Simultaneous stimulation and concentration of cells

• US 6,905,874 B2
• Filed: 02/26/2001
• Issued: 06/14/2005
• Est. Priority Date: 02/24/2000
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Xcyte Therapies, Inc. (Cyclacel Pharmaceuticals Incorporated)
• Inventors: Bonyhadi, Mark, Kalamasz, Dale, Craig, Stewart, Hardwick, Alan, Saund, Narinder, Law, Che, Berenson, Ronald, McMillen, David
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US09/794,230
• Publication Number:
  US 20020058019A1
• Time in Patent Office: 1,569 Days
• Field of Search: 435/325, 435/375, 435/372.3, 436/524
• US Class Current: 435/375
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• 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 39/464499 Undefined tumor antigens, e.g. tumor lysate or antigens targeted by cells isolated from tumor
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2806 against CD2
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2812 against CD4
• CPC Class Codes: C07K 16/2815 against CD8
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/2821 against ICAM molecules, e.g. CD50, CD54, CD102
• CPC Class Codes: C07K 16/2827 against B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2845 against integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: C07K 16/2854 against selectins, e.g. CD62
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 16/2878 : against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/289 : against CD45
• CPC Class Codes: C07K 16/2896 : against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C12N 2500/32 : Amino acids
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• 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 2501/53 : CD2
• CPC Class Codes: C12N 2533/12 : Glass
• CPC Class Codes: C12N 2533/14 : Ceramic
• CPC Class Codes: C12N 2533/18 : Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells
• CPC Class Codes: C12N 2533/30 : Synthetic polymers
• CPC Class Codes: C12N 2533/40 : Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/54 : Collagen; Gelatin
• CPC Class Codes: C12N 2533/70 : Polysaccharides
• CPC Class Codes: C12N 2533/72 : Chitin, chitosan
• CPC Class Codes: C12N 2533/90 : Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0635 : B lymphocytes
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates generally to methods for stimulating cells, and more particularly, to a novel method to concentrate and stimulate cells that maximizes stimulation and/or proliferation of such cells. In the various embodiments, cells are stimulated and concentrated with a surface yielding enhanced proliferation, cell signal transduction, and/or cell surface moiety aggregation. In certain aspects methods for stimulating a population of cells such as T-cells, by simultaneous concentration and cell surface moiety ligation are provided by contacting the population of cells with a surface, that has attached thereto one or more agents that ligate a cell surface moiety and applying a force that predominantly drives cell concentration and cell surface moiety ligation, thereby inducing cell stimulation, cell surface moiety aggregation, and/or receptor signaling enhancement. Also provided are methods for producing phenotypically tailored cells, including T-cells for the use in diagnostics, drug discovery, and the treatment of a variety of indications, including cancer, viral infection, and immune related disorders. Compositions of cells having specific phenotypic properties produced by these processes are further provided.

Claims

1. A method for stimulating a population of T-cells by simultaneous T-cell concentration and cell surface moiety ligation, comprising:

(a) providing a population of cells wherein at least a portion thereof comprises T-cells;

(b) contacting said population of cells with a surface, wherein said surface is a surface of a paramagnetic particle and wherein said surface has attached thereto one or more agents that ligate a cell surface moiety of at least a portion of said T-cells and stimulates at least said portion of T-cells;

(c) applying a magnetic force that predominantly drives T-cell concentration and T-cell surface moiety ligation, thereby inducing T-cell stimulation.

2. The method of claim 1, wherein said surface has attached thereto a first agent that ligates a first cell surface moiety of a T-cell;

and the same or a second surface has attached thereto a second agent that ligates a second moiety of said T-cell, wherein said ligation by the first and second agent induces proliferation of said T-cell.

3. The method of claim 1, wherein said surface is biocompatible.

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

5. The method of claim 3, wherein the biocompatible surface is biodegradable.

6. The method of claim 3, wherein the biocompatible surface is non-biodegradable.

7. The method of claim 3, wherein the biocompatible surface is associated with an implantable device.

8. The method of claim 7, wherein the device is selected from the group consisting of:

a stent, a catheter, a fiber, a hollow fiber, a patch, and a suture.

9. The method of claim 1, wherein the particle is selected from the group consisting of a bead, a microsphere, a nanoparticle, and a colloidal particle.

10. The method of claim 9, wherein said bead is about 5 nanometers to about 500 microns in diameter.

11. The method of claim 1, wherein said agents are independently selected from the group consisting of a protein ligand, a natural ligand, and a synthetic ligand.

12. The method of claim 11, wherein said agents are independently selected from the group consisting of an antibody, an antibody fragment, a peptide, a polypeptide, a glycopeptide, a receptor, a steroid, a hormone, a mitogen, an antigen, a superantigen, a growth factor, a cytokine, a lectin, a viral protein, an adhesion molecule, and a chemokine.

13. The method of claim 12, wherein at least one agent is an antibody or an antibody fragment.

14. The method of claim 12, wherein a first agent is an antibody and a fragment thereof, and a second agent is an antibody or a fragment thereof.

15. The method of claim 14, wherein said first and said second agents are different antibodies.

16. The method of claim 12, wherein said first agent is an anti-CD3 antibody, an anti-CD2 antibody, or an antibody fragment of an anti-CD3 or anti-CD2 antibody.

17. The method of either claim 12, or 15, wherein said second agent is an anti-CD28 antibody or antibody fragment thereof.

18. The method of either claim 12, or 15, wherein said second agent is a natural ligand for CD28.

19. The method of claim 18, wherein said natural ligand comprises B7-1 or B7-2.

20. The method of claim 1, wherein the magnetic force is generated by a magnet having a magnetic field strength ranging from between about 200 gauss to about 12,000 gauss at the surface of the magnet.

21. The method of claim 1, wherein said agents'"'"' attachment to the surface is covalent, noncovalent, electrostatic, or hydrophobic.

22. The method of claim 1, wherein the T-cells that are ligated are separated from the T-cells that are not ligated.

23. The method of claim 1, wherein said T-cells ameliorate immune response dysfunction.

24. A method for stimulation of T-cells by simultaneous cell surface moiety ligation and T-cell aggregation, comprising:

(a) providing a cell population comprising T-cells;

(b) contacting said cell population with a surface, wherein said surface is a surface of a paramagnetic particle and wherein said surface has attached thereto one or more ligands specific for a cell surface moiety;

(c) applying a magnetic force that drives concentration of T-cells and surface; and

(d) incubating said cells for a period of time sufficient to achieve desired stimulation.

25. The method of claim 24, wherein said time sufficient to achieve desired stimulation is from 1 minute to 8 days.

26. The method of claim 25, wherein said time sufficient to achieve desired stimulation is from 1 day to 5 days.

27. The method of claim 24, wherein said surface is biocompatible.

28. The method of claim 27, wherein said surface is natural or synthetic.

29. The method of claim 24, wherein the particle is selected from the group consisting of a bead, a microsphere, a nanoparticle, and a colloidal particle.

30. The method of claim 29, wherein said bead is about 5 nanometers to about 500 microns in diameter.

31. The method of claim 24, wherein said ligand is selected from the group consisting of a protein, a natural ligand, and a synthetic ligand.

32. The method of claim 24, wherein said ligand is selected from the group consisting of an antibody, an antibody fragment, a peptide, a polypeptide, a glycopeptide, a soluble receptor, a steroid, a hormone, a mitogen, an antigen, a ligand, a superantigen, a growth factor, a cytokine, a lectin, and a chemokine.

33. The method of claim 32, wherein at least one ligand is an antibody or a fragment thereof.

34. The method of claim 32, wherein at least two ligands are an antibody or a fragment thereof.

35. The method of claim 32, wherein at least two ligands are present and are different antibodies or fragments thereof.

36. The method of claim 32, wherein at least one ligand is an anti-CD3 antibody, an anti-CD2 antibody, or an antibody fragment of an anti-CD3 or anti-CD2 antibody.

37. The method of either claim 32 or 36, wherein at least one ligand is an anti-CD28 antibody or antibody fragment thereof.

38. The method of either claim 32 or 36, wherein at least one ligand is a natural ligand for CD28.

39. The method of claim 38, wherein said natural ligand comprises B7-1 or B7-2.

40. The method of claim 24, wherein the magnetic force is generated by a magnet having a magnetic field strength ranging from between about 200 gauss to about 12,000 gauss at the surface of the magnet.

41. The method of claim 24, wherein said ligand attachment to the surface is covalent, noncovalent, electrostatic, or hydrophobic.

42. The method of claim 24, further comprising prior to or concurrently with step (d), separating T-cells concentrated with surface from non-concentrated cells.

43. A method adapted for inducing T-cell activation in vivo, comprising providing paramagnetic particles to an animal, said particles having attached thereto, ligands specific for a T-cell surface moiety that induces T-cell activation;

applying a magnetic field to a discrete region of the animal; and

thereby inducing localization and activation of T-cells bound to said particles at said discrete region.