Activation and expansion of T cells

• US 7,572,631 B2
• Filed: 03/02/2005
• Issued: 08/11/2009
• Est. Priority Date: 02/24/2000
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Invitrogen Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Berenson, Ronald J, Saund, Zahbea, Bonyhadi, Mark, Kalamasz, Dale, Craig, Stewart, Hardwick, Alan, Law, Che, McMillen, David, Chana, Harjinder Singh
• Primary Examiner(s): Lankford; L Blaine
• Assistant Examiner(s): Kim; Taeyoon
• Application Number:
  US11/070,841
• Publication Number:
  US 20060121005A1
• Time in Patent Office: 1,623 Days
• Field of Search: None
• US Class Current: 435/375
• 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. A method for expanding a population of T cells from a B-cell chronic lymphocytic leukemia (B-CLL) patient by cell surface moiety ligation, comprising:

a. selecting a patient having B-CLL;

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

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

d. 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;

e. 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;

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

g. 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.

2. The method of claim 1 wherein said surface is biocompatible.

3. The method of claim 1 wherein said surface is natural or synthetic.

4. The method of claim 2 wherein the biocompatible surface is biodegradable.

5. The method of claim 2 wherein the biocompatible surface is non-biodegradable.

6. The method of claim 1 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.

7. The method of claim 6 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.