Activation and expansion of cells

• US 20060121005A1
• Filed: 03/02/2005
• Published: 06/08/2006
• Est. Priority Date: 02/24/2000
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Xcyte Therapies, Inc. (Cyclacel Pharmaceuticals Incorporated)
• Inventors: Berenson, Ronald J., Bonyhadi, Mark, Kalamasz, Dale, Craig, Stewart, Hardwick, Alan, Law, Che, Saund, Zahbea, McMillen, David, Chana, Harjinder Singh
• Granted Patent: US 7,572,631 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61K 2239/48 Blood cells, e.g. leukemia or lymphoma
• 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 39/4644 Cancer antigens
• CPC Class Codes: C12N 2501/51 B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
• CPC Class Codes: C12N 2501/515 CD3, T-cell receptor complex
• CPC Class Codes: C12N 2533/00 Supports or coatings for cell culture, characterised by material
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

The present invention relates generally to methods for activating and expanding cells, and more particularly, to a novel method to activate and/or stimulate cells that maximizes the expansion of such cells to achieve dramatically high densities. In the various embodiments, cells are activated and expanded to very high densities in a short period of time. In certain embodiments, cells are activated and expanded to very high numbers of cells in a short period of time. Compositions of cells activated and expanded by the methods herein are further provided.

Claims

1-28. -28. (canceled)

29. A method for activating and expanding a population of T cells by cell surface moiety ligation, comprising:

a. providing a population of cells wherein at least a portion thereof comprises T cells;

b. contacting said population of cells with a surface, wherein said surface has attached thereto one or more agents that ligate a cell surface moiety of at least a portion of the T cells and stimulates said T cells, wherein said surface is present at a ratio of said surface to said cells such that at least one population of antigen-specific T cells is selectively deleted by about 8 days.

30. The method of claim 29 wherein the ratio of surface to cells is 10:

1.

31. The method of claim 29 wherein the ratio of surface to cells is 5:

1.

33. The method of claim 29 or 32 wherein said surface is biocompatible.

34. The method of claim 29 or 32 wherein said surface is natural or synthetic.

35. The method of claim 33 wherein the biocompatible surface is biodegradable.

36. The method of claim 33 wherein the biocompatible surface is non-biodegradable.

37. The method of claim 29 or 32 wherein the first agent is an antibody or an antigen-binding fragment thereof, and the second agent is an antibody or an antigen-binding fragment thereof.

38. The method of claim 37 wherein said first agent is an anti-CD3 antibody or an antigen-binding fragment thereof and said second agent is an anti-CD28 antibody or an antigen-binding fragment thereof.

32. A method for expanding a population of T cells from a B-CLL patient by cell surface moiety ligation, comprising:

a. providing a population of cells from the patient wherein at least a portion thereof comprises T cells;

b. contacting said population of cells for about two hours with a surface, wherein said surface has attached thereto a first agent that ligates a TCR/CD3 complex of a T cell, and a second agent that ligates a CD28 accessory molecule of said T cell;

c. applying a magnetic force that predominantly drives simultaneous T cell concentration and ligation of the TCR/CD3 complex and the CD28 accessory molecule of said T cells;

d. seeding said population of cells at a concentration of between about 0.2×

10⁶ and 5.0×

10⁶ cells/ml in a closed system comprising a disposable bioreactor bag comprising at least one inlet filter and one outlet filter and further contacting said T cells with said surface;

e. perfusing medium through said closed system at about 1 ml/minute starting at about day 8;

f. rocking said bioreactor bag on a rocking platform at about 5-15 rocks/minute;

and wherein said T cells expand to a concentration of about between 15×

10⁶ cells/ml to about 50×

10⁶ cells/ml in less than about two weeks.