Combination immunogene therapy

Abstract

The present invention provides to immunogene therapy protocols for the treatment of tumors. In particular, the present invention provides combinations of immune-modulating proteins that induce systemic immunity against tumors. In addition, the present invention provides humanized animal models suitable for the evaluation of anti-human tumor immunity and permit the identification of combinations of immune-modulating genes which when delivered to human tumor cells induce an effective anti-tumor response, including a systemic anti-tumor response.

I claim:  
     
         1 . An immunodeficient mouse comprising: 
 a) human T lymphocytes expressing the CD45 antigen, wherein at least 5% of the human T cells expressing the CD45 antigen represent immature naive T lymphocytes; and    b) human tumor cells;    wherein said immunodeficient mouse is a SCID/beige mouse.    
     
     
         2 . The mouse according to  claim 1 , wherein said tumor cells are from a tumor cell line.  
     
     
         3 . The mouse according to  claim 1 , wherein said tumor cells are from a primary tumor.  
     
     
         4 . The mouse according to  claim 1 , wherein said tumor cells are derived from central nervous system cells.  
     
     
         5 . The mouse according to  claim 4 , wherein said tumor cells derived from central nervous system cells are glioblastoma cells.  
     
     
         6 . The mouse according to  claim 1 , wherein at least one of said tumor cells contains at least one transgene.  
     
     
         7 . The mouse according to  claim 6 , wherein at least one of said transgenes is a human immunomodulator gene.  
     
     
         8 . The mouse according to  claim 6 , wherein at least one of said transgenes is delivered by a viral vector.  
     
     
         9 . The mouse according to  claim 1 , further comprising an immunogen.  
     
     
         10 . The mouse according to  claim 9 , wherein said immunogen is a vaccine.  
     
     
         11 . A tumor cell vaccine comprising a tumor cell expressing B7-2 and at least one additional immune modulator.  
     
     
         12 . The vaccine according to  claim 11 , wherein said at least one additional immune modulator is a cytokine.  
     
     
         13 . The vaccine according to  claim 12 , wherein said cytokine is selected from the group consisting of interleukin 2, interleukin 4, interleukin 6, interleukin 7, interleukin 12, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, interferon-gamma, tumor necrosis factor-alpha.  
     
     
         14 . A method of treating a tumor comprising: 
 a) providing: 
 i) a subject having a tumor of the central nervous system;  
 ii) an expression vector encoding the human B7-2 protein and at least one additional immune modulator;  
   b) transferring said expression vector into said tumor under conditions such that said B7-2 protein and said immune-modulator are expressed by at least a portion of said tumor.    
     
     
         15 . The method according to  claim 14  further comprising, prior to transfer of said expression vector, the step of removing at least a portion of said tumor from said subject and following said transfer of said expression vector, irradiating said tumor cells expressing said B7-2 protein and said immune-modulator and introducing said irradiated tumor cells back into said subject to create an immunized subject.  
     
     
         16 . The method according to  claim 15  further comprising, introducing at least one additional dose of irradiated tumor cells expressing said B7-2 protein and said immune-modulator into said immunized subject.  
     
     
         18 . The tumor cell composition according to claim  17 , wherein said at least one additional immune modulator is a cytokine protein.  
     
     
         19 . The tumor cell composition according to  claim 18 , wherein said cytokine protein is selected from the group consisting of interleukin 2, interleukin 4, interleukin 6, interleukin 7, interleukin 12, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, interferon-gamma, and tumor necrosis factor-alpha.  
     
     
         20 . The tumor cell composition according to  claim 18 , wherein said cytokine protein is granulocyte-macrophage colony stimulating factor.  
     
     
         21 . An expression vector comprising a polynucleotide sequence encoding a B7-2 protein and at least one additional immune modulating protein, or a functional fragment of said B7-2 protein or said immune modulator.  
     
     
         22 . The expression vector according to  claim 21 , wherein said at least one additional immune modulating protein is a cytokine protein.  
     
     
         23 . The expression vector according to  claim 22 , wherein said cytokine protein is selected from the group consisting of interleukin 2, interleukin 4, interleukin 6, interleukin 7, interleukin 12, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, interferon-gamma, and tumor necrosis factor-alpha.  
     
     
         24 . The expression vector according to  claim 22 , wherein said cytokine protein is granulocyte-macrophage colony stimulating factor.  
     
     
         25 . The expression vector according to  claim 21 , wherein said expression vector is a viral vector.  
     
     
         26 . The expression vector according to  claim 25 , wherein said viral vector is a retroviral vector.  
     
     
         27 . The expression vector according to  claim 25 , wherein said viral vector is an adenoviral vector.  
     
     
         28 . The expression vector according to  claim 21 , wherein said expression vector is encapsulated by, or complexed with, a liposome.  
     
     
         29 . A method for the treatment or prevention of cancer comprising: 
 a) providing a polynucleotide encoding a B7-2 protein and at least one additional immune modulator, or a functional fragment of said B7-2 protein or said immune modulator;    b) transferring said polynucleotide into cancer cells under conditions such that said B7-2 protein and said immune modulator are expressed by at least a portion of said cancer cells; and    c) administering an effective amount of the modified cancer cells of step b) to a patient.    
     
     
         30 . The method according to  claim 29  further comprising irradiating said cancer cells expressing said B7-2 protein and said immune modulator prior to administering said irradiated cancer cells into said patient.  
     
     
         31 . The method according to  claim 30 , further comprising introducing at least one additional dose of irradiated cancer cells expressing said B7-2 protein and said immune modulator into said immunized subject.  
     
     
         32 . The method according to  claim 29 , wherein said at least one additional immune modulator is a cytokine protein.  
     
     
         33 . The method according to  claim 32 , wherein said cytokine protein is selected from the group consisting of interleukin 2, interleukin 4, interleukin 6, interleukin 7, interleukin 12, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, interferon-gamma, and tumor necrosis factor-alpha.  
     
     
         34 . The method according to  claim 32 , wherein said cytokine protein is granulocyte-macrophage colony stimulating factor.  
     
     
         35 . The method according to  claim 29 , wherein said polynucleotide is transferred by a viral vector.  
     
     
         36 . The method according to  claim 35 , wherein said viral vector is a retroviral vector.  
     
     
         37 . The method according to  claim 35 , wherein said viral vector is an adenoviral vector.  
     
     
         38 . The method according to  claim 29 , wherein said polynucleotide is encapsulated by, or complexed with, a liposome.  
     
     
         39 . The method according to  claim 29 , wherein said cancer cells are from a solid tumor.  
     
     
         40 . The method according to  claim 29 , wherein said cancer cells are from a brain tumor.  
     
     
         41 . The method according to  claim 40 , wherein said brain tumor is a glioblastoma.  
     
     
         42 . The method according to  claim 29 , wherein said cancer cells are from a melanoma.  
     
     
         43 . A method for the treatment or prevention of cancer comprising administering to a subject in need thereof an effective amount of a tumor vaccine comprising a tumor cell modified to express a B7-2 protein and at least one additional immune modulator, or a functional fragment of said B7-2 protein or said immune modulator.  
     
     
         44 . The method according to  claim 43 , wherein said at least one additional immune modulator is a cytokine protein.  
     
     
         45 . The method according to  claim 44 , wherein said cytokine protein is selected from the group consisting of interleukin 2, interleukin 4, interleukin 6, interleukin 7, interleukin 12, granulocyte-macrophage colony stimulating factor, granulocyte colony stimulating factor, interferon-gamma, and tumor necrosis factor-alpha.  
     
     
         46 . The method according to  claim 43 , wherein said cytokine protein is granulocyte-macrophage colony stimulating factor.  
     
     
         47 . The method according to  claim 43 , wherein said cancer cells are from a tumor.  
     
     
         48 . The method according to  claim 43 , wherein said cancer cells are from a brain tumor.  
     
     
         49 . The method according to  claim 48 , wherein said brain tumor is a glioblastoma.  
     
     
         50 . The method according to  claim 43 , wherein said cancer cells are from a melanoma.