US 7,022,483 B1Granted

Methods for identifying and isolating antigen-specific T cells

AssigneeSalvatore Albani

InventorSalvatore Albani

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Abstract

The present invention is directed to methods for the identification and isolation of antigen-specific T cells using a novel class of artificial antigen presenting cells. The resulting T cells may be used to produce expanded T cell populations as well as for modulating T cell responses. In general, the artificial antigen presenting cells useful in such methods are liposomes that contain MHC:peptide complexes presented on the outer surface of the liposome. Such artificial antigen presenting cells may also include accessory molecules, co-stimulatory molecules, adhesion molecules, and other molecules irrelevant to T cell binding or modulation that are used in the binding of artificial antigen presenting cells to solid support systems that may be used in the retrieval and identification of antigen-specific T cells.

Claims9

Dependent claims

A method of identifying a T cell specific for an antigen of interest, comprising:

a) contacting a biological sample containing T cells specific for an antigen of interest with an artificial antigen presenting cell that presents a peptide derived from the antigen of interest in order to form a complex comprised of a T cell specific for the antigen of interest and an artificial antigen presenting cell that presents the peptide derived from the antigen of interest, wherein the artificial antigen presenting cell comprises:
i. a liposome comprising a lipid bilayer, wherein the lipid bilayer is comprised of neutral phospholipids and cholesterol;
ii. at least one GM-1 ganglioside molecule disposed in the lipid bilayer;
iii. a cholera toxin β subunit bound to one of the GM-1 ganglioside molecules;
iv. an MHC molecule loaded with the peptide derived from the antigen of interest, wherein the antigen-loaded MHC molecule is bound to the cholera toxin β subunit;
and
v. an accessory molecule that can stabilize an interaction between a T cell receptor and the antigen-loaded MHC molecule;
and
b) detecting the complex, if formed, thereby identifying a T cell specific for the antigen of interest.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application: 09/421,506Filed 19 Oct 1999 · us provisional application
Priority: 20 Oct 1998
Granted: 4 Apr 2006
Term ends: Not estimatedThe source records don’t support an estimate.

Classification27 codes

G01N 33/56977
Animal cells: hLA or MHC typing
A61K 9/127
Dispersions; Emulsions: synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
A61K 40/11
Characterised by the cell type used: t-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer…
A61K 40/22
Characterised by the effect or the function of the cells: immunosuppressive or immunotolerising