Processes for production of tumor infiltrating lymphocytes and uses of same in immunotherapy
The present invention provides improved and/or shortened methods for expanding TILs and producing therapeutic populations of TILs, including novel methods for expanding TIL populations in a closed system that lead to improved efficacy, improved phenotype, and increased metabolic health of the TILs in a shorter time period, while allowing for reduced microbial contamination as well as decreased costs. Such TILs find use in therapeutic treatment regimens.
1. A method for treating a subject with a cancer, the method comprising administering expanded tumor infiltrating lymphocytes (TILs) comprising:
(a) adding dissociated tumor materials into a closed system, wherein the dissociated tumor materials comprise a first population of TILs and are obtained from a tumor that was resected from the subject;
(b) performing a first expansion by culturing the first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is performed in a closed container providing a first gas-permeable surface area, wherein the first expansion is performed for a first period of about 3 to 11 days to obtain the second population of TILs, and wherein the transition from step (a) to step (b) occurs without opening the system;
(c) performing a second expansion by supplementing the cell culture medium with additional IL-2, OKT-3, and antigen presenting cells (APCs) to produce a third population of TILs, wherein the second expansion is performed for a second period of about 7 to 11 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs, wherein the second expansion is performed in a closed container providing a second gas-permeable surface area, and wherein the transition from step (b) to step (c) occurs without opening the system;
(d) harvesting the third population of TILs obtained from step (c), wherein the transition from step (c) to step (d) occurs without opening the system;
(e) transferring the harvested third TIL population from step (d) to an infusion bag, wherein the transfer from step (d) to (e) occurs without opening the system;
(f) cryopreserving the infusion bag comprising the harvested TIL population from step (e) using a cryopreservation process; and
(g) administering a therapeutically effective dosage of the harvested TIL population from the infusion bag in step (f) to the subject.
2. The method according to claim 1 , wherein the dissociated tumor materials comprise a tumor digest.
3. The method according to claim 1 , wherein the dissociated tumor materials comprise one or more tumor fragments.
4. The method according to claim 1 , wherein obtaining the dissociated tumor materials comprises mechanically disrupting the tumor resected from the subject.
5. The method according to claim 1 , wherein obtaining the dissociated tumor materials comprises enzymatically disrupting the tumor resected from the subject.
6. The method according to claim 1 , wherein the dissociated tumor materials have been previously cryopreserved and are thawed prior to step (a).
7. The method according to claim 1 , wherein the third population of TILs harvested in step (d) comprises sufficient TILs for administering a therapeutically effective dosage of the TILs in step (g).
8. The method according to claim 7 , wherein the therapeutically effective dosage in step (g) comprises from about 1×10 9 to about 9×10 10 TILs.
9. The method according to claim 1 , wherein the APCs comprise peripheral blood mononuclear cells (PBMCs).
10. The method according to claim 9 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.
11. The method according to claim 1 , wherein prior to administering a therapeutically effective dosage of TIL cells in step (g), a non-myeloablative lymphodepletion regimen has been administered to the subject.
12. The method according to claim 11 , where the non-myeloablative lymphodepletion regimen comprises the steps of administration of cyclophosphamide at a dose of 60 mg/m 2 /day for two days followed by administration of fludarabine at a dose of 25 mg/m 2 /day for five days.
13. The method according to claim 1 , further comprising the step of treating the subject with a high-dose IL-2 regimen starting on the day after administration of the TIL cells to the subject in step (g).
14. The method according to claim 13 , wherein the high-dose IL-2 regimen comprises 600,000 or 720,000 IU/kg administered as a 15-minute bolus intravenous infusion every eight hours until tolerance.
15. The method according to claim 1 , wherein the cancer is selected from the group consisting of melanoma (including metastatic melanoma), ovarian cancer, cervical cancer, non-small-cell lung cancer (NSCLC), lung cancer, bladder cancer, breast cancer, cancer caused by human papilloma virus, head and neck cancer (including head and neck squamous cell carcinoma (HNSCC)), renal cancer, and renal cell carcinoma.
16. The method according to claim 1 , wherein the cancer is melanoma (including metastatic melanoma).
17. The method according to claim 1 , wherein the first expansion in step (b) and the second expansion in step (c) are each individually performed within a period of 11 days.
18. The method according to claim 1 , wherein steps (a) through (e) are performed in about 10 days to about 22 days.
19. The method according to claim 1 , wherein the closed container in step (b) and/or step (c) is a gas-permeable bag.