Adoptive transfer of CD8+ T cell clones derived from central memory cells
The present invention provides a method of carrying out adoptive immunotherapy in a primate subject in need thereof by administering the subject a cytotoxic T lymphocytes (CTL) preparation in a treatment-effective amount. The method comprises administering as the CTL preparation a preparation consisting essentially of an in vitro expanded primate CTL population, the CTL population enriched prior to expansion for central memory T lymphocytes, and depleted prior to expansion of effector memory T lymphocytes. In some embodiments, the method may further comprise concurrently administering Interleukin-15 to the subject in an amount effective to increase the proliferation of the central memory T cells in the subject. Pharmaceutical formulations produced by the method, and methods of using the same, are also described.
1. A method for performing adoptive immunotherapy, comprising administering to a primate subject in need thereof a treatment-effective amount of a cytotoxic T lymphocyte (CTL) preparation, wherein the CTL preparation comprises 60% or more primate CD8+ effector T (T E ) cells and one or more of the CD8+ T E cells of the CTL preparation comprise a gene encoding a protein that specifically binds to a cancer cell, and wherein the 60% or more primate CD8+ T E cells have:
(a) decreased expression of CD62L as compared to CD8 + central memory T (T CM ) cells; and
(b) improved survival and/or expansion for at least 10-days when cultured in vitro in the presence of IL-15 and in the absence of antigen stimulation as compared to when cultured in vitro in the absence of IL-15 and in the absence of antigen stimulation.
2. The method according to claim 1 , wherein the CD8+ T E cells of the CTL preparation are positive for granzyme B, perforin or both.
3. The method according to claim 1 , wherein the CTL preparation comprises 70%, 80%, 85% or 90% or more of the primate CD8+ T E cells having the decreased expression of CD62L.
4. The method according to claim 1 , wherein the CTL preparation consists essentially of the primate CD8+ T E cells having the decreased expression of CD62L.
5. The method according to claim 1 , wherein the CD8+ T E cells of the CTL preparation have decreased expression of CCR7, CD28, and CD127 as compared to CD8+ T CM cells.
6. The method according to claim 1 , wherein the 60% or more of CD8+ T E cells of the CTL preparation have improved survival and/or expansion in vitro for at least 34 days in the presence of IL-15.
7. The method according to claim 1 , wherein the gene encoding a protein that specifically binds to the target on the cell is an engineered immunoreceptor.
8. The method according to claim 1 , wherein the CTL preparation further comprises IL 15.
9. The method according to claim 1 , wherein the CTL preparation is formulated in a pharmaceutically acceptable carrier.
10. The method according to claim 1 , wherein the CD8+ T E cells of the CTL preparation have increased expression of CD62L after adoptive immunotherapy as compared to the CD8+ T E cells of the CTL preparation before adoptive immunotherapy.
11. The method according to claim 1 , wherein the CD8+ T E cells of the CTL preparation migrate to memory T cell niches in vivo.
12. The method according to claim 11 , wherein the in vivo memory T cell niches comprise lymph nodes, bone marrow, or both.
13. The method according to claim 1 , wherein the CD8+ T E cells of the CTL preparation persist in vivo upon adoptive transfer for at least 5 days, at least 7 days, at least 10 days, at least 14 days, at least 21 days, at least 28 days, at least 42 days, at least 56 days, at least 70 days, at least 77 days, at least 215 days, or at least 344 days.
14. The method according to claim 1 , wherein the subject has cancer.
15. The method according to claim 14 , wherein the cancer is selected from colon cancer, lung cancer, liver cancer, breast cancer, prostate cancer, ovarian cancer, skin cancer, bone cancer, brain cancer, breast cancer, squamous cell carcinoma, melanoma, leukemia, or myeloma.
16. The method according to claim 1 , wherein the cancer cell is CD19+ or CD20+.
17. The method according to claim 1 , wherein the CTL preparation is allogeneic to the subject, syngeneic to the subject, or autologous to the subject.
18. The method according to claim 1 , wherein the primate subject is a human.
19. A method for performing adoptive immunotherapy of a primate subject having a cytomegalovirus (CMV) or Epstein-Barr virus (EBV) infection, comprising administering to the primate subject in need thereof a treatment-effective amount of a cytotoxic T lymphocyte (CTL) preparation, wherein the CTL preparation comprises 60% or more primate CD8+ effector T (T E ) cells and one or more of the CD8+ T E cells of the CTL preparation comprise a gene encoding a T-cell receptor (TCR) that specifically binds to a CMV or EBV infected cell, and wherein the 60% or more primate CD8+ T E cells have:
(a) decreased expression of CD62L as compared to CD8 + central memory T (T CM ) cells; and
(b) improved survival and/or expansion for at least 10 days when cultured in vitro in the presence of IL-15 and in the absence of antigen stimulation as compared to when cultured in vitro in the absence of IL-15 and in the absence of antigen stimulation.
20. The method according to claim 19 , wherein the CTL preparation comprises 70%, 80%, 85% or 90% or more of the primate CD8+ T E cells having the decreased expression of CD62L.
21. The method according to claim 19 , wherein the CTL preparation further comprises IL 15.