Universal GM-CSF Expressing Bystander Human Cell Line

Abstract

The present invention provides a universal immunomodulatory cytokine-expressing bystander cell line, a composition comprising such a cell line and a cancer antigen, a method of making such a cell line, and a method of using such a composition.

1 . A universal bystander cell line, which: (i) is a human cell line modified to lack major histocompatibility class I (MHC-I) antigens and major histocompatibility class II (MHC-II) antigens, and (ii) is modified by introduction of a nucleic acid molecule comprising a nucleic acid sequence encoding granulocyte macrophage-colony stimulating factor (GM-CSF) operably linked to a promoter, wherein the universal bystander cell line expresses about 500 ng or greater GM-CSF/10 6  cells/24 hours. 
     
     
         2 . The universal bystander cell line of  claim 1 , which expresses about 1,000 ng or greater GM-CSF/10 6  cells/24 hours. 
     
     
         3 . The universal bystander cell line of  claim 1 , wherein the promoter is the human cytomegalovirus promoter. 
     
     
         4 . A composition comprising the universal bystander cell line of  claim 3  and a cancer antigen. 
     
     
         5 . A method of stimulating an immune response to a cancer in a human patient, which method comprises administering to said patient the composition of  claim 4 , wherein said cancer antigen is an antigen of said cancer and wherein said composition is irradiated, whereupon administration of said composition, an immune response to said cancer is stimulated. 
     
     
         6 . The method of  claim 5 , wherein said cancer antigen is a cell of said cancer. 
     
     
         7 . In a method of cancer immunotherapy, the improvement comprising administering to a human patient having a cancer the composition of  claim 4 , wherein said cancer antigen is an antigen of said cancer and wherein said composition is irradiated. 
     
     
         8 . The universal bystander cell line of  claim 1 , which grows in defined medium. 
     
     
         9 . The universal bystander cell line of  claim 1 , wherein said promoter is a cytomegalovirus promoter. 
     
     
         10 . The universal bystander cell line of  claim 3 , wherein said nucleic acid molecule further comprises a nucleic acid sequence encoding hygromycin resistance operably linked to a promoter and said universal bystander cell line is selected by growth in a culture medium comprising about 400 μg/ml or greater hygromycin. 
     
     
         11 . The universal bystander cell line of  claim 10 , wherein said universal bystander cell line is selected by growth in a culture medium comprising about 1,000 μg/ml or greater hygromycin. 
     
     
         12 . A composition comprising the universal bystander cell line of  claim 1  and a cancer antigen. 
     
     
         13 . A method of stimulating an immune response to a cancer in a human patient, which method comprises administering to said patient the composition of  claim 12 , wherein said cancer antigen is an antigen of said cancer and wherein said composition is irradiated, whereupon administration of said composition, an immune response to said cancer is stimulated. 
     
     
         14 . The method of  claim 13 , wherein said cancer antigen is a cell of said cancer. 
     
     
         15 . In a method of cancer immunotherapy, the improvement comprising administering to a human patient having a cancer the composition of  claim 12 , wherein said cancer antigen is an antigen of said cancer and wherein said composition is irradiated.