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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Gadola, S. Altamirano, M. Brown, D. Dusheiko, G. Klenerman, P. Luzzatto, L. Price, D. A. Woolfson, A. Cerundolo, V. Roberts, I. A. Koezuka, Y. Karadimitris, A. Milstein, C. Chen, J. L. Fersht, A. |
| Description | Author Affiliation: Karadimitris A ( Nuffield Department of Medicine, Institute of Molecular Medicine, Oxford OX3 9DS, United Kingdom.); |
| Abstract | CD1 molecules are specialized in presenting lipids to T lymphocytes, but identification and isolation of CD1-restricted lipid specific T cells has been hampered by the lack of reliable and sensitive techniques. We here report the construction of CD1d-glycolipid tetramers from fully denatured human CD1d molecules by using the technique of oxidative refolding chromatography. We demonstrate that chaperone- and foldase-assisted refolding of denatured CD1d molecules and beta(2)-microglobulin in the presence of synthetic lipids is a rapid method for the generation of functional and specific CD1d tetramers, which unlike previously published protocols ensures isolation of CD1d tetramers loaded with a single lipid species. The use of human CD1d-alpha-galactosylceramide tetramers for ex vivo staining of peripheral blood lymphocytes and intrahepatic T cells from patients with viral liver cirrhosis allowed for the first time simultaneous analysis of frequency and specificity of natural killer T cells in human clinical samples. Application of this protocol to other members of the CD1 family will provide powerful tools to investigate lipid-specific T cell immune responses in health and in disease. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 6 |
| Volume Number | 98 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2001-03-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Antigens, CD1 Metabolism Ceramides Glycolipids Protein Folding Animals Genetics Antigens, CD1d Cells, Cultured Hepatitis C Blood Ligands Liver Cirrhosis Mice Mice, Knockout Oxidation-Reduction Staining And Labeling Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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