Simultaneous stimulation and concentration of cells

• US 6,867,041 B2
• Filed: 04/26/2002
• Issued: 03/15/2005
• 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:
  US10/133,236
• Publication Number:
  US 20030124122A1
• Time in Patent Office: 1,054 Days
• Field of Search: 435/325, 435/326, 435/377, 435/375, 436/526
• US Class Current: 435/377
• 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: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• 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/or 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 cell surface moiety ligation, comprising:

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

(ii) contacting said population of cells with a surface, wherein said surface has attached thereto a first agent that ligates a TCR/CD3 complex of a T-cell, and the same or a second surface has attached thereto a second agent that ligates a CD28 accessory molecule of said T-cell, wherein the ratio of said first agent to said second agent is less than one and wherein said ligation by the first and second agent induces proliferation of said T-cell.

2. The method of claim 1, wherein at least one agent is an antibody or an antibody fragment.

3. The method of claim 1, wherein the first agent is an antibody or a fragment thereof, and the second agent is an antibody or a fragment thereof.

4. The method of claim 1, wherein the first agent is an anti-CD3 antibody, an anti-CD2 antibody, or an antibody fragment of an anti-CD3 or anti-CD2 antibody.

5. The method of claim 1, wherein the second agent is an anti-CD28 antibody or antibody fragment thereof.

6. The method of claim 1, wherein the first agent is an anti-CD3 antibody and the second agent is an anti-CD28 antibody.

7. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

30 to about 1;

100.

8. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

30.

9. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

25.

10. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

20.

11. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

15.

12. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

10.

13. The method of claim 6, wherein the anti-CD3 antibody and the anti-CD28 antibody are present at a ratio of about 1:

3.

14. The method of claim 1, wherein the first agent is an anti-CD3 antibody and the second agent is a ligand for CD28.

15. The method of claim 14, wherein the ligand is a natural ligand for CD28.

16. The method of claim 15, wherein the natural ligand is B7.

17. A method for stimulating a population of T-cells by cell surface moiety ligation, comprising:

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

(ii) contacting said population of cells with a surface, wherein said surface has attached thereto a first agent that ligates a TCR/CD3 complex of a T-cell, and the same or a second surface has attached thereto a second agent that ligates a CD28 accessory molecule of said T-cell, wherein the final ratio of said surface to said cells is greater than 1 on day 1 and wherein additional surface is added sequentially to the cells thereafter for up to 10 days at a final ratio of surface to cells of about 1;

5, and wherein said ligation by the first and second agent induces proliferation of said T-cell.

18. The method of claim 17, wherein said final ratio is about 1:

1 surface;

cells on day 1 and wherein additional surface is added sequentially to the cells thereafter every day for up to 10 days at a final ratio of about 1;

5.

19. The method of claim 17, wherein said final ratio is about 3:

1 surface;

cells on day 1 and at a final ratio of about 1;

5 on days 3 and 5.

20. The method of claim 17, wherein said final ratio is about 2:

1 surface;

cells on day 1 and at a final ratio of about 1;

5 on days 3 and 5.

21. The method of claim 21, wherein said final ratio is about 1:

1 surface;

cells on day 1 and at a final ratio of about 1;

5 on days 3 and 5.