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

By K. Benlagha, A. Weiss, A. Beavis, L. Teyton, A. Bendelac. Journal of Experimental Medicine, 2000

604 citations recorded by the source corpus.

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.

Subfields: Immunology, Oncology

View Paper: https://www.semanticscholar.org/paper/15d9a06efbba28bc9f3a40e728913756b761debb

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In Immunology

1. M. Kronenberg
2. D. Godfrey
3. A. Bendelac
4. L. Teyton
5. M. Brenner
6. L. Gapin
7. K. Hoebe
8. B. Beutler

In Oncology

1. M. Kronenberg
2. A. Bendelac
3. L. Teyton
4. L. Gapin
5. M. Brenner
6. K. Garcia
7. Alan R. Fersht

Citations in the corpus

• The transcription factor PLZF (Zbtb16) directs the effector program of the NKT cell lineage (2008, methodology): “Flow Cytometry CD1d-aGalCer tetramers were prepared as previously described (Benlagha et al., 2000).”
• The Paradox of Immune Molecular Recognition of α-Galactosylceramide: Low Affinity, Low Specificity for CD1d, High Affinity for αβ TCRs1 (2003, methodology): “Recombinant soluble murine CD1d molecules were produced in a fly expression system as previously described (12).”
• The crystal structure of human CD1d with and without α-galactosylceramide (2005, background): “The glycolipid α-galactosylceramide (α-GalCer), isolated from a marine sponge, binds human CD1d and mouse CD1d with high affinity and specifically activates the main population of CD1d-restricted T cells called NKT cell”
• Going both ways: immune regulation via CD1d-dependent NKT cells. (2004, background): “org Volume 114 Number 10 November 2004 fluorochrome-conjugated, tetrameric complexes of CD1d loaded with α-GalCer (9, 20, 21).”
• Vα24-JαQ-Independent, CD1d-Restricted Recognition of α-Galactosylceramide by Human CD4+ and CD8αβ+ T Lymphocytes1 (2002, methodology): “For these reasons, it has been speculated that the V -chain does not contribute to the specific recognition of CD1d- GC (21).”
• The Vα14 NKT Cell TCR Exhibits High-Affinity Binding to a Glycolipid/CD1d Complex1 (2002, methodology): “It is noteworthy that the NKT cell TCR affinity for -GalCer-loaded CD1d molecules is higher that the affinity of any other published experimental Ags for CD4 and CD8 T cells (29, 30).”
• The unconventional lifestyle of NKT cells (2002, background): “cgi?/&Q=cd8a%20or%20cd8b&ORG=Mm">CD8 — they are either CD4 + or double negative (DN”
• Natural killer T cells recognize diacylglycerol antigens from pathogenic bacteria (2006, methodology): “We evaluated V α 14 i NKT cell activation by measuring CD25 and CD69 on the surface of V α 14 i NKT cells, which bind to CD1d tetramers loaded with α-GalCe”
• Distinct Functional Lineages of Human Vα24 Natural Killer T Cells (2002, methodology): “1 cells with respect to their pattern of integrins (15).”
• Functionally Distinct Subsets of CD1d-restricted Natural Killer T Cells Revealed by CD1d Tetramer Staining (2002, background): “Most tetramer positive T cells were positive for CD161 (mean 72 14%).”