The CD1 family of major histocompatibility complex (MHC)-like molecules specializes in presenting lipid and glycolipid antigens to α/β T lymphocytes, but little is known about the size of the CD1-restricted T cell population or the frequency of T lymphocytes specific for a given glycolipid antigen. Here, we report the generation and use of mouse CD1d1–glycolipid tetramers to visualize CD1d-restricted T cells. In contrast with previous BIAcore-based estimates of very short half-lives for CD1d–glycolipid complexes, we found that the dissociation rate of several different CD1d–glycolipid complexes was very slow. Fluorescent tetramers of mouse CD1d1 complexed with α-galactosylceramide (αGalCer), the antigen recognized by mouse Vα14-Jα281/Vβ8 and human Vα24-JαQ/Vβ11 natural killer T (NKT) cell T cell receptors (TCRs), allowed us for the first time to accurately describe, based on TCR specificity, the entire population of NKT cells in vivo and to identify a previously unrecognized population of NK1.1-negative “NKT” cells, which expressed a different pattern of integrins. In contrast, natural killer (NK) cells failed to bind the tetramers either empty or loaded with αGalCer, suggesting the absence of a CD1d-specific, antigen-nonspecific NK receptor. Mouse CD1d1–αGalCer tetramers also stained human NKT cells, indicating that they will be useful for probing a range of mouse and human conditions such as insulin-dependent diabetes mellitus, tumor rejection, and infectious diseases where NKT cells play an important role.
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5 June 2000
Article|
June 06 1999
In Vivo Identification of Glycolipid Antigen–Specific T Cells Using Fluorescent Cd1d Tetramers
Kamel Benlagha,
Kamel Benlagha
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
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Angela Weiss,
Angela Weiss
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
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Andrew Beavis,
Andrew Beavis
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
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Luc Teyton,
Luc Teyton
bDepartment of Immunology, The Scripps Research Institute, La Jolla, California 92037
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Albert Bendelac
Albert Bendelac
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
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Kamel Benlagha
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
Angela Weiss
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
Andrew Beavis
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
Luc Teyton
bDepartment of Immunology, The Scripps Research Institute, La Jolla, California 92037
Albert Bendelac
aDepartment of Molecular Biology, Princeton University, Princeton, New Jersey 08544
Abbreviations used in this paper: αGalCer, α-galactosylceramide; APC, allophycocyanin; DN, CD4−CD8− double negative; GPI, glycosylphosphatidylinositol; IDDM, insulin-dependent diabetes mellitus; LAM, lipoarabinomannan; NKT cell, natural killer T cell.
Received:
February 17 2000
Revision Requested:
March 24 2000
Accepted:
March 28 2000
Online ISSN: 1540-9538
Print ISSN: 0022-1007
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Exp Med (2000) 191 (11): 1895–1904.
Article history
Received:
February 17 2000
Revision Requested:
March 24 2000
Accepted:
March 28 2000
Citation
Kamel Benlagha, Angela Weiss, Andrew Beavis, Luc Teyton, Albert Bendelac; In Vivo Identification of Glycolipid Antigen–Specific T Cells Using Fluorescent Cd1d Tetramers. J Exp Med 5 June 2000; 191 (11): 1895–1904. doi: https://doi.org/10.1084/jem.191.11.1895
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