Generation and characterisation of CD1d tetramer produced by a lentiviral expression system.

Journal article

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.

Authors: Li, D; Chen, N; Mcmichael, A; Screaton, G; Xu, X

• Publication status: Published
• Journal: Journal of immunological methods
• Volume: 330
• Issue: 1-2
• Pages: 57-63
• Publication date: 2008-01-01
• DOI: 10.1016/j.jim.2007.10.018
• EISSN: 1872-7905
• ISSN: 0022-1759
• Language: English
• Keywords: Culture Media; Antigens, CD4; Phenotype; Immunophenotyping; Antigens, CD1; Leukocytes, Mononuclear; Cell Separation; Cell Line; Antigens, CD1d; Killer Cells, Natural; beta 2-Microglobulin; Recombinant Fusion Proteins; Lentivirus; Receptors, Antigen, T-Cell, alpha-beta; Humans; Antigens, CD3; Galactosylceramides; Biotinylation; Cloning, Molecular; Flow Cytometry