Methods for manufacturing t cells

Abstract

The disclosure relates to methods of manufacturing T cells for adoptive immunotherapy. The disclosure further provides for methods of genetically transducing T cells, methods of using T cells, and T cell populations thereof. In an aspect, the disclosure provides for methods of thawing frozen peripheral blood mononuclear cells (PBMC), resting the thawed PBMC, activating the T cell in the cultured PBMC with an anti-CD3 antibody and an anti-CD28 antibody immobilized on a solid phase, transducing the activated T cell with a viral vector, expanding the transduced T cell, and obtaining expanded T cells.

What is claimed is: 
     
         1 . A method of transducing a T cell population comprising
 thawing frozen peripheral blood mononuclear cells (PBMC),   resting the thawed PBMC,   activating the T cells in the rested PBMC with an anti-CD3 antibody and anti-CD28 antibody,   transducing the activated T cells with a viral vector,   expanding the transduced T cells, and   obtaining the expanded T cells,
 wherein the expanded T cells are capable of specifically binding a peptide consisting of the amino acid sequence of SLLMWITQC (SEQ ID NO: 131) or GVYDGREHTV (SEQ ID NO: 89). 
   
     
     
         2 . The method of  claim 1 , wherein the activation further comprises incubation with IL-2. 
     
     
         3 . The method of  claim 1 , wherein the IL-2 concentration is between about 50 IU/mL and 150 IU/mL. 
     
     
         4 . The method of  claim 1 , wherein the viral vector is a lentivirus vector. 
     
     
         5 . The method of  claim 1 , wherein the T cells are expanded for 1-15 days. 
     
     
         6 . The method of  claim 1 , wherein the T cells are expanded in the presence of IL-2, IL-7, IL-12, IL-15, or a combination thereof. 
     
     
         7 . The method of  claim 1 , wherein the T cells are CD4+. 
     
     
         8 . The method of  claim 1 , wherein the T cells are CD8+. 
     
     
         9 . A method of treating a patient having a cancer comprising administering a composition comprising the T cell of  claim 1 . 
     
     
         10 . The method of  claim 9 , wherein the cancer is melanoma, ovarian cancer, esophageal cancer, non-small cell lung cancer (NSCLC), or a combination thereof. 
     
     
         11 . The method of  claim 9 , the T cells are autologous. 
     
     
         12 . The method of  claim 9 , wherein the patient is HLA-A*02. 
     
     
         13 . The method of  claim 9 , wherein the dosage of the T-cells is about 1×10 6  to about 1×10 9  transduced T cells/m 2  (or kg) of the patient. 
     
     
         14 . The method of  claim 9 , wherein the T-cells are administered via continuous infusion. 
     
     
         15 . The method of  claim 1 , wherein the expanded T cells are capable of specifically binding a peptide consisting of the amino acid sequence of SLLMWITQC (SEQ ID NO: 131). 
     
     
         16 . The method of  claim 15 , wherein the viral vector comprises a nucleic acid encoding a T cell receptor (TCR) that binds a peptide consisting of the amino acid sequence of SLLMWITQC (SEQ ID NO: 131). 
     
     
         17 . The method of  claim 1 , wherein the expanded T cells are capable of specifically binding a peptide consisting of the amino acid sequence of GVYDGREHTV (SEQ ID NO: 89). 
     
     
         18 . The method of  claim 17 , wherein the viral vector comprises a nucleic acid encoding a TCR that binds a peptide consisting of the amino acid sequence of GVYDGREHTV (SEQ ID NO: 89). 
     
     
         19 . The method of  claim 6 , wherein the T cells are expanded in the presence of IL-7. 
     
     
         20 . The method of  claim 6 , wherein the T cells are expanded in the presence of IL-15.