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Ex Vivo ELISpot Assay to Investigate iNKT Cell Responses in Acute Dengue Infection

Achala Indika Kamaladasa1, Gathsaurie Neelika Malavige2,3

  • 1Centre for Dengue Research, Faculty of Medical Sciences, University of Sri Jayewardenepura, Nugegoda, Sri Lanka.

Methods in Molecular Biology (Clifton, N.J.)
|June 30, 2018
PubMed

T cell receptors of the invariant Natural Killer T (iNKT) cells are downregulated with antigen stimulation. Therefore, identification of this cell population with flow cytometry in functionality studies is challenging. iNKT cells are known to produce both Th1 and Th2 cytokines immediately upon antigen stimulation. Therefore, we have used an ELISpot assay to determine the production of IFN-ɣ and IL-4 after stimulation with KRN7000, which has shown to bind to CD1d molecules and activate iNKTs in a similar fashion as α-GalCer. In this study, we observed that peripheral iNKT cells in patients with acute dengue show distinct production of IFN-ɣ, but not IL-4 with KRN7000 stimulation.

Keywords:
IFN-γ ELISpotIL-4 ELISpotiNKT cellsα-Galactosylceramide

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