Methods for selectively stimulating proliferation of T cells

• US 5,858,358 A
• Filed: 06/03/1994
• Issued: 01/12/1999
• Est. Priority Date: 04/07/1992
• Status: Expired due to Term

• Current Assignee: Dana-Farber Cancer Institute, Genetics Institute LLC (Pfizer Inc.), the united states of america as represented by the secretary of the navy, Board of Regents of the University of Michigan (University of Michigan)
• Original Assignee: Dana-Farber Cancer Institute, the united states of america as represented by the secretary of the navy, Board of Regents of the University of Michigan (University of Michigan)
• Inventors: Freeman, Gordon J., Nabel, Gary J., June, Carl H., Thompson, Craig B., Gray, Gary S., Rennert, Paul D.
• Primary Examiner(s): Eisenschenk, Frank C.
• Application Number:
  US08/253,751
• Time in Patent Office: 1,684 Days
• Field of Search: 530/387.1, 530/388.12, 530/388.7, 530/388.75, 424/130.1, 424/143.1, 424/154.1, 435/383
• US Class Current: 424/130.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• 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 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 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• 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: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/74 Inducing cell proliferation
• 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

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 or CD9, 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 inducing a population of CD8⁺ T cells to proliferate, comprising:

a) activating a population of T cells; and

b) stimulating a CD9 antigen on the surface of the T cells with a ligand which binds the CD9 antigen, the activating and stimulating steps thereby inducing proliferation of the T cells.

2. The method of claim 1, wherein the population of T cells is activated by contacting the T cells with an anti-CD3 antibody.

3. The method of claim 2, wherein anti-CD3 antibody is an anti-human CD3 monoclonal antibody.

4. The method of claim 3, wherein the anti-CD3 antibody is immobilized on a solid phase surface.

5. The method of claim 1, wherein the population of T cells is activated by contacting the T cells with an anti-CD2 antibody.

6. The method of claim 1, wherein the population of T cells is activated by contacting the T cells with a protein kinase C activator and a calcium ionophore.

7. The method of claim 1, wherein the ligand binds a peptide comprising an amino acid sequence

(Xaa₁)_n -Gly-Xaa₂ -Trp-Leu-Xaa₃ -Xaa₄ -Asp(Glu)-(Xaa₅)_n (SEQ ID NO:

5),

wherein Xaa₄ may or may not be present, Xaa₁, Xaa₂, Xaa₃, Xaa₄ and Xaa₅ are any amino acid residue and n=0-20.

8. The method of claim 7, wherein Xaa₂ is Cys, Ile or Leu, Xaa₃ is Leu or Arg and Xaa₄, if present, is Arg, Pro or Phe.

9. The method of claim 2, wherein the ligand is a monoclonal antibody ES5.2D8 or portions thereof.

10. The method of claim 1, further comprising contacting the T cells with an antigen or portion thereof.

11. The method of claim 1, further comprising

c) monitoring proliferation of the T cells in response to continuing exposure to the ligand;

and

d) reactivating and restimulating the T cells when the rate of T cell proliferation has decreased to induce further proliferation of the T cells.

12. The method of claim 11, further comprising repeating the steps (c)-(d) to produce a population of T cells increased in number of from about 100- to about 100,000-fold the original T cell population.

13. A method for stimulating a population of CD8⁺ T cell to proliferate, comprising

a) contacting a population of T cells with

(1) a first agent which stimulates a TCR/CD3 complex-associated signal in the T cells;

and

(2) a second agent which stimulates a CD9 antigen on the surface of the T cells.

14. The method of claim 13, wherein the first agent is an anti-CD3 antibody.

15. The method of claim 14, wherein anti-CD3 antibody is an anti-human CD3 monoclonal antibody.

16. The method of claim 15, wherein the anti-CD3 antibody is immobilized on a solid phase surface.

17. The method of claim 13, wherein the second agent is a ligand which binds a peptide comprising an amino acid sequence

(Xaa₁)_n -Gly-Xaa₂ -Trp-Leu-Xaa₃ -Xaa₄ -Asp(Glu)-(Xaa₅)_n (SEQ ID NO:

5),

wherein Xaa₄ may or may not be present, Xaa₁, Xaa₂, Xaa₃, Xaa₄ and Xaa₅ are any amino acid residue and n =0-20.

18. The method of claim 17, wherein Xaa₂ is Cys, Ile or Leu, Xaa₃ is Leu or Arg and Xaa₄, if present, is Arg, Pro or Phe.

19. The method of claim 14, wherein the second agent is a monoclonal antibody ES5.2D8.

20. The method of claim 14, further comprising:

b) separating the anti-CD3 antibody from the T cells and second agent;

c) monitoring proliferation of the T cells in response to continuing exposure to the second agent; and

d) restimulating the T cells with the anti-CD3 antibody and the second agent when the rate of T cell proliferation has decreased to induce further proliferation of the T cells.

21. The method of claim 20, further comprising repeating steps (b)-(d) to produce a population of T cells increased in number of from about 100- to about 100,000-fold the original T cell population.

22. The method of claim 20, wherein the population of CD8⁺ T cells is tumor infiltrating lymphocytes obtained from an individual afflicted with cancer and the method further comprises restoring the T cells to the individual.

23. The method of claim 22, further comprising genetically transducing the T cells and restoring the transduced T cells to an individual.

24. A substantially homogeneous CD8⁺ T cell population produced by the method of claim 21.

25. A method for stimulating a population of CD8⁺ T cells to proliferate, comprising:

a) obtaining peripheral blood leukocytes from an individual;

b) isolating a population of CD8⁺ T cells from the peripheral blood leukocytes by negative selection with a combination of antibodies directed to surface markers unique to the cells negatively selected;

c) contacting the population of CD8⁺ T cells with an anti-CD3 antibody immobilized on a solid phase and a ligand which binds a CD9 antigen present on activated T cells, under conditions appropriate for stimulating proliferation of the T cells;

d) separating the anti-CD3 antibody from the T cells and the ligand;

e) monitoring proliferation of the T cells in response to continuing exposure to the ligand by examining cell size; and

f) restimulating the T cells with the anti-CD3 antibody and the ligand when T cell size has decreased to induce further proliferation of the T cells.

26. The method of claim 25, further comprising repeating steps (d)-(f) to produce a population of CD8⁺ T cells increased in number of from about 100- to about 100,000-fold the original T cell population.

27. The method of claim 25, wherein the ligand binds a peptide comprising an amino acid sequence

(Xaa₁)_n -Gly-Xaa₂ -Trp-Leu-Xaa₃ -Xaa₄ -Asp(Glu)-(Xaa₅)_n (SEQ ID NO:

5),

wherein Xaa₄ may or may not be present, Xaa₁, Xaa₂, Xaa₃, Xaa₄ and Xaa₅ are any amino acid residue and n =0-20.

28. The method of claim 27, wherein Xaa₂ is Cys, Ile or Leu, Xaa₃ is Leu or Arg and Xaa₄, if present, is Arg, Pro or Phe.

29. The method of claim 25, wherein the population of CD8⁺ T cells are CD8⁺ T cells obtained from an individual afflicted with cancer and the method further comprises restoring the T cells to the individual.

30. A method for inducing a population of CD8⁺ T cells to proliferate, comprising:

a) activating a population of T cells; and

b) stimulating the population of T cells with a ligand which binds an antigen on the activated T cells, the antigen being recognized by monoclonal antibody ES5.2D8, the activating and stimulating steps thereby inducing proliferation of the T cells.

31. The method of claim 30, wherein the ligand binds an epitope recognized by monoclonal antibody ES5.2D8.

32. The method of claim 30, wherein the ligand binds a CD9 antigen.

33. The method of claim 30, wherein the CD8⁺ cells are preferentially expanded from a population of T cells.