Methods for selectively stimulating proliferation of T cells
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.
1. A method for expanding a population of CD8 + T cells in number of from about 100 to about 100,000 fold over the original CD8 + T cell population or to about 10 log 10 to 12 log 10 , comprising;
a) contacting a population of CD8 + T cells with a surface comprising
1) an anti-CD3 antibody or a CD-3-binding fragment thereof; and
2) a second agent which stimulates a CD28 antigen, wherein the second agent is selected from the group consisting of an anti-CD28 antibody or a CD28-binding fragment thereof, B7-1 or a CD28-binding fragment thereof, B7-2 or a CD28-binding fragment thereof, a B7-1Ig fusion protein that interacts with CD28 and a B7-2Ig fusion protein that interacts with CD28, wherein said anti-CD3 antibody or CD3-binding fragment thereof, and said second agent are attached on the same surface;
b) adding exogenous IL-2 to the population of CD8 + T cells, such that the population of CD8 + T cells increase in number from about 100 to about 100,000 fold over the original CD8 + T cell population or to about 10 log 10 to 12 log 10 .
2. The method of claim 1 , wherein the anti-CD3 antibody or the CD3-binding fragment thereof is a monoclonal antibody or a CD3-binding fragment thereof.
3. The method of claim 2 , wherein the anti-CD3 monoclonal antibody or the CD3-binding fragment thereof is OKT 3- or a CD3-binding fragment thereof.
4. The method of claim 1 , wherein the second agent is an anti CD-28 monoclonal antibody or a CD28-binding fragment thereof.
5. The method of claim 4 , wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is a human anti-CD28 antibody or a CD28-binding fragment thereof.
6. The method of claim 1 , wherein IL-2 is added to the population of T cells in an amount sufficient to maintain proliferation of the T cells.
7. The method of claim 1 , 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.
8. The method for preparing a renewable source of CD8 + T cells, comprising expanding a population of CD8 + 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.
9. The method of claim 1 , wherein said surface is a bead.
10. The method of claim 9 , wherein the bead is a magnetic immunobead.
11. The method of claim 9 , wherein the bead is a polystyrene bead.
12. The method of claim 1 , wherein said surface is a cell surface.
13. The method of claim 1 , wherein said surface is a tissue culture dish.
14. The method of claim 1 , wherein the anti-CD3 antibody or CD3-binding fragment thereof, and the second agent are covalently attached on the same surface.
15. The method of claim 1 , wherein the anti-CD3 antibody or CD3-binding fragment thereof, and the second agent are attached on the same surface via an avidin-biotin complex.
16. The method of claim 1 , wherein the anti-CD3 antibody or CD3-binding fragment thereof, and the second agent are directly attached on the same surface.
17. The method of claim 4 , wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is EX5.3D10 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB311373.
18. The method of claim 4 , wherein the anti-CD28 monoclonal antibody or the CD28-binding fragment thereof is 9.3 or a CD28-binding fragment thereof obtainable from a hybridoma deposited as ATCC Deposit No. HB10271.
19. The method of claim 1 , wherein the second agent is a B7-1Ig fusion protein that interacts with CD28.
20. The method of claim 19 , wherein the B7-1Ig fusion protein comprises about amino acids 1–208 of SEQ ID NO:2.
21. The method of claim 19 , wherein the Ig component of the B7-1Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.
22. The method of claim 1 , wherein the second agent is a B7-2Ig fusion protein that interacts with CD28.
23. The method of claim 22 , wherein the B7-2Ig fusion protein comprises about amino acids 24–245 of SEQ ID NO:4.
24. The method of claim 22 , wherein the Ig component of the B7-2Ig fusion protein is selected from the group consisting of an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG1, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG2, an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG3 and an Fc fragment that comprises the hinge, CH2 and CH3 regions of human IgG4.