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The alpha-galactosylceramide (alphaGalCer)-loaded CD1d tetramer remains the most powerful tool in identifying natural killer T (NKT) cells, a subpopulation of T cells that express an unusual semi-invariant T cell antigen receptor, and mediate a variety of proinflammatory and immunoregulatory functions. The difficulty of generating large amounts of the alphaGalCer-CD1d tetramer has limited its availability and consequently hampered the study of NKT cells. In this report, we used a lentiviral system to generate stable cell lines producing beta2m-CD1d single chain protein in large quantities and in a relatively short period of time. When the protein was loaded with alphaGalCer and tetramerised with fluorescence-labelled streptavidin, its ability to efficiently bind to NKT cells was confirmed both by phenotype analysis and functional study. The CD1d tetramer generated from these stable cell lines should facilitate a wide range of studies on the biology and clinical applications of CD1d-restricted NKT cells.

Original publication




Journal article


J Immunol Methods

Publication Date





57 - 63


Antigens, CD1, Antigens, CD1d, Biotinylation, CD3 Complex, CD4 Antigens, Cell Line, Cell Separation, Cloning, Molecular, Culture Media, Flow Cytometry, Galactosylceramides, Humans, Immunophenotyping, Killer Cells, Natural, Lentivirus, Leukocytes, Mononuclear, Phenotype, Receptors, Antigen, T-Cell, alpha-beta, Recombinant Fusion Proteins, beta 2-Microglobulin