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Semi-invariant natural killer T (iNKT) cells are CD1d-restricted innate-like lymphocytes that recognize lipid agonists. Activated iNKT cells have immunoregulatory properties. Human and mouse iNKT cell functions elicited by different glycolipid agonists are highly conserved, making the mouse an excellent animal model for understanding iNKT cell biology in vivo. This unit describes basic methods for the characterization and quantification (see Basic Protocol 1) and functional analysis of mouse iNKT cells in vivo or in vitro. This unit also contains protocols that describe enrichment and purification of iNKT cells, generation of CD1d tetramer, and lipid antigen loading onto cell-bound and soluble CD1d for activation of NKT cell hybridomas. © 2017 by John Wiley & Sons, Inc.

Original publication




Journal article


Curr Protoc Immunol

Publication Date





14.13.1 - 14.13.55


CD1d, CD1d tetramers, NKT cells, T cells, cells, cytokines, immune regulation, innate immunity, lipid agonists, murine iNKT, self ligands, Animals, Antigens, Surface, Biomarkers, Flow Cytometry, Immunophenotyping, Lymphocyte Activation, Mice, Natural Killer T-Cells, Phenotype, T-Lymphocyte Subsets