious DiseaseImmunology of Infectious DiseaseInfectious DiseaseJournal Articlehttps://escholarship.umassmed.edu/infdis_pp/26910636645infdis_pp/269