Multiple antigen specific cell therapy methods

Abstract

The present invention provides methods of preparing a population of activated T cells by co-culturing T cells with dendritic cells loaded with a plurality of tumor antigen peptides. Also provided are methods of treating cancer in an individual using the activated T cells, pharmaceutical compositions and kits for cell-based cancer immunotherapy.

1 . A method of preparing a population of activated T cells, the method comprising:
 a) contacting a population of dendritic cells with a plurality of tumor antigen peptides to obtain a population of dendritic cells loaded with the plurality of tumor antigen peptides;   b) co-culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides and a population of T cells in an initial co-culture medium comprising a plurality of cytokines and an immune checkpoint inhibitor to provide a co-culture; and   c) adding an anti-CD3 antibody to the co-culture at about 3 to 7 days after the co-culturing starts, thereby obtaining the population of activated T cells.   
     
     
         2 . The method of  claim 1 , wherein step a) further comprises culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides in a DC maturation medium comprising a toll-like receptor (TLR) agonist. 
     
     
         3 . The method of  claim 2 , wherein the TLR agonist is selected from the group consisting of monophosphoryl lipid (MPLA), Poly I:C, resquimod, gardiquimod, and CL075. 
     
     
         4 . A method of preparing a population of activated T cells, the method comprising:
 a) contacting a population of dendritic cells with a plurality of tumor antigen peptides to obtain a population of dendritic cells loaded with the plurality of tumor antigen peptides;   b) culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides in a DC maturation medium comprising MPLA; and   c) co-culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides and a population of T cells, thereby obtaining the population of activated T cells.   
     
     
         5 . The method of  claim 2 , wherein the DC maturation medium comprises interferon-γ (INFγ), MPLA, and prostaglandin E2 (PGE2). 
     
     
         6 - 7 . (canceled) 
     
     
         8 . The method of  claim 5 , wherein the MPLA is present in the DC maturation medium at a concentration of at least about 0.5 μg/mL. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 5 , wherein step c) comprises: co-culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides and a population of T cells in an initial co-culture medium comprising a plurality of cytokines and an immune checkpoint inhibitor to provide a co-culture; and adding an anti-CD3 antibody to the co-culture, t hereby obtaining the population of activated T cells. 
     
     
         11 . The method of  claim 1 , wherein the plurality of cytokines comprises IL-2, IL-7, IL-15 and IL-21. 
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 1 , wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. 
     
     
         14 . (canceled) 
     
     
         15 . The method of  claim 1 , wherein the anti-CD3 antibody is added to the co-culture at about 3 to 7 days after the co-culturing starts. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 1 , wherein the population of dendritic cells loaded with the plurality of tumor antigen peptides and the population of T cells are co-cultured for at least about 10 days in the presence of the anti-CD3 antibody. 
     
     
         18 . (canceled) 
     
     
         19 . The method of  claim 1 , wherein the population of dendritic cells is obtained by inducing differentiation of a population of monocytes from PBMCs. 
     
     
         20 . The method of  claim 1 , wherein the population of dendritic cells and the population of T cells are obtained from the same individual. 
     
     
         21 . The method of  claim 1 , wherein the plurality of tumor antigen peptides comprises a neoantigen peptide. 
     
     
         22 . An isolated population of activated T cells prepared using the method of  claim 1 . 
     
     
         23 . A method of treating a cancer in an individual, comprising:
 a) contacting a population of dendritic cells with a plurality of tumor antigen peptides to obtain a population of dendritic cells loaded with the plurality of tumor antigen peptides;   b) co-culturing the population of dendritic cells loaded with the plurality of tumor antigen peptides and a population of T cells in an initial co-culture medium comprising a plurality of cytokines and an immune checkpoint inhibitor to provide a co-culture;   c) adding an anti-CD3 antibody to the co-culture at about 3 to 7 days after the co-culturing starts, thereby obtaining activated T cells; and   d) administering to the individual an effective amount of the activated T cells of  claim 22 .   
     
     
         24 - 32 . (canceled)