Activation and Expansion of T-Cells Using an Engineered Multivalent Signaling Platform as a Research Tool

• US 20140186947A1
• Filed: 12/19/2013
• Published: 07/03/2014
• Est. Priority Date: 01/03/2002
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: June, Carl H., Riley, James L., Maus, Marcela, Thomas, Anna, Vonderheide, Robert
• Granted Patent: US 9,453,199 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• 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 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• 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: A61K 39/464838 Viral antigens
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• 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 2510/04 Immortalised cells
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

Provided are a system and methods for selectively inducing expansion of a population of T cells in the absence of exogenous growth factors, such as lymphokines, and accessory cells for research purposes. The cell based expansion system and methods permit the long-term growth of CTLs, preferably human CTLs. In addition, T cell proliferation can be induced without the need for antigen, thus providing an expanded T cell population that is polyclonal with respect to antigen reactivity. Further provided are methods for using the system and methods to screen and identify antigens related to specific diseases or conditions, tumors, autoimmune disorders, or an infectious disease or pathogen, and to identify target molecule for research purposes, or for developing a vaccine based thereon.

Claims

1-3. -3. (canceled)

4. A method of preparing an aAPC, comprising:

selecting a population of K562 cells and engineering the cells to express Fcγ

receptor CD32, thereby creating K32 cells;

transfecting the K32 cells with the co-stimulatory ligand;

cloning the K32 cells to permit exogenous loading of the anti-CD3 or anti-CD28 antibodies, or a combination thereof, and then loading of the anti-CD3 or anti-CD28 antibodies, or a combination thereof.

5. The method of claim 4, wherein the ligand is 4-1BB (4-1BBL).

6. A system for inducing a population of T cells to rapidly proliferate long term to sufficient numbers for use in research, comprising exposing the population of T cells to an aAPC comprising:

a selected population of K562 cells engineered to express Fcγ

receptor CD32 thereby creating K32 cells, transfected by a co-stimulatory ligand, then coating the K-32-ligand cells with anti-CD3 antibodies or anti-CD28 antibodies, or a combination thereof;

wherein the ligand is 4-1BB (4-1BBL).

7. A system for inducing a population of CD8⁺ T cells to rapidly proliferate long term to sufficient numbers for use in research, comprising exposing the population of T cells to an aAPC comprising:

a selected population of K562 cells engineered to express Fcγ

receptor CD32 thereby creating K32 cells, transfected by a co-stimulatory ligand, then coating the K-32-ligand cells with anti-CD3 antibodies or anti-CD28 antibodies, or a combination thereof;

wherein the ligand is 4-1BB (4-1BBL).

8-97. -97. (canceled)