Generation of CD1 tetramers as a tool to monitor glycolipid-specific T cells.
CD1 molecules are beta(2)m-associated HLA class-I-like glycoproteins which have the unique ability to present glycolipid and phospholipid antigens to specific T lymphocytes. To study the biology of CD1 and its role in human disease we developed novel techniques for generation of recombinant CD1/lipid complexes by in vitro refolding. Fluorescent tetrameric complexes made from soluble recombinant CD1d/alpha-galactosylceramide complexes allowed highly sensitive and specific ex vivo and in vitro detection and functional characterization of novel human T-lymphocyte populations. Furthermore, protein crystals were obtained from soluble recombinant CD1b/beta(2)m-proteins loaded either with phosphatidylinositol or ganglioside GM2, which led to the first atomic structure determination of a CD1/lipid complex. The analysis of these crystal structures clarified how CD1b molecules can bind lipid ligands of different size, and revealed a broader spectrum of potential CD1b ligands than previously predicted.
Gadola
Stephan D
SD
Tumour Immunology Unit, The Weatherall Institute of Molecular Medicine, University of Oxford, UK.
Karadimitris
Anastasios
A
Zaccai
Nathan R
NR
Salio
Mariolina
M
Dulphy
Nicolas
N
Shepherd
Dawn
D
Jones
E Yvonne
EY
Cerundolo
Vincenzo
V
D000818:Animals; D018949:Antigens, CD1 / Q000737:chemistry / Q000276:immunology*; D006017:Glycolipids / Q000276:immunology*; D006801:Humans; D020836:Protein Structure, Quaternary; D013601:T-Lymphocytes / Q000276:immunology*
D018949:Antigens, CD1; D006017:Glycolipids