Method and compositions for cellular immunotherapy
The present invention provides methods and compositions to confer and/or augment immune responses mediated by cellular immunotherapy, such as by adoptively transferring genetically modified tumor specific CD8+ T cells in the presence of tumor-specific, subset specific genetically modified CD4+ T cells, wherein the CD4+ T cells confer and/or augment a CD8+ T cells ability to sustain anti-tumor reactivity and increase and/or maximize tumor-specific proliferation of the tumor-specific CD8+ T cells of interest. Pharmaceutical formulations produced by the method, and methods of using the same, are also described.
1. An adoptive cellular immunotherapy composition containing chimeric antigen receptor-modified CD4 + T lymphocytes and chimeric antigen receptor-modified CD8 + T lymphocytes, wherein:
(a) the chimeric antigen receptor-modified CD4+ T lymphocytes contain a chimeric antigen receptor that specifically binds to an antigen and at least 50% of the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition are surface positive for CD62L and/or CD45RA, and
(b) the chimeric antigen receptor-modified CD8+ T lymphocytes contain a chimeric antigen receptor that specifically binds to the antigen and at least 50% of CD8+ cytotoxic T lymphocytes in the composition are surface positive for CD62L and/or CD45RO.
2. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is associated with a disease or disorder selected from a solid tumor, hematologic malignancy, melanoma, and infection with a pathogen.
3. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is selected from ROR1, tEGFR, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, and CEA.
4. The adoptive cellular immunotherapy composition according to claim 1 , wherein the chimeric antigen receptor of (a) and/or (b) comprises an extracellular antibody variable domain or single-chain antibody fragment specific for the antigen and an intracellular signaling module.
5. The adoptive cellular immunotherapy composition according to claim 4 , wherein each of the intracellular signaling module of the chimeric antigen receptor contained by the CD4+ T lymphocytes and the intracellular signaling module of the chimeric antigen receptor contained by the CD8+ T lymphocytes, individually, comprise (a) a CD28 costimulatory domain and a CD3 intracellular signaling domain, or (b) a 4-1BB costimulatory domain and a CD3 intracellular signaling domain.
6. The adoptive cellular immunotherapy composition according to claim 1 , wherein:
(a) at least 60% of the chimeric antigen receptor-modified CD4 + T lymphocytes are surface positive for CD62L and/or CD45RA;
(b) at least 60% of the chimeric antigen receptor-modified CD8 + cytotoxic T lymphocytes are surface positive for CD62L and/or CD45RO;
(c) at least 80% of the chimeric antigen receptor-modified CD4+ T lymphocytes are surface positive for CD62L and/or CD45RA; and/or
(d) at least 80% of the chimeric antigen receptor-modified CD8 + cytotoxic T lymphocytes are surface positive for CD62L and/or CD45RO.
7. The adoptive cellular immunotherapy composition according to claim 1 , wherein the intracellular signaling domain of the chimeric antigen receptor contained by the CD8+ T lymphocytes is the same as the intracellular signaling domain of the chimeric antigen receptor contained by the CD4+ T lymphocytes.
8. The adoptive cellular immunotherapy composition according to claim 1 , wherein the CD4+ helper T lymphocytes and the CD8+ cytotoxic T lymphocytes are derived from a subject and wherein the CD4 + helper T lymphocytes secrete a greater amount of IL-2 and/or exhibit a greater degree of proliferation when cultured in the presence of the antigen as compared to a reference population of CD4 + T cells that has been expanded in vitro from an effector memory T cell-enriched CD4 + T cell population or a bulk CD4 + T cell population from the subject and modified to contain the chimeric antigen receptor contained by the CD4+ helper T lymphocytes.
9. The adoptive cellular immunotherapy composition of claim 1 , wherein at least 50% of the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition are surface positive for CD62L.
10. The adoptive cellular immunotherapy composition of claim 1 , wherein at least 50% of the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes in the composition are surface positive for CD62L.
11. The adoptive cellular immunotherapy composition of claim 1 , wherein at least 50% of the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition are surface positive for CD62L and at least 50% of the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes in the composition are surface positive for CD62L.
12. The adoptive cellular immunotherapy composition of claim 1 , wherein at least 60% of the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition are surface positive for CD62L and/or at least 60% of the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes in the composition are surface positive for CD62L.
13. The adoptive cellular immunotherapy composition of claim 1 , wherein at least 80% of the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition are surface positive for CD62L and/or at least 80% of the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes in the composition are surface positive for CD62L.
14. The adoptive cellular immunotherapy composition of claim 1 , wherein the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition and the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes are present in the composition at a ratio of between about 2:1 to about 1:2.
15. The adoptive cellular immunotherapy composition of claim 1 , wherein the chimeric antigen receptor-modified CD4+ helper T lymphocytes in the composition and the chimeric antigen receptor-modified CD8+ cytotoxic T lymphocytes are present in the composition at a ratio of about 2:1, 1:1, or 1:2.
16. The adoptive cellular immunotherapy composition of claim 1 , wherein the chimeric antigen receptor contained by the CD8+ T lymphocytes is the same as the chimeric antigen receptor contained by the CD4+ T lymphocytes.
17. The adoptive cellular immunotherapy composition of claim 1 , wherein the intracellular signaling domain of the chimeric antigen receptor in the CD8+ T lymphocytes is different from the intracellular signaling domain of the chimeric antigen receptor in the CD4+ T lymphocytes; or the chimeric antigen receptor contained by the CD8+ T lymphocytes is different than the chimeric antigen receptor contained by the CD4+ T lymphocytes.
18. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is ROR1.
19. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is Her2.
20. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is L1-CAM.
21. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is CD19.
22. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is CD20.
23. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is CD22.
24. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is mesothelin.
25. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is CEA.
26. The adoptive cellular immunotherapy composition according to claim 2 , wherein the disease or disorder is a solid tumor, hematologic malignancy, or melanoma.
27. The adoptive cellular immunotherapy composition according to claim 2 , wherein the disease or disorder is a hematologic malignancy.
28. The adoptive cellular immunotherapy composition according to claim 1 , wherein the antigen is a tumor associated antigen associated with a cancer selected from the group consisting of melanoma, breast cancer, squamous cell carcinoma, colon cancer, leukemia, myeloma, prostate cancer, B cell lymphoma, breast cancer, prostate cancer, and leukemia.
29. The adoptive cellular immunotherapy composition according to claim 1 , wherein a greater percentage of the chimeric antigen receptor-modified CD4 + helper T lymphocytes are surface positive for CD62L and/or CD45RA as compared to a reference population of CD4 + T cells that has been expanded in vitro from an effector memory-enriched CD4 + T cell population or a bulk CD4 + T cell population from the subject and modified to contain the chimeric antigen receptor contained by the CD4+ helper T lymphocytes.
30. The adoptive cellular immunotherapy composition according to claim 1 , wherein a greater percentage of the chimeric antigen receptor-modified CD8 + cytotoxic T lymphocytes are surface positive for CD62L, CD45RO and/or CCR7 as compared to a reference population of CD8 + T cells that has been expanded in vitro from an effector memory-enriched CD8 + T cell population or bulk CD8 + T cell population from the subject and modified to contain the chimeric antigen receptor contained by the CD8+ cytotoxic T lymphocytes.
31. The adoptive cellular immunotherapy composition according to claim 1 , wherein the chimeric antigen receptor-modified CD8 + cytotoxic T lymphocytes proliferate when co-cultured in vitro with the CD4 + helper T lymphocytes in the presence of the antigen, to a greater degree as compared to the degree of proliferation of the CD8 + cytotoxic T lymphocytes when co-cultured, under the same conditions, with a reference population of CD4 + T cells expanded in vitro from an effector memory-enriched CD4 + T cell population or a bulk CD4 + T cell population from the subject and modified to contain the chimeric antigen receptor contained by the CD4+ helper T lymphocytes.