GENERATING CIK NKT CELLS FROM CORD BLOOD
Provided herein are methods and customized media compositions for culturing CIK NKT cells.
1 . A population of CIK NKT cells, wherein greater than 50% of the cells in the population express CD 56 and CD3 and less than 10% of the cells in the population express Va24.
2 . The population of CIK NKT cells of claim 1 , wherein the CIK NKT cells can kill a target cell in the absence of alpha-galactosylceramide (Gal-Cer).
3 . The population of CIK NKT cells of claim 1 , wherein the target cell is a cancer cell.
4 . The population of CIK NKT cells of claim 1 , wherein the cancer cell line is selected from the group consisting of a myelogenous leukemia cell, a medulloblastoma cell, and a monocytic cell.
5 . The population of CIK NKT cells of claim 3 , wherein the cancer cell is selected from the group consisting of a K562 cell, a Daudi cell, a DAOY cell, and a THP-1 cell.
6 . The population of CIK NKT cells of claim 2 , wherein the CIK NKT cells kill a plurality of the target cells at an EC50 of between 1.0 and 10.0.
7 . The population of CIK NKT cells of claim 6 , wherein the CIK NKT cells can kill the target cells at a EC50 that is no less than 90% and no greater than 110% of the EC50 at which the CIK NKT cells killing the target cells in the presence of Gal-Cer.
8 . A composition comprising a plurality of CIK NKT cells from the population of CIK NKT cells of claim 1 , and a physiologically acceptable excipient.
9 . (canceled)
10 . A method of enriching CIK NKT cells from a cord blood sample, the method comprising:
isolating mononuclear cells from the cord blood sample; and
contacting the isolated mononuclear cells with one or more agents selected from the group consisting of IL-7, ALT-803 or IL-15, FLT3 ligand, and Gal-Cer, whereby enriching CIK NKT cells.
11 . The method of claim 10 , wherein the IL-7, if present, is in a concentration ranging from 5 to 20 ng/mL.
12 . The method of claim 10 , wherein the ALT-803, if present, is in a concentration ranging from 100 to 300 ng/mL.
13 . The method of claim 10 , wherein the FLT3 ligand, if present, is in a concentration ranging from 5 to 20 ng/mL.
14 . The method of claim 10 , wherein the Gal-Cer is present in a concentration ranging from 2 to 10 μg/mL.
15 . The method of claim 10 , wherein the method further comprises isolating the enriched CIK NKT cells from the rest of the cord blood sample.
16 . The method of claim 15 , wherein the method further comprises contacting the isolated CIK NKT cells with anti-CD3, anti-CD28, and IL2 to expand the CIK NKT cells.
17 . The method of claim 16 , wherein the method further comprises contacting the isolated CIK NKT cells with Gal-Cer.
18 . The method of claim 17 , wherein the Gal-Cer is a present in a form of a Gal-Cer loaded CD1d tetramer.
19 . The method of claim 16 , wherein the anti-CD3 antibody is present in an amount of 5 ng/mL to 60 ng/mL.
20 . The method of claim 16 , wherein the anti-CD28 antibody is present in an amount of 0.1 μg/mL to 2 μg/mL.
21 . The method of claim 16 , wherein IL-2 is present in a concentration of 50 ng/mL to 500 ng/mL.
22 . The method of claim 16 , wherein the production of CIK NKT cells does not include interferon-gamma.
23 . A method of treating cancer or viral infection in a patient in need thereof, the method comprising administering to the patient a therapeutically effective amount of CIK NKT cells from the population of CIK NKT cells of claim 1 , thereby treating cancer.
24 . The method of claim 23 , wherein about 1×10 8 to about 1×10 11 cells per m 2 of body surface area of the patient are administered to the patient.
25 . The method of claim 23 , wherein the cancer is selected from the group consisting of a leukemia, a lymphoma, polycythemia vera, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, a sarcoma and a carcinoma.
26 . The method of claim 25 , wherein the cells are administered to the patient by a route selected from the group consisting of intravenous, intraperitoneal, and subcutaneous.
27 . The method of claim 23 , wherein the method further comprises administering an antibody.
28 . A population of CIK NKT cells produced by the methods of claim 10 .
29 . The population of CIK NKT cells of claim 1 , wherein the CIK NKT cells express a CAR and/or a cytokine.