Methods of T cell expansion and activation
The present disclosure relates to methods, cells, and compositions for preparing T cell populations and compositions for adoptive cell therapy. In particular, provided herein are methods for efficiently expanding and activating T cell populations for genetic engineering and adoptive T cell immunotherapies. Also provided are cells and compositions produced by the methods and methods of their use.
1. A method of preparing a population of T cells, the method comprising
(a) reducing BAFF-R receptor activity in the T cells; and
(b) culturing the T cells of (a) for about 3 to about 14 days in the presence of an anti-CD3 antibody and an anti-CD28 antibody under conditions appropriate for activating cytotoxic T cells,
wherein the reducing and culturing activates and induces proliferation of activated T cells to yield a population comprising activated T cells in sufficient numbers for use in therapy.
2. The method of claim 1 , wherein the T cells are selected from the group consisting of a leukocyte-containing cell mixture and a purified T cell population.
3. The method of claim 2 , wherein the leukocyte-containing cell mixture or purified T cell population is obtained from apheresis of peripheral blood of a human subject.
4. The method of claim 2 , wherein the leukocyte-containing cell mixture or purified T cell population is obtained from peripheral blood mononuclear cells of a human subject.
5. The method of claim 1 , wherein the population comprises at least one of activated CD4 + T cells and CD8 + T cells.
6. The method of claim 1 , wherein cytotoxic CD8 + T cells are preferentially expanded from the activated T cell population.
7. The method of claim 1 , wherein the method of step (a) is selected from the group consisting of:
(i) culturing T cells in the presence of a BAFF-R antagonist; and
(ii) contacting T cells with a BAFF-R specific shRNA.
8. The method of claim 7 , wherein the BAFF-R antagonist is a neutralizing BAFF-R antibody.
9. A method of claim 1 additionally comprising the step of:
(c) providing in the T cells a chimeric antigen receptor, to generate a population of activated T cells comprising the chimeric antigen receptor.
10. The method of claim 9 , wherein the providing step (c) comprises a method step selected from the group consisting of:
(i) introducing the chimeric antigen receptor into the T cells; and
(ii) transfecting a nucleic acid vector encoding the chimeric antigen receptor into the T cells whereby the T cells express the chimeric antigen receptor.
11. The method of claim 9 , wherein the reducing step (a) comprises a method step selected from the group consisting of:
(i) culturing T cells in the presence of a BAFF-R antagonist; and
(ii) contacting T cells with a BAFF-R specific shRNA.
12. The method of claim 11 , wherein the BAFF-R antagonist is a neutralizing BAFF-R antibody.
13. A method of preparing a population of T cells, the method comprising
(a) reducing BAFF-R receptor activity in the T cells; and
(b) culturing the T cells of (a) for about 3 to about 14 days in the presence of an anti-CD3 antibody, or a CD3-binding fragment thereof, and an anti-CD28 antibody, or a CD28-binding fragment thereof, under conditions appropriate for activating cytotoxic T cells,
wherein the reducing and culturing activates and induces proliferation of activated T cells to yield a population comprising activated T cells in sufficient numbers for use in therapy.
14. The method of claim 13 , wherein the anti-CD3 antibody is an anti-CD3ε antibody.
15. The method of claim 14 , wherein the anti-CD3ε antibody is selected from the group consisting of clone UCHT1 and clone OKT3.
16. The method of claim 13 , wherein the anti-CD28 antibody is selected from the group consisting of clone 37407 and clone 9.3.
17. The method of claim 13 , wherein the anti-CD3 and anti-CD28 antibodies are attached to a substrate.
18. The method of claim 17 , wherein the substrate is a plate or a bead.
19. The method of claim 13 , wherein the T cells are selected from the group consisting of a leukocyte-containing cell mixture and a purified T cell population.
20. The method of claim 19 , wherein the leukocyte-containing cell mixture or purified T cell population is obtained from apheresis of peripheral blood of a human subject.
21. The method of claim 13 , wherein the population comprises at least one of activated CD4 + T cells and CD8 + T cells.
22. The method of claim 13 , wherein the method of step (a) is selected from the group consisting of:
(i) culturing T cells in the presence of a BAFF-R antagonist; and
(ii) contacting T cells with a BAFF-R specific shRNA.
23. The method of claim 22 , wherein the BAFF-R antagonist is a neutralizing BAFF-R antibody.
24. A method of claim 13 additionally comprising the step of:
(c) providing in the T cells a chimeric antigen receptor, to generate a population of activated T cells comprising the chimeric antigen receptor.
25. The method of claim 24 , wherein the providing step (c) comprises a method step selected from the group consisting of:
(i) introducing the chimeric antigen receptor into the T cells; and
(ii) transfecting a nucleic acid vector encoding the chimeric antigen receptor into the T cells whereby the T cells express the chimeric antigen receptor.
26. The method of claim 24 , wherein the reducing step (a) comprises a method step selected from the group consisting of:
(i) culturing T cells in the presence of a BAFF-R antagonist; and
(ii) contacting T cells with a BAFF-R specific shRNA.
27. The method of claim 26 , wherein the BAFF-R antagonist is a neutralizing BAFF-R antibody.