Antigen presenting T cells, sensitized, manufactured T cells and methods of treatment using the same
The present disclosure provides methods for treating cancer and infectious disease using manufactured T cells or other cell types including antigen-presenting T cells, loaded, antigen-presenting T cells, sensitized, manufactured T cells and combinations thereof which can be co-administered with the manufactured T cells or without. Methods of treatment can include administration of pentostatin and cyclophosphamide followed by administration of T cells. The present disclosure also provides methods to produce antigen-presenting T cells from the manufactured T cells, load antigen-presenting T cells and to sensitize the manufactured T cells using the loaded, antigen-presenting T cells. Methods of treatment using the antigen-presenting T cells and/or sensitized, manufactured T cells are also provided which can be performed in vivo or ex vivo.
1 . A method for producing manufactured T cells, comprising:
inoculating a culture input population of cells from a subject at a cell density of at least 1.5×10 6 cells per mL in a culture medium comprising temsirolimus and an IL-2 signaling inhibitor, wherein said temsirolimus is present in said culture medium at a concentration of at least 1 μM, wherein the culture input population of cells comprises T cells;
incubating said culture input population of cells in the culture medium for a first period of time, wherein said first period of time is at least 16 hours;
adding IFN-α, IL-7, and IL-15 to the culture medium after the first period of time, wherein said IFN-α is added to said culture medium to a concentration of at least 10,000 IU/mL;
incubating said culture input population of cells in the culture medium for a second period of time following addition of IFN-α, IL-7, and IL-15 to yield manufactured T cells, wherein IL-2 is not added to the culture medium at any time during the method, and wherein no anti-CD3/anti-CD28 co-stimulation is performed at any time during the method; and
harvesting said manufactured T cells, wherein 5% or less of the manufactured T cells that are CD4+ or CD8+ express CD25 as measured by flow cytometry.
2 . The method of claim 1 , further comprising, after harvesting said manufactured T cells:
packaging at least a portion of said manufactured T cells in a package; and
freezing said package containing said portion of said manufactured T cells.
3 . The method of claim 1 , wherein said IL-2 signaling inhibitor is an anti-IL-2 receptor antibody or fragment thereof.
4 . The method of claim 3 , wherein said anti-IL-2 receptor antibody is basiliximab or daclizumab.
5 . The method of claim 1 , wherein said IL-2 signaling inhibitor is present in said culture medium at a concentration of 5 to 50 μg/mL.
6 . The method of claim 1 , wherein said first and second period of time is a total of 6 days.
7 . The method of claim 1 , wherein the culture medium is free of serum.
8 . The method of claim 1 , wherein said culture input population of cells comprises about 99% or more T cells out of the total number of cells in said culture input population of cells.
9 . A method for treating a solid tumor in a subject comprising:
obtaining a sample of immune cells harvested from the subject, wherein the sample of immune cells comprises T cells;
adding the sample of immune cells to a culture medium at a cell density of at least 1.5×10 6 cells per mL, wherein the culture medium comprises temsirolimus and an IL-2 signaling inhibitor, wherein said temsirolimus is present in said culture medium at a concentration of at least 1 UM;
incubating said sample of immune cells in the culture medium for a first period of time, wherein said first period of time is at least 16 hours;
adding IFN-α, IL-7, and IL-15 to the culture medium after the first period of time, wherein said IFN-α is added to said culture medium to a concentration of at least 10,000 IU/mL;
incubating said sample of immune cells in the culture medium for a second period of time following addition of IFN-α, IL-7, and IL-15 to yield manufactured T cells, wherein IL-2 is not added to the culture medium at any time during the production of manufactured T cells, and wherein no anti-CD3/anti-CD28 co-stimulation is performed at any time during the method production of manufactured T cells; and
harvesting said manufactured T cells, wherein 5% or less of the manufactured T cells that are CD4+ or CD8+ express CD25 as measured by flow cytometry; and
administering a therapeutically effective amount of the manufactured T cells to the subject.
10 . The method of claim 9 , wherein the method further comprises administering to the subject a chemotherapeutic agent in an amount sufficient to induce immunogenic cell death of at least a portion of cancer cells in the solid tumor.
11 . The method of claim 10 , wherein the chemotherapeutic agent sufficient to induce immunogenic cell death is selected from the group consisting of etoposide, bortezomib, doxorubicin, epirubicin, cyclophosphamide, oxaliplatin, 5-fluorouracil, gemcitabine, mitoxantrone, bleomycin, dactinomycin, lubrinectedin, teniposide, radiation, cryotherapy, and combinations thereof.
12 . The method of claim 9 , wherein the therapeutically effective amount of manufactured T cells is about 0.1×10 6 cells/kg to about 5×10 6 cells/kg of subject body weight.
13 . The method of claim 9 , wherein the solid tumor is selected from the group consisting of renal cell carcinoma, bladder cancer, lung cancer, small-cell lung cancer, non-small cell lung cancer, liver cancer, gastric cancer, colon cancer, sarcoma, pancreatic cancer, prostate cancer, ovarian cancer, breast cancer, rectal cancer, endometrial cancer, kidney cancer, bile duct cancer, thyroid cancer, melanoma, non-melanoma skin cancer, esophageal cancer, cervical cancer, oral cavity cancer, pharyngeal cancer, stomach cancer, brain cancer, and colorectal cancer.