US 8,383,554 B2 · Quantitative microarray of intact glycolipid CD1d interaction and correlation with cell-based cytokine production

Abstract

The protein CD1d binds self and foreign glycolipids for presentation to CD1-restricted T cells by means of TCR recognition, and activates TH1 and TH2 chemokines release. Accordingly, a variety of glycolipid ligands were attached to a microarray surface and their binding with CD1d investigated. An α-galactosyl ceramide (α-GalCer) bearing a carbamate group at the 6′-OH position was tethered to the surface and the dissociation constant with CD1d determined. Competition assays were used to determine the dissociation constants (Ki) of the new and intact glycolipids. The para-fluoroheptaphenyl-modified α-GalCer was found to bind most strongly with CD1d (Ki 0.14 μM), two orders of magnitude stronger than α-GalCer and more than three times more selective for IFN-γ release. Various α-GalCer analogs were analyzed and the results showed that the binding affinity of glycolipids to CD1d correlates well with IFN-γ production, but poorly with IL-4 secretion by NKT cells, suggesting that tighter binding ligands could bias cytokine release through the TH1 pathway.

Claims6

Dependent claims

A device comprising:

a plurality of glycan ligands bound to a substrate via a linker bearing a primary amine group suitable for attachment to the substrate, the linker group attached at a derivatized 6′—OH position of the sugar moiety of α-GalCer;
wherein the glycan ligands are α-Galactosylceramide (α-GalCer) derivatives of Formula 1:
wherein R is CHR1—X—Y;
R1 is an organic moiety such as selected from the group consisting of amide, carbamate, carbonate, and urea;
X is alkyl, alkoxyl, or aryl-alkyl;
and
Y is NH2, COOH, N3, alkync, or biotin.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application

12/423,728Filed 14 Apr 2009 · us provisional application

Priority

14 Apr 2008

Published

US 2009/0275483 A15 Nov 2009

Granted

26 Feb 2013

Term ends

Not estimatedThe source records don’t support an estimate.

Classification6 codes

C40B 30/04
Methods of screening libraries: by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand…
C40B 40/12
Libraries containing only organic compounds: libraries containing saccharides or polysaccharides, or derivatives thereof
G01N 2333/70539
Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3: mHC-molecules, e.g. HLA-molecules
G01N 2405/10
Sphingolipids: glycosphingolipids, e.g. cerebrosides, gangliosides

Quantitative microarray of intact glycolipid CD1d interaction and correlation with cell-based cytokine production

Life of this patent

Quantitative microarray of intact glycolipid CD1d interaction and correlation with cell-based cytokine production

Life of this patent

Abstract

Claims6