Ex Vivo ELISpot Assay to Investigate Dengue Virus Specific T-Cell Responses

Enzyme-Linked ImmunoSpot (ELISpot) assay is a functional T cell assay which is commonly used to assess virus-specific T cell responses. We have used ex vivo ELISpot responses to investigate the role of dengue virus (DENV) specific T cells in acute dengue infection, to evaluate their role in possible disease pathogenesis and protection. We have also used ex vivo IFNγ ELISpot assays to determine the functionality of T cell responses in those who were previously naturally infected with dengue and we have determined the frequency of DENV-specific memory T cells responses in relation to past clinical disease severity. We have also evaluated ELISpot supernatants to study multiple cytokines produced by DENV-specific T cells, in instances where there are fewer numbers of cells available for multiple assays. ELISpot assays are likely to be helpful in large-scale dengue vaccine trials to assess the immunogenicity of the vaccines.