Production and characterization of monoclonal antibodies against complexes of the NKT cell ligand α-galactosylceramide bound to mouse CD1d

Karl O A Yu, Jin S. Im, Petr A. Illarionov, Rachel M. Ndonye, Amy R. Howell, Gurdyal S. Besra, Steven A. Porcelli

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The α-galactosylceramide (α-GalCer) known as KRN7000 remains the best studied ligand of the lipid-binding MHC class I-like protein CD1d. The KRN7000:CD1d complex is highly recognized by invariant natural killer T (iNKT) cells, an evolutionarily conserved subset of T lymphocytes that express an unusual semi-invariant T cell antigen receptor, and mediate a variety of proinflammatory and immunoregulatory functions. To facilitate the study of glycolipid antigen presentation to iNKT cells by CD1d, we undertook the production of mouse monoclonal antibodies (mAbs) specific for complexes of KRN7000 bound to mouse CD1d (mCD1d) proteins. Three such monoclonal antibodies were isolated that bound only to mCD1d proteins that were loaded with KRN7000 or closely-related forms of α-GalCer. These mAbs showed no reactivity with mCD1d proteins that were not loaded with α-GalCer, nor did they bind to complexes formed by loading mCD1d with the self-glycolipid and putative iNKT cell ligand isoglobotrihexosylceramide. These complex-specific monoclonal antibodies allow the direct detection and monitoring of complexes formed by the binding of KRN7000 and other α-GalCer analogues to mCD1d. The availability of these mAbs should facilitate a wide range of studies on the biology and potential clinical applications of CD1d-restricted iNKT cells.

Original languageEnglish (US)
Pages (from-to)11-23
Number of pages13
JournalJournal of Immunological Methods
Volume323
Issue number1
DOIs
StatePublished - May 31 2007

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Galactosylceramides
Natural Killer T-Cells
Monoclonal Antibodies
Ligands
Glycolipids
Antigen Presentation
T-Lymphocyte Subsets
T-Cell Antigen Receptor
KRN 7000
Lipids
Mouse Cd1d1 protein
Proteins

Keywords

  • Antigen presentation
  • CD1
  • Galactosylceramide
  • Glycolipid
  • Monoclonal antibody
  • NKT cell

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Production and characterization of monoclonal antibodies against complexes of the NKT cell ligand α-galactosylceramide bound to mouse CD1d. / Yu, Karl O A; Im, Jin S.; Illarionov, Petr A.; Ndonye, Rachel M.; Howell, Amy R.; Besra, Gurdyal S.; Porcelli, Steven A.

In: Journal of Immunological Methods, Vol. 323, No. 1, 31.05.2007, p. 11-23.

Research output: Contribution to journalArticle

Yu, Karl O A ; Im, Jin S. ; Illarionov, Petr A. ; Ndonye, Rachel M. ; Howell, Amy R. ; Besra, Gurdyal S. ; Porcelli, Steven A. / Production and characterization of monoclonal antibodies against complexes of the NKT cell ligand α-galactosylceramide bound to mouse CD1d. In: Journal of Immunological Methods. 2007 ; Vol. 323, No. 1. pp. 11-23.
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