Abstract
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.
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References
Brennan PJ, Brigl M, Brenner MB (2013) Invariant natural killer T cells: an innate activation scheme linked to diverse effector functions. Nat Rev Immunol 13(1):101–117
Kawano T, Cui J, Koezuka Y et al (1997) CD1d-restricted and TCR-mediated activation of valpha14 NKT cells by glycosylceramides. Science 278(5343):1626–1629
Tessmer MS, Fatima A, Paget C et al (2009) NKT cell immune responses to viral infection. Expert Opin Ther Targets 3(2):153–162
Dhodapkar MV, Geller MD, Chang DH et al (2003) A reversible defect in natural killer T cell function characterizes the progression of premalignant to malignant multiple myeloma. J Exp Med 197(12):1667–1676
Gumperz JE, Miyake S, Yamamura T et al (2002) Functionally distinct subsets of CD1d-restricted natural killer T cells revealed by CD1d tetramer staining. J Exp Med 195(5):625–636
Wilson MT, Johansson C, Olivares-Villagómez D et al (2003) The response of natural killer T cells to glycolipid antigens is characterized by surface receptor down-modulation and expansion. Proc Natl Acad Sci U S A 100(19):10913–10918
Van Der Vliet HJ, Nishi N, Koezuka Y et al (1999) Effects of alpha-galactosylceramide (KRN7000), interleukin-12 and interleukin-7 on phenotype and cytokine profile of human Valpha24+ Vbeta11+ T cells. Immunology 98:557–563
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Kamaladasa, A.I., Malavige, G.N. (2018). Ex Vivo ELISpot Assay to Investigate iNKT Cell Responses in Acute Dengue Infection. In: Kalyuzhny, A. (eds) Handbook of ELISPOT . Methods in Molecular Biology, vol 1808. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8567-8_16
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DOI: https://doi.org/10.1007/978-1-4939-8567-8_16
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-8567-8
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