Activation and expansion of T-cells using an engineered multivalent signaling platform

• US 7,638,326 B2
• Filed: 06/13/2003
• Issued: 12/29/2009
• Est. Priority Date: 01/03/2002
• Status: Expired due to Term

• Current Assignee: University of Pennsylvania
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Thomas, Anna, June, Carl, Riley, James, Vonderheide, Robert, Maus, Marcela
• Primary Examiner(s): Haddad; Maher M
• Assistant Examiner(s): DAHLE, CHUN WU
• Application Number:
  US10/462,207
• Publication Number:
  US 20040110290A1
• Time in Patent Office: 2,391 Days
• Field of Search: None
• US Class Current: 435/325
• 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 2239/49 Breast
• CPC Class Codes: A61K 2239/58 Prostate
• CPC Class Codes: A61K 39/4615 Dendritic cells
• CPC Class Codes: A61K 39/4622 Antigen presenting cells
• CPC Class Codes: A61K 39/464457 Telomerase or [telomerase reverse transcriptase [TERT]
• 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 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2502/99 genetically modified cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• 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 using an engineered multivalent signaling platform. Compositions of cells activated and expanded by the methods herein are further provided.

Claims

1. An engineered multivalent signaling platform (EMSP) comprising a K562 cell line for use in long term expansion of T cells, wherein said K562 comprises on its surface:

at least one molecule that is capable of stimulating a primary signal to the T cell, wherein said at least one molecule is an exogenous antibody that binds to CD2;

a ligand for CD28, wherein said ligand for CD28 is capable of cross-linking CD28 on T cells and delivering a secondary signal to said T cells, wherein said ligand is selected from the group consisting of anti-CD28 antibody, CD80, and CD86; and

wherein said EMSP is further genetically modified to express the co-stimulatory molecule, 4-1BBL.

2. The EMSP of claim 1 wherein said K562 cell line is a cell that is genetically modified to express a human Fcγ

receptor.

3. The EMSP of claim 2 wherein said human Fcγ

receptor comprises CD32.

4. The EMSP of claim 2 wherein said exogenous antibody is displayed on the surface of said K562 cell line via interaction with the Fcγ

receptor.

5. The EMSP of claim 1 wherein said K562 cell line is a cell that has been modified to express a cytokine.

6. The EMSP of claim 5 wherein said cytokine is selected from the group consisting of IL-2, GM-CSF, IL-4, TNF-α

, and IFN-γ

.

7. The EMSP of claim 1 wherein said T cells comprise CD4⁺ T cells.

8. The EMSP of claim 1 wherein said T cells comprise CD8⁺ T cells.

9. The EMSP of claim 1 wherein said T cells comprise regulatory T cells.

10. The EMSP of claim 2, wherein said anti-CD28 antibody is displayed on the surface of said K562 cell line via interaction with the Fcγ

receptor.

11. The EMSP of claim 4 wherein said antibody is an anti-CD2 antibody.

12. The EMSP of claim 4 wherein said antibody comprises anti-CD2 and anti-CD28.

13. The EMSP of claim 1, further comprising at least an antigen.

14. The EMSP of claim 13 wherein said antigen is selected from the group consisting of a tumor antigen, a bacterial antigen, a fungal antigen, a viral antigen, and an autoantigen.

15. The EMSP of claim 1 wherein said exogenous antibody is anti-CD2 antibody and said ligand for CD28 is anti-CD28 antibody, further wherein said EMSP induces T cells to remain in exponential growth after a third round of stimulation with said EMSP.