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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

DOI

10.1016/j.jim.2007.10.018

Type

Journal article

Journal

J Immunol Methods

Publication Date

31/01/2008

Volume

330

Pages

57 - 63

Keywords

Antigens, CD1, Antigens, CD1d, Antigens, CD3, Antigens, CD4, Biotinylation, 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