Novel Artificial Antigen Presenting Cells and Uses Thereof

• US 20140212446A1
• Filed: 04/08/2014
• Published: 07/31/2014
• Est. Priority Date: 05/27/2004
• 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: Riley, James L., June, Carl H., Vonderheide, Robert H., Aqui, Nicole, Suhoski, Megan M.
• Granted Patent: US 9,555,105 B2
• Time in Patent Office: Days
• US Class Current: 424/185.1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2039/5154 Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 2039/55522 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 2039/55527 Interleukins
• CPC Class Codes: A61K 2039/55533 IL-2
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/21 Retroviridae, e.g. equine infectious anemia virus
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 47/642 the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/2302 : Interleukin-2 (IL-2)
• 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 2502/1114 : T cells
• CPC Class Codes: C12N 2502/99 : genetically modified cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2740/15043 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0634 : Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0635 : B lymphocytes
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 5/0637 : Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg
• CPC Class Codes: C12N 5/0639 : Dendritic cells, e.g. Langherhans cells in the epidermis
• CPC Class Codes: C12N 5/0694 : Cells of blood, e.g. leukemia cells, myeloma cells
• CPC Class Codes: G01N 33/505 : involving T-cells
• CPC Class Codes: G01N 33/56972 : White blood cells

Abstract

The invention relates to novel artificial antigen presenting cells (aAPCs). The aAPC comprises at least one stimulatory ligand and at least one co-stimulatory ligand where the ligands each specifically bind with a cognate molecule on a T cell of interest, thereby mediating expansion of the T cell. The aAPC of the invention can further comprise additional molecules useful for expanding a T cell of interest. The aAPC of the invention can be used as an “off the shelf” APC that can be readily designed to expand a T cell of interest. Also, the aAPC of the invention can be used identify the stimulatory, co-stimulatory, and any other factors that mediate growth and expansion of a T cell of interest. Thus, the present invention provides powerful tools for development of novel therapeutics where activation and expansion of a T cell can provide a benefit.

Claims

1. An isolated artificial antigen presenting cell (aAPC), said aAPC comprising a K562 cell transduced using a lentiviral vector (LV), wherein said LV comprises a nucleic acid encoding at least one immune stimulatory ligand and at least one co-stimulatory ligand and further wherein said aAPC expresses said stimulatory ligand and said co-stimulatory ligand and can stimulate and expand a T cell contacted with said aAPC.

2. The isolated aAPC of claim 1, wherein said stimulatory ligand is a polypeptide selected from the group consisting of a major histocompatibility complex Class I (MHC class I) molecule loaded with an antigen, an anti-CD3 antibody, an anti-CD28 antibody, and an anti-CD2 antibody.

3. The isolated aAPC of claim 1, wherein said co-stimulatory ligand is at least one co-stimulatory ligand selected from the group consisting of CD7, B7-1 (CD80), B7-2 (CD86), PD-L1, PD-L2, 4-1BBL, OX40L, ICOS-L, ICAM, CD30L, CD40, CD70, CD83, HLA-G, MICA, MICB, HVEM, lymphotoxin beta receptor, ILT3, ILT4, 3/TR6, and a ligand that specifically binds with B7-H3.

4. The isolated aAPC of claim 1, wherein said co-stimulatory ligand specifically binds with at least one of a co-stimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, LFA-1, CD2, CD7, LIGHT, NKG2C, B7-H3, BTLA, Toll ligand receptor and a ligand that specifically binds with CD83.

5. The isolated aAPC of claim 1, wherein said co-stimulatory ligand is an antibody that specifically binds with at least one molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, LFA-1, CD2, CD7, LIGHT, NKG2C, B7-H3, Toll ligand receptor and a ligand that specifically binds with CD83.

6. The isolated aAPC of claim 1, wherein said aAPC further comprises Fcγ

receptor selected from the group consisting of a CD32 molecule and a CD64 molecule.

7. The isolated aAPC of claim 1, wherein said LV comprises a nucleic acid encoding at least one antigen selected from the group consisting of a tumor antigen, a viral antigen, a bacterial antigen, a peptide-MHC tetramer, a peptide-MHC trimer, a peptide-MHC dimer, and a peptide-MHC monomer.

8. The isolated aAPC of claim 7, wherein said tumor antigen is selected from the group consisting of MAGE-1, MAGE-2, MAGE-3, MART-1, GP100, CEA, HER-2/Neu, PSA, WT-1, MUC-1, MUC-2, MUC-3, MUC-4, and telomerase.

9. The isolated aAPC of claim 1, wherein said LV comprises a nucleic acid encoding at least one peptide selected from a cytokine and a chemokine.

10. The isolated aAPC of claim 9, wherein said cytokine is at least one cytokine selected from the group consisting of IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-15, IL-21, interferon-alpha (IFNα

), interferon-beta (IFNβ

), interferon-gamma (IFNγ

), tumor necrosis factor-alpha (TNFα

), tumor necrosis factor-beta (TNFβ

), granulocyte macrophage colony stimulating factor (GM-CSF), and granulocyte colony stimulating factor (GCSF).

11. A method for specifically inducing proliferation of a T cell expressing a known co-stimulatory molecule, said method comprising contacting said T cell with an aAPC of claim 1, further wherein said co-stimulatory ligand specifically binds with said known co-stimulatory molecule, thereby specifically inducing proliferation of said T cell.

12. A method for specifically inducing proliferation of a T cell expressing a known co-stimulatory molecule, said method comprising contacting a population of T cells comprising at least one T cell expressing said known co-stimulatory molecule with an aAPC of claim 1, wherein said aAPC expresses at least one co-stimulatory ligand that specifically binds with said known co-stimulatory molecule, wherein binding of said known co-stimulatory molecule with said co-stimulatory ligand induces proliferation of said T cell.

13. A method of specifically expanding a T cell population subset, said method comprising contacting a population of T cells comprising at least one T cell of said subset with an aAPC of claim 1, wherein said aAPC comprises at least one co-stimulatory ligand that specifically binds with a co-stimulatory molecule on said T cell of said subset, wherein binding of said co-stimulatory molecule with said co-stimulatory ligand induces proliferation of said T cell of said subset, thereby specifically expanding a T cell population subset.

14. A method of identifying a co-stimulatory ligand, or combination thereof, that specifically induces activation of a T cell subset, said method comprising contacting a population of T cells with an aAPC of claim 1, and comparing the level of proliferation of said T cell population with the level of proliferation of an otherwise identical population of T cells not contacted with said aAPC, wherein a greater level of proliferation of said T cells contacted with said aAPC compared with the level of proliferation of said otherwise identical population of T cells not contacted with said aAPC, is an indication that said co-stimulatory ligand specifically induces activation of said T cell.

15. A method for inducing a T cell response to an antigen in a mammal, said method comprising administering to said mammal the aAPC of claim 1, wherein said aAPC further comprises an MHC Class I molecule loaded with said antigen, wherein said aAPC induces proliferation of a T cell specific for said antigen, thereby inducing a T cell response to said antigen in said mammal.

16. A method of inducing a T cell response to an antigen in a mammal in need thereof, said method comprising obtaining a population of cells from said mammal wherein said population comprises T cells, contacting said population of cells with an aAPC of claim 1, wherein said aAPC further comprises an MHC Class I complex loaded with said antigen, whereby contacting said cells with said aAPC induces proliferation of an antigen-specific T cell specific for said antigen, isolating said antigen-specific T cell from said population of cells, and administering said antigen-specific T cells to said mammal, thereby inducing a T cell response to said antigen in said mammal.

17. A method of specifically expanding a population of T regulatory (Treg) cells, the method comprising contacting said population with an aAPC of claim 1, wherein said aAPC further comprises an Fcγ

receptor loaded with an anti-CD3 antibody and an anti-CD28 antibody, the method further comprising contacting said population of cells with a cytokine, wherein binding of said anti-CD3 antibody and said anti-CD28 antibody with said Treg cells induces proliferation of said Treg cells, thereby specifically expanding a population of Treg cells.

18. The method of claim 17, wherein said cytokine is interleukin-2.

19. A kit for specifically inducing proliferation of a T cell expressing a known co-stimulatory molecule, said kit comprising an effective amount of an aAPC, wherein said aAPC comprises a K562 cell transduced using a lentiviral vector (LV), wherein said LV comprises a nucleic acid encoding at least one co-stimulatory ligand that specifically binds said known co-stimulatory molecule, wherein binding of said known co-stimulatory molecule with said co-stimulatory ligand stimulates and expands said T cell, said kit further comprising an applicator and an instructional material for the use of said kit.

20. A kit for specifically inducing proliferation of a T cell expressing a known stimulatory molecule, said kit comprising an effective amount of an aAPC, wherein said aAPC comprises a K562 cell transduced using a lentiviral vector (LV) wherein said LV comprises a nucleic acid encoding at least one stimulatory ligand that specifically binds said known stimulatory molecule, wherein binding of said known stimulatory molecule with said stimulatory ligand stimulates and expands said T cell, said kit further comprising an applicator and an instructional material for the use of said kit.

21. A kit for specifically expanding a T cell population subset, said kit comprising an effective amount of an aAPC, wherein said aAPC comprises a K562 cell transduced using a lentiviral vector (LV), wherein said LV comprises a nucleic acid encoding at least one co-stimulatory ligand that specifically binds a co-stimulatory molecule on said T cell population, wherein binding of said co-stimulatory molecule with said co-stimulatory ligand stimulates and expands said T cell population, said kit further comprising an applicator and an instructional material for the use of said kit.

22. A kit for identifying a co-stimulatory ligand, or combination of said ligands, that specifically induces activation of a T cell subset, said kit comprising a plurality of aAPCs wherein each said aAPC comprises a K562 cell transduced using a lentiviral vector (LV), wherein said LV comprises a nucleic acid encoding at least one known co-stimulatory ligand that specifically binds with a co-stimulatory molecule, said kit further comprising an applicator and an instructional material for the use of said kit.