Methods for treating HIV infected subjects

• US 20060099177A1
• Filed: 01/05/2006
• Published: 05/11/2006
• Est. Priority Date: 11/23/1988
• Status: Active Grant

• 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.
• Granted Patent: US 7,479,269 B2
• Time in Patent Office: Days
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• 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 16/2818 against CD28 or CD152
• 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 2317/55 Fab or Fab'
• 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 2740/16011 Human Immunodeficiency Virus, HIV

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 selectively enriching a population of T cells in TH1 cells in vitro comprising:

a) contacting a population of T cells with;

(1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and

(2) an anti-CD28 antibody or a CD28-binding fragment thereof, thereby stimulating the activated T cells, such that the T cells are stimulated to proliferate and produce at least one TH1 cytokine,

thereby selectively enriching the population of T cells in TH1 cells.

2. The method of claim 1, further comprising:

b) determining the amount of at least one TH1 cytokine, wherein an increase in the amount of the at least one TH1 cytokine following activation and stimulation of the T cells compared to the amount of the at least one TH1 cytokine produced by T cells that have not been activated as in (1) and stimulated in (2), indicates enrichment of the population of T cells in TH1 cells.

3. The method of claim 1, further comprising:

c) monitoring proliferation of the T cells; 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, such that the population of T cells is enriched in TH1 cells.

4. The method of claim 1, wherein the agent which provides a primary activation signal and the anti-CD28 antibody or CD28-binding fragment thereof are attached to a surface.

5. The method of claim 1, wherein the agent which provides a primary activation signal and the anti-CD28 antibody or CD28-binding fragment thereof are attached to the same surface.

6. The method of claim 5, wherein the surface is surface is a bead, a tissue culture flask, or a cell surface.

7. The method of claim 6, wherein the bead is a magnetic bead or a biodegradable bead.

8. The method of claim 3, wherein 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 combinations thereof, and the method further comprises reactivating and restimulating the T cells when T cell size has decreased or the level of the cell surface molecule has decreased to induce further proliferation of the T cells.

9. A method for selectively enriching a population of T cells in TH2 cells in vitro comprising

a) contacting a population of T cells with:

(1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and

(2) a stimulatory form of a natural ligand of CD28 selected from the group consisting of B7-1, a CD28-binding fragment of B7-1, B7-2, and a CD28-binding fragment of B7-2, such that the T cells are stimulated to proliferate and produce at least one TH2 cytokine,

thereby enriching the population of T cells in TH2 cells.

10. The method of claim 9, further comprising:

b) determining the amount of at least one TH2 cytokine, wherein an increase of the at least one TH2 cytokine following the activation and stimulation of the T cells compared to the amount of the at least one TH2 cytokine produced by T cells that have not been activated as in (1) and stimulated in (2), indicates enrichment of the population of T cells in TH2 cells.

11. The method of claim 9, further comprising:

c) monitoring the proliferation of the T cells; 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, such that the population of T cells is enriched in TH2 cells.

12. The method of claim 9, wherein the agent which provides a primary activation signal and the stimulatory form of a natural ligand of CD28 are attached to a surface.

13. The method of claim 9, wherein the agent which provides a primary activation signal and the stimulatory form of a natural ligand of CD28 are attached to the same surface.

14. The method of claim 13, wherein the surface is a bead, a tissue culture flask, or a cell surface.

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

16. The method of claim 11, wherein 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 combinations thereof, and the method further comprises reactivating and restimulating the T cells when T cell size has decreased or the level of the cell surface molecule has decreased to induce further proliferation of the T cells.

17. A method for increasing the proportion of TH1 versus TH2 cells in a population of T cells, comprising:

a) contacting the population of T cells with;

(1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and

(2) an anti-CD28 antibody or a CD28-binding fragment thereof, wherein the anti-CD28 antibody or a CD28-binding fragment is attached on a surface,

thereby increasing the proportion of TH1 versus TH2 cells in the population of T cells compared to the proportion of TH1 versus TH2 cells in a population of T cells not contacted with an agent which provides a primary activation signal to the T cells and an anti-CD28 antibody or a CD28-binding fragment thereof.

18. The method of claim 17, wherein the agent which provides a primary activation signal is attached to a surface.

19. The method of claim 18, wherein the agent which provides a primary activation signal and the anti-CD28 antibody or a CD28-binding fragment thereof are attached to the same surface.

20. The method of claim 19, wherein the surface is a bead, a tissue culture flask, or a cell surface.