Methods for selectively stimulating proliferation of T cells

• US 6,905,681 B1
• Filed: 07/08/1999
• Issued: 06/14/2005
• Est. Priority Date: 06/03/1994
• Status: Expired due to Fees

• Current Assignee: 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: 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)
• Inventors: Nabel, Gary J., June, Carl H., Thompson, Craig B., Gray, Gary S., Rennert, Paul D.
• Primary Examiner(s): GAMBEL, PHILLIP
• Application Number:
  US09/349,915
• Time in Patent Office: 2,168 Days
• Field of Search: 435/2, 435/7.1, 435/375, 530/387.1, 530/388.22, 530/388.75, 424/130.1
• US Class Current: 424/93.71
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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 39/464411 Immunoglobulin superfamily
• 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 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• 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 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/74 : Inducing cell proliferation
• 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 2795/00011 : Details
• 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 including ex vivo proliferation of a population of T cells to sufficient numbers for use in therapy, comprising contacting a population of T cells ex vivo with a surface having directly attached thereon:

(a) an anti-CD3 antibody or CD3-binding fragment thereof, which provides a primary activation signal to the T cells, thereby activating the T cells; and

(b) an anti-CD28 antibody or CD28-binding fragment thereof, which stimulates a CD28 accessory molecule on the surface of the T cells, thereby stimulating the activated T cells,

wherein the anti-CD3 antibody or CD3-binding fragment thereof, and the anti-CD28 antibody or CD28-binding fragment thereof, are directly attached on the same surface,

the anti-CD3 antibody or CD3-binding fragment thereof, and the anti-CD28 antibody or CD28-binding fragment thereof, thereby inducing the T cells to proliferate to sufficient numbers for use in therapy.

2. The method of claim 1, wherein the anti-CD3 antibody or CD3-binding fragment thereof is an anti-human CD3 monoclonal antibody or CD3-binding fragment thereof.

3. The method of claim 1, wherein the anti-CD28 antibody or CD28-binding fragment thereof is an anti-human CD28 monoclonal antibody or CD28-binding fragment thereof.

4. The method of claim 1, further comprising:

monitoring the proliferation of the T cells; and

reactivating and re-stimulating the T cells with the anti-CD3 antibody or CD3-binding fragment thereof, and the anti-CD28 antibody or CD28-binding fragment thereof, when the rate of T cell proliferation has decreased to induce further proliferation of the T cells.

5. The method of claim 4, wherein the step of monitoring proliferation of the T cells is by examining cell size or determining the level of expression of a cell surface molecule selected from the group consisting of B7-1, B7-2, and any combination thereof, and the step of reactivating and restimulating is initiated when T cell has decreased or when the level of the cell surface molecule has decreased.

6. The method of claim 1, wherein the T cells are induced to proliferate to about 100-fold the original T cell population.

7. The method of claim 1, wherein the T cells are induced to proliferate to about 100,000-fold the original T cell population.

8. The method of claim 1, wherein the T cells are induced to proliferate for at least 3 days.

9. The method of claim 1, wherein the T cells are induced to proliferate for at least 7 days.

10. The method of claim 1, wherein the surface is a bead.

11. The method of claim 10, wherein the bead is a magnetic bead.

12. The method of claim 10, wherein the bead is a polystyrene bead.

13. The method of claim 1, wherein the surface is a cell surface.

14. The method of claim 1, wherein the surface is a tissue culture dish.

15. The method of claim 1, wherein the population of T cells are induced to proliferate to sufficient numbers for use in treating cancer.

16. The method of claim 1, wherein the population of T cells are induced to proliferate to sufficient numbers for use in treating an infectious disease.

17. The anti-CD28 antibody of claim 3, wherein the antibody is 9.3, produced by the hybridoma assigned ATCC No. HB-10271.

18. The anti-CD28 antibody of claim 3, wherein the antibody is EX 5.3D10, produced by the hybridoma assigned ATCC No. HB-11373.