Method of enhancing persistence of adoptively infused t cells

Abstract

The present disclosure provides for methods of improving the efficacy of T cells. In an aspect, the disclosure further provides for methods of enhancing the persistence of T cells for adoptive cell transfer or therapy (ACT). Cytokine sensitivity assays (CSA) and associated methodology capable of predicting the persistence of adoptively infused T Cells are further provided for by way of the instant disclosure. The disclosure also provides for methods of treating cancer in a subject in need thereof as well as T cells populations produced by methods described herein.

What is claimed is: 
     
         1 . A method of producing T cells comprising
 obtaining T cells from at least one individual,   activating the T cells,   expanding a first portion of the activated T cells over a period of time,   culturing the expanded T cells in the presence of at least one cytokine,   measuring a cytokine response in the cultured T cells,   identifying the period of time that yields a maximum cytokine response, and   expanding a second portion of the activated T cells for the period of time that yields a maximum cytokine response.   
     
     
         2 . The method of  claim 1 , further comprising freezing the expanded first portion of the activated T cells prior to culturing. 
     
     
         3 . The method of  claim 2 , further comprising thawing the frozen expanded first portion of activated T cells prior to culturing. 
     
     
         4 . The method of  claim 3 , further comprising resting the thawed expanded first portion of the activated T cells prior to culturing. 
     
     
         5 . The method of  claim 1 , wherein the T cells are activated by a stimulator comprising anti-CD3 antibody and an anti-CD28 antibody. 
     
     
         6 . The method of  claim 1 , wherein the period of time is from about 1 day to about 15 days, from about 2 days to about 14 days, from about 3 days to about 13 days, from about 3 days to about 12 days, from about 3 days to about 11 days, from about 3 days to about 10 days, from about 3 days to about 9 days, from about 3 days to about 8 days, from about 3 days to about 7 days, from about 3 days to about 6 days, from about 3 days to about 5 days, from about 3 days to about 4 days, from about 4 days to about 6 days, or from about 4 days to about 5 days after activation. 
     
     
         7 . The method of  claim 1 , wherein the at least one cytokine is selected from the group consisting of interleukin 2 (IL-2), interleukin 7 (IL-7), interleukin 15 (IL-15), and a combination thereof. 
     
     
         8 . The method of  claim 7 , wherein the concentration of IL-2 is from about 10 U/ml to about 500 U/ml, from about 10 U/ml to about 450 U/ml, from about 10 U/ml to about 400 U/ml, from about 10 U/ml to about 350 U/ml, from about 10 U/ml to about 300 U/ml, from about 10 U/ml to about 250 U/ml, from about 10 U/ml to about 200 U/ml, from about 10 U/ml to about 150 U/ml, from about 10 U/ml to about 100 U/ml, from about 10 U/ml to about 50 U/ml, from about 20 U/ml to about 40 U/ml, from about 25 U/ml to about 35 U/ml, or from about 30 U/ml to about 35 U/ml. 
     
     
         9 . The method of  claim 7 , wherein the concentration of IL-7 is from 0.1 ng/ml to 50 ng/ml, from 0.1 ng/ml to 45 ng/ml, from 0.1 ng/ml to 40 ng/ml, from 0.1 ng/ml to 35 ng/ml, from 0.1 ng/ml to 30 ng/ml, from 0.1 ng/ml to 25 ng/ml, from 0.1 ng/ml to 20 ng/ml, from 0.1 ng/ml to 15 ng/ml, from 0.1 ng/ml to 10 ng/ml, from 0.1 ng/ml to 5 ng/ml, from 0.1 ng/ml to 4 ng/ml, from 0.1 ng/ml to 3 ng/ml, from 0.1 ng/ml to 2 ng/ml, from 0.1 ng/ml to 1 ng/ml, or from 0.1 ng/ml to 0.5 ng/ml. 
     
     
         10 . The method of  claim 7 , wherein the concentration of IL-15 is from 0.1 ng/ml to 50 ng/ml, from 0.1 ng/ml to 45 ng/ml, from 0.1 ng/ml to 40 ng/ml, from 0.1 ng/ml to 35 ng/ml, from 0.1 ng/ml to 30 ng/ml, from 0.1 ng/ml to 25 ng/ml, from 0.1 ng/ml to 20 ng/ml, from 0.1 ng/ml to 15 ng/ml, from 0.1 ng/ml to 10 ng/ml, from 0.1 ng/ml to 5 ng/ml, from 0.1 ng/ml to 4 ng/ml, from 0.1 ng/ml to 3 ng/ml, from 0.1 ng/ml to 2 ng/ml, from 0.1 ng/ml to 1 ng/ml, or from 0.1 ng/ml to 0.5 ng/ml. 
     
     
         11 . The method of  claim 1 , wherein the cytokine response is selected from one or more of increased proliferation, reduced apoptosis, increased population of naïve T cells (T N ) and/or stem memory T cells (T scm )/T central memory (T cm ), and a combination thereof. 
     
     
         12 . The method of  claim 4 , wherein the resting is carried out within a period of time from about 0.5 hour to about 48 hours, about 0.5 hour to about 36 hours, about 0.5 hour to about 24 hours, about 0.5 hour to about 18 hours, about 0.5 hour to about 12 hours, about 0.5 hour to about 6 hours, about 1 hour to about 6 hours, about 2 hours to about 5 hours, about 3 hours to about 5 hours, about 4 hours to 6 hours, about 1 hours to about 24 hours, about 2 to about 24 hours, about 12 to about 48 hours, about 0.5 hour to about 120 hours, about 0.5 hour to about 108 hours, about 0.5 hour to about 96 hours, about 0.5 hour to about 84 hours, about 0.5 hour to about 72 hours, or about 0.5 hour to about 60 hours. 
     
     
         13 . The method of  claim 5 , wherein the anti-CD3 antibody and the anti-CD28 antibody each have a concentration of from about 0.1 μg/ml to about 10.0 μg/ml, about 0.1 μg/ml to about 8.0 μg/ml, about 0.1 μg/ml to about 6.0 μg/ml, about 0.1 μg/ml to about 4.0 μg/ml, about 0.1 μg/ml to about 2.0 μg/ml, about 0.1 μg/ml to about 1.0 μg/ml, about 0.1 μg/ml to about 0.8 μg/ml, about 0.1 μg/ml to about 0.6 μg/ml, about 0.1 μg/ml to about 0.5 μg/ml, about 0.1 μg/ml to about 0.25 μg/ml, about 0.2 μg/ml to about 0.5 μg/ml, about 0.2 μg/ml to about 0.3 μg/ml, about 0.3 μg/ml to about 0.5 μg/ml, about 0.3 μg/ml to about 0.4 μg/ml, or about 0.4 μg/ml to about 0.5 μg/ml. 
     
     
         14 . The method of  claim 1 , wherein the obtained T cell is a CD3 +  CD8 +  T cell. 
     
     
         15 . A method of treating a patient having a cancer, comprising administering to the patient an effective amount of the collected expanded second portion of the activated T cells of  claim 1 . 
     
     
         16 . The method of  claim 15 , wherein the T cells are obtained from the patient. 
     
     
         17 . The method of  claim 15 , wherein the T cells are obtained from the healthy donor. 
     
     
         18 . The method of  claim 17 , wherein the cancer is selected from the group consisting of hepatocellular carcinoma (HCC), colorectal carcinoma (CRC), glioblastoma (GB), gastric cancer (GC), esophageal cancer, non-small cell lung cancer (NSCLC), pancreatic cancer (PC), renal cell carcinoma (RCC), benign prostate hyperplasia (BPH), prostate cancer (PCA), ovarian cancer (OC), melanoma, breast cancer, chronic lymphocytic leukemia (CLL), Merkel cell carcinoma (MCC), small cell lung cancer (SCLC), Non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), gallbladder cancer and cholangiocarcinoma (GBC, CCC), urinary bladder cancer (UBC), acute lymphoblastic leukemia (ALL), and uterine cancer (UEC). 
     
     
         19 . A pharmaceutical composition comprising the collected expanded second portion of the activated T cells of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         20 . A T cell population produced by the method of  claim 1 .