Methods for selectively stimulating proliferation of T cells

• US 6,534,055 B1
• Filed: 05/04/1995
• Issued: 03/18/2003
• Est. Priority Date: 11/23/1988
• Status: Expired due to Term

• Current Assignee: Genetics Institute LLC (Pfizer Inc.), the united states of america as represented by the secretary of the navy, University of Alabama (University of Alabama System), Board of Regents of the University of Michigan (University of Michigan)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Nabel, Gary J., June, Carl H., Thompson, Craig B., Gray, Gary S., Rennert, Paul D.
• Primary Examiner(s): Gambel, Phillip
• Application Number:
  US08/435,816
• Time in Patent Office: 2,875 Days
• Field of Search: 435/2, 435/240.2, 435/7.1, 435/375, 424/130.1, 424/141.1, 424/143.1, 424/144.1, 424/153.1, 424/154.1, 424/173.1, 424/93.7, 424/93.71, 424/534, 424/577, 424/578, 530/387.1, 530/388.1, 530/388.2, 530/388.22, 530/388.7, 530/388.73, 530/388.75
• US Class Current: 424/93.71
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 39/464838 Viral antigens
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 14/70532 B7 molecules, e.g. CD80, CD86
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• 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 2740/16011 Human Immunodeficiency Virus, HIV
• CPC Class Codes: C12N 2795/14111 Inoviridae
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

Methods for inducing a population of T cells to proliferate by activating the population of T cells and stimulating an accessory molecule on the surface of the T cells with a ligand which binds the accessory molecule are described. T cell proliferation occurs in the absence of exogenous growth factors or accessory cells. T cell activation is accomplished by stimulating the T cell receptor (TCR)/CD3 complex or the CD2 surface protein. To induce proliferation of an activated population T cells, an accessory molecule on the surface of the T cells, such as CD28, is stimulated with a ligand which binds the accessory molecule. The T cell population expanded by the method of the invention can be genetically transduced and used for immunotherapy or can be used in methods of diagnosis.

Claims

1. A method for expanding a population of CD4+ T cells in number of from about 100 to about 100,000 fold over the original CD4+ T cell population or to about 10 log₁₀ to 12 log₁₀, comprising:

a) contacting a population of CD4+ T cells with a solid phase surface comprising

1) an anti-CD3 antibody; and

2) a second agent which stimulates a CD28-associated signal in the T cells selected from a group consisting of an anti-CD28 antibody, B7-1, and B7-2, wherein said anti-CD3 antibody and said second agent are immobilized on the same solid phase surface;

b) adding exogenous IL-2 to the population of CD4+ T cells, such that the population of CD4+ T cells increase in number from about 100 to about 100,000 fold over the original CD4+ T cell population or to about 10 log₁₀ to 12 log₁₀.

2. The method of claim 1, wherein the anti-CD3 antibody is the monoclonal antibody OKT3.

3. The method of claim 1, wherein the second agent is an anti-CD28 monoclonal antibody.

4. The method of claim 1, wherein IL-2 is added to the population of CD4⁺ T cells in an amount sufficient to maintain proliferation of the CD4⁺ T cells.

5. The method of claim 1, wherein IL-2 is added to the population of CD4⁺ T cells from the beginning of the culture and supplemented every other day in an amount sufficient to maintain proliferation of the CD4⁺ T cells.

6. The method of claim 1, wherein the anti-CD3 antibody is a momoclonal antibody.

7. The method of claim 1, wherein the anti-CD-28 antibody is a monoclonal antibody.

8. A method for preparing a renewable source of CD4⁺ T cells, comprising expanding a population of CD4⁺ T cells according to the method of claim 1 and storing a portion of the cells under conditions which will not significantly affect their viability upon thawing.

16. The method of claim 1, wherein said anti-CD3 antibody and said anti-CD28 antibody are whole antibodies.

18. The method of claim 1, wherein said solid phase surface is a bead.

19. The method of claim 18, wherein the bead is a magnetic immunobead.

22. The method of claim 1, wherein said solid phase surface is a tissue culture dish.

24. The method of claim 1, wherein the anti-CD3 antibody and the second agent are immobilized on the solid phase surface via a covalent modification.

26. The method of claim 1, wherein the anti-CD3 antibody and the second agent are immobilized on the solid phase surface via an avidin-biotin complex.

28. The method of claim 1, wherein the anti-CD3 antibody and the second agent are directly immobilized on the solid phase surface.

9. A method for expanding a population of T cells in number of from about 100 to about 100,000 fold over the original T cell population or to about 10 log₁₀ to 12 log₁₀, comprising:

a) contacting a population of T cells with a solid phase surface comprising

1) an anti-CD3 antibody; and

2) a second agent which stimulates a CD28-associated signal in the T cells selected from the group consisting of an anti-CD28 antibody, B7-1, and B7-2, wherein said anti-CD3 antibody and said second agent are immobilized on the same solid phase surface;

b) adding exogenous IL-2 to the population of T cells, such that the population of T cells increase in number from about 100 to about 100,000 fold over the original T cell population or to about 10 log₁₀ to 12 log₁₀.

10. The method of claim 9, wherein the anti-CD3 antibody is the monoclonal antibody OKT3.

11. The method of claim 9, wherein the second agent is an anti-CD28 monoclonal antibody.

12. The method of claim 9, wherein IL-2 is added to the population of T cells in an amount sufficient to maintain proliferation of the T cells.

13. The method of claim 9, wherein IL-2 is added to the population of T cells from the beginning of the culture and supplemented every other day in an amount sufficient to maintain proliferation of the T cells.

14. The method of claim 9, wherein the anti-CD3 antibody is a monoclonal antibody.

15. The method of claim 9, wherein the anti-CD28 antibody is a monoclonal antibody.

17. The method of claim 9, wherein said anti-CD3 antibody and said anti-CD28 antibody are whole antibodies.

20. The method of claim 9, wherein said solid phase surface is a bead.

21. The method of claim 20, wherein the bead is a magnetic immunobead.

23. The method of claim 9, wherein said solid phase surface is a tissue culture dish.

25. The method of claim 9, wherein the anti-CD3 antibody and the second agent are immobilized on the solid phase surface via a covalent modification.

27. The method of claim 9, wherein the anti-CD3 antibody and The second agent are immobilized on the solid phase surface via an avidin-biotin complex.

29. The method of claim 9, wherein the anti-CD3 antibody and the second agent are directly immobilized on the solid phase surface.