The Journal of Experimental Medicine
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Published online 5 June 2000.
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© The Rockefeller University Press, 0022-1007/2000/6/1895/ $5.00
The Journal of Experimental Medicine, Volume 191, Number 11, June 5, 2000 1895-1904

Original Article

In Vivo Identification of Glycolipid Antigen–Specific T Cells Using Fluorescent Cd1d Tetramers

Kamel Benlaghaa, Angela Weissa, Andrew Beavisa, Luc Teytonb, and Albert Bendelaca

a Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544
b Department of Immunology, The Scripps Research Institute, La Jolla, California 92037
Department of Molecular Biology, Princeton, NJ 08544.609-258-2205609-258-5454

abendelac{at}molbio.princeton.edu

The CD1 family of major histocompatibility complex (MHC)-like molecules specializes in presenting lipid and glycolipid antigens to {alpha}/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 {alpha}-galactosylceramide ({alpha}GalCer), the antigen recognized by mouse V{alpha}14-J{alpha}281/Vβ8 and human V{alpha}24-J{alpha}Q/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 {alpha}GalCer, suggesting the absence of a CD1d-specific, antigen-nonspecific NK receptor. Mouse CD1d1–{alpha}GalCer 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.

Key Words: T cell development • T cell receptor • V{alpha}14 NKT cell • natural killer cell • antigen presentation

Abbreviations used in this paper: {alpha}GalCer, {alpha}-galactosylceramide; APC, allophycocyanin; DN, CD4–CD8– double negative; GPI, glycosylphosphatidylinositol; IDDM, insulin-dependent diabetes mellitus; LAM, lipoarabinomannan; NKT cell, natural killer T cell.

© 2000 The Rockefeller University Press


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