Methods for altering cell fate to generate T-cells specific for an antigen of interest

• US 7,491,534 B2
• Filed: 04/30/2003
• Issued: 02/17/2009
• Est. Priority Date: 12/22/2000
• Status: Active Grant

• Current Assignee: SAB LLC (W.P. Carey, Inc. (Real Estate))
• Original Assignee: Kirin Holdings Kabushiki Kaisha
• Inventors: Skalhegg, Bjorn Steen, Robl, James M., Collas, Philippe
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US10/426,595
• Publication Number:
  US 20040072288A1
• Time in Patent Office: 2,120 Days
• Field of Search: 435/377, 435/455, 435/357, 435/366, 435/372.3
• US Class Current: 435/377
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/4632 T-cell receptors [TCR]; antibody T-cell receptor constructs
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/464499 Undefined tumor antigens, e.g. tumor lysate or antigens targeted by cells isolated from tumor
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2506/00 Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
• CPC Class Codes: C12N 2517/10 Conditioning of cells for in vitro fecondation or nuclear transfer
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

The invention provides methods for altering the expression profile of a cell to convert the cell from one cell type to a desired cell type (e.g., T-cells). These reprogrammed cells may be used in a variety of medical applications for treating a mammal in need of a particular cell type.

Claims

1. A method of reprogramming a cell to express a T-cell receptor (TCR) reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a permeabilized fibroblast cell with an interphase activated T-cell extract under conditions that result in TCR expression in said permeabilized fibroblast cell; and

(b) selecting a cell that expresses a TCR reactive with an antigen of interest.

2. The method of claim 1, wherein said cell, following step (a), is incubated under conditions that allow the membrane of said cell to reseal.

3. The method of claim 1, wherein said permeabilized fibroblast cell of step (a) is further incubated with RNA or protein for a T-cell receptor reactive with said antigen of interest.

4. The method of claim 1, wherein said method further comprises, prior to step (a), incubating said permeabilized fibroblast cell with a mitotic cell extract under conditions that allow chromatin condensation and nuclear envelope breakdown in said permeabilized fibroblast cell.

5. The method of claim 1, wherein said permeabilized fibroblast cell is a permeabilized human fibroblast cell.

6. The method of claim 1 or 5, wherein said interphase activated T-cell extract is a human interphase activated T-cell extract.

7. The method of claim 1, wherein said permeabilized fibroblast cell is generated by incubating a fibroblast cell with streptolysin O.