Activation and expansion of T-cells using an engineered multivalent signaling platform as a research tool

• US 8,637,307 B2
• Filed: 05/10/2010
• Issued: 01/28/2014
• Est. Priority Date: 01/03/2002
• Status: Expired due to Term

• 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
• Primary Examiner(s): DAHLE, CHUN WU
• Application Number:
  US12/777,053
• Publication Number:
  US 20100261269A1
• Time in Patent Office: 1,359 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• 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. An isolated population of T cells expanded and restimulated by a method comprising:

contacting a population of T cells with an engineered K562 cell, said engineered K562 cell comprising on its surface;

     1) an exogenous antibody that binds to CD3 or CD2,

     2) a molecule that binds CD28, wherein the molecule that binds CD28 is selected from the group consisting of anti-CD28 antibody, CD80, and CD86, and

     3) the natural 4-1BBL; and

expanding said population of T cells by culturing said population of T cells under conditions and time sufficient to induce cell division, wherein said contacting and/or said culturing occurs in the absence of exogenously added cytokines, wherein said T cells so expanded are substantially free of said engineered K562 cell, followed by restimulating said T cells wherein said T cells remain in exponential growth after restimulation.

3. The method of claim 1, wherein said 4-1BBL is full length 4-1BBL.

2. An isolated population of T cells expanded and restimulated by a method comprising:

contacting a population of T cells with an engineered K562 cell, said engineered K562 cell comprising on its surface;

     1) an exogenous antibody that binds to CD3 or CD2,

     2) a molecule that binds CD28, wherein the molecule that binds CD28 is selected from the group consisting of anti-CD28 antibody, CD80, and CD86, and

     3) the natural 4-1BBL; and

expanding said population of T cells by culturing said T cells under conditions and time sufficient to induce cell division, wherein said T cells so expanded are substantially free of said engineered K562 cell, followed by restimulating said T cells wherein said T cells remain in exponential growth after restimulation.

4. The method of claim 2, wherein said 4-1BBL is full length 4-1BBL.