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The cultured Enzyme-Linked ImmunoSpot (ELISpot) assay is a functional T cell assay, which is commonly used to assess virus-specific T cell responses. The use of an in vitro expansion step before the ELISpot distinguishes such "cultured" ELISpots from "ex vivo" ELISpots. Cultured ELISpots have the advantage that lower frequency responses can be analyzed compared to ex vivo ELISpots, but do carry the associated potential distortions of the expansion phase. Cultured ELISpot assays are of value to determine silent and symptomatic transmission of the Dengue virus (DENV) in the community and to identify the correlates of a DENV-specific protective immune response. We have evaluated T cell responses to the DENV using cultured ELISpot assays with serotype-specific T cell epitopes to determine past infecting dengue virus (DENV) serotypes. The peptides used in this assay do not cross react with the Japanese encephalitis virus nor other flaviviruses. Therefore, this assay is likely to be useful in determining the past infecting DENV serotype in immune-epidemiological studies and in dengue vaccine trials.

Original publication

DOI

10.1007/978-1-4939-8567-8_14

Type

Journal article

Journal

Methods in molecular biology (Clifton, N.J.)

Publication Date

29/06/2018

Volume

1808

Pages

165 - 171

Addresses

Centre for Dengue Research, Faculty of Medical Sciences, University of Sri Jayewardenepura, Nugegoda, Sri Lanka.

Keywords

Leukocytes, Mononuclear, T-Lymphocytes, Cells, Cultured, Humans, Dengue Virus, Dengue, Peptides, Epitopes, T-Lymphocyte, Enzyme-Linked Immunospot Assay