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 population of expanded TILs, wherein the population of expanded TILs is a therapeutic population of TILs obtainable by a method 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 a 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; and
(d) harvesting the therapeutic population of TILs obtained from step (c), wherein the transition from step (c) to step (d) occurs without opening the system.
2. The population of expanded TILs according to claim 1 , wherein the method further comprises the step (e) of transferring the harvested TIL population from step (d) to an infusion bag, wherein the transfer from step (d) to (e) occurs without opening the system.
3. The population of expanded TILs according to claim 2 , wherein the method optionally comprises the step (f) of cryopreserving the infusion bag comprising the harvested TIL population from step (e) using a cryopreservation process.
4. The population of expanded TILs according to claim 1 , wherein the dissociated tumor materials comprise a tumor digest.
5. The population of expanded TILs according to claim 1 , wherein the dissociated tumor materials comprise one or more tumor fragments.
6. The population of expanded TILs according to claim 1 , wherein obtaining the dissociated tumor materials comprises mechanically disrupting the tumor resected from the subject.
7. The population of expanded TILs according to claim 1 , wherein obtaining the dissociated tumor materials comprises enzymatically disrupting the tumor resected from the subject.
8. The population of expanded TILs according to claim 1 , wherein the dissociated tumor materials have been previously cryopreserved and are thawed prior to step (a).
9. The population of expanded TILs according to claim 1 , wherein the cell culture medium is CTS Optimizer.
10. The population of expanded TILs according to claim 1 , wherein the medium in the first expansion and/or the second expansion is free of human serum.
11. The population of expanded TILs according to claim 1 , wherein the therapeutic population of TILs harvested in step (d) comprises sufficient TILs for a therapeutically effective dosage of the TILs.
12. The population of expanded TILs according to claim 11 , wherein the number of TILs sufficient for a therapeutically effective dosage is from about 1×10 9 to about 9×10 10 .
13. The population of expanded TILs according to claim 1 , wherein the APCs are peripheral blood mononuclear cells (PBMCs).
14. The population of expanded TILs according to claim 13 , wherein the PBMCs are supplemented at a ratio of about 1:25 TIL:PBMCs.
15. The population of expanded TILs according to claim 1 , wherein the therapeutic population of TILs harvested in step (d) exhibits an increased subpopulation of CD8+ cells relative to the first and/or second population of TILs.
16. The population of expanded TILs 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.
17. The population of expanded TILs according to claim 1 , wherein steps (a) through (d) are performed in about 10 days to about 22 days.
18. The population of expanded TILs according to claim 1 , wherein steps (a) through (d) are performed in about 15 days to about 22 days.
19. The population of expanded TILs according to claim 1 , wherein steps (a) through (d) are performed in about 20 days to about 22 days.