Membrane-bound cytokine compositions and methods of modulating an immune response using same

Abstract

The present invention provides a cellular vaccine having a membrane-bound fusion protein that includes a non-antibody immunomodulatory molecule operatively fused to a heterologous membrane attachment domain. Non-antibody immunomodulatory molecules useful in the invention include immunostimulatory and immunosuppressive molecules such as cytokines. In one embodiment, the invention provides a cellular vaccine having a membrane-bound fusion protein that includes a non-antibody immunomodulatory molecule operatively fused to a heterologous membrane attachment domain and, additionally, a disease-associated antigen or immunogenic epitope thereof. Further provided by the invention are methods of modulating an immune response against a disease-associated antigen by administering to an individual a cellular vaccine having a membrane-bound fusion protein that includes a non-antibody immunomodulatory molecule operatively fused to a heterologous membrane attachment domain.

What is claimed is:  
     
         1 . A vaccine, comprising a cell having a membrane-bound fusion protein comprising a non-antibody immunomodulatory molecule operatively fused to a heterologous membrane attachment domain.  
     
     
         2 . The vaccine of  claim 1 , wherein said non-antibody immunomodulatory molecule is an immunostimulatory molecule.  
     
     
         3 . The vaccine of  claim 1 , wherein said non-antibody immunomodulatory molecule is an immunosuppressive molecule.  
     
     
         4 . The vaccine of  claim 1 , wherein said non-antibody immunomodulatory molecule is selected from the group consisting of cytokine and heat shock protein.  
     
     
         5 . The vaccine of  claim 4 , wherein said cytokine is selected from the group consisting of: 
 granulocyte macrophage colony stimulating factor (GM-CSF),    granulocyte colony stimulating factor (G-CSF),    interferon γ (IFN-γ),    interferon α (IFN-α),    tumor necrosis factor-α (TNF-α),    tumor necrosis factor-β (TNF-β),    interleukin-1 (IL-1),    interleukin-2 (IL-2),    interleukin-3 (IL-3),    interleukin-4 (IL-4),    interleukin-6 (IL-6),    interleukin-7 (IL-7),    interleukin-10 (IL-10),    interleukin-12 (IL-12),    lymphotactin and    dendritic cell chemokine 1 (DC-CK1).    
     
     
         6 . The vaccine of  claim 5 , wherein said cytokine is GM-CSF.  
     
     
         7 . The vaccine of  claim 1 , wherein said cell is a prokaryotic cell.  
     
     
         8 . The vaccine of  claim 1 , wherein said cell is a eukaryotic cell.  
     
     
         9 . The vaccine of  claim 8 , wherein said eukaryotic cell is a fibroblast  
     
     
         10 . The vaccine of  claim 8 , wherein said eukaryotic cell is a tumor cell.  
     
     
         11 . The vaccine of  claim 10 , wherein said tumor cell is selected from the group consisting of melanoma cell, renal carcinoma cell, neuroblastoma cell, glioblastoma cell, lung cancer cell, colon tumor cell, breast tumor cell, prostate tumor cell, bladder carcinoma cell and plasmacytoma cell.  
     
     
         12 . The vaccine of  claim 1 , wherein said cell further has a disease-associated antigen or immunogenic epitope thereof.  
     
     
         13 . The vaccine of  claim 12 , wherein said disease-associated antigen is endogenous to said cell.  
     
     
         14 . The vaccine of  claim 12 , wherein said disease-associated antigen is exogenous to said cell.  
     
     
         15 . The vaccine of  claim 12 , wherein said disease-associated antigen is selected from the group consisting of tumor-associated antigen, autoimmune disease-associated antigen, infectious disease-associated antigen, viral antigen, parasitic antigen and bacterial antigen.  
     
     
         16 . The vaccine of  claim 15 , wherein said tumor-associated antigen is selected from the group consisting of p53 and mutants thereof, Ras and mutants thereof, a Bcr/Abl breakpoint peptide, HER-2/neu, HPV E6, HPV E7, carcinoembryonic antigen, MUC-1, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, N-acetylglucosaminyltransferase-V, p15, gp100, MART-1/MelanA, tyrosinase, TRP-1, β-catenin, MUM-1 and CDK-4.  
     
     
         17 . The vaccine of  claim 15 , wherein said autoimmune disease-associated antigen is a T cell receptor derived peptide.  
     
     
         18 . The vaccine of  claim 12 , wherein said disease-associated antigen or immunogenic epitope thereof is operatively fused to said membrane-bound fusion protein.  
     
     
         19 . A method of modulating an immune response against a disease-associated antigen, comprising administering to an individual a vaccine comprising a cell having: 
 (a) a disease-associated antigen or immunogenic epitope thereof and (b) a non-antibody immunomodulatory molecule operatively fused to a heterologous membrane attachment domain.    
     
     
         20 . The method of  claim 19 , wherein said non-antibody immunomodulatory molecule is an immunostimulatory molecule.  
     
     
         21 . The method of  claim 19 , wherein said non-antibody immunomodulatory molecule is an immunosuppressive molecule.  
     
     
         22 . The method of  claim 19 , wherein said non-antibody immunomodulatory molecule is selected from the group consisting of cytokine and heat shock protein.  
     
     
         23 . The method of  claim 22 , wherein said cytokine is selected from the group consisting of GM-CSF, G-CSF, IFN-γ, IFN-α, TNF-α, TNF-β, IL-1. IL-2, IL-3, IL-4, IL-6, IL-7, IL-10, IL-12, lymphotactin and DC-CK1.  
     
     
         24 . The method of  claim 23 , wherein said cytokine is GM-CSF.  
     
     
         25 . The method of  claim 19 , wherein said cell is a prokaryotic cell.  
     
     
         26 . The method of  claim 19 , wherein said cell is a eukaryotic cell.  
     
     
         27 . The method of  claim 26 , wherein said eukaryotic cell is a fibroblast.  
     
     
         28 . The method of  claim 26 , wherein said eukaryotic cell is a tumor cell.  
     
     
         29 . The method of  claim 28 , wherein said tumor cell is selected from the group consisting of melanoma cell, renal carcinoma cell, neuroblastoma cell, glioblastoma cell, lung cancer cell, colon cancer cell, breast cancer cell, prostate cancer cell, bladder carcinoma cell and plasmacytoma cell.  
     
     
         30 . The method of  claim 19 , wherein said disease-associated antigen is endogenous to said cell.  
     
     
         31 . The method of  claim 19 , wherein said disease-associated antigen is exogenous to said cell.  
     
     
         32 . The method of  claim 19 , wherein said disease-associated antigen is selected from the group consisting of a tumor-associated antigen, autoimmune disease-associated antigen, infectious disease-associated antigen, viral antigen, parasitic antigen and bacterial antigen.  
     
     
         33 . The method of  claim 32 , wherein said tumor-associated antigen is selected from the group consisting of p53 and mutants thereof, Ras and mutants thereof, a Bcr/Abl breakpoint peptide, HER-2/neu, HPV E6, HPV E7, carcinoembryonic antigen, MUC-1, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, N-acetylglucosaminyltransferase-V, p15, gp100, MART-1/MelanA, tyrosinase, TRP-1, β-catenin, MUM-1 and CDK-4.  
     
     
         34 . The method of  claim 32 , wherein said autoimmune disease-associated antigen is a T cell receptor derived peptide.  
     
     
         35 . The method of  claim 19 , wherein said disease-associated antigen or immunogenic epitope thereof is operatively fused to said membrane-bound fusion protein.  
     
     
         36 . A nucleic acid molecule, comprising a nucleotide sequence encoding an non-antibody immunomodulatory molecule operatively linked to a heterologous nucleotide sequence encoding a membrane attachment domain functional at neutral or basic pH.  
     
     
         37 . The nucleic acid molecule of  claim 36 , wherein said non-antibody immunomodulatory molecule is an immunostimulatory molecule.  
     
     
         38 . The nucleic acid molecule of  claim 36 , wherein said non-antibody immunomodulatory molecule is an immunosuppressive molecule.  
     
     
         39 . The nucleic acid molecule of  claim 36 , wherein said non-antibody immunomodulatory molecule is selected from the group consisting of cytokine and heat shock protein.  
     
     
         40 . The nucleic acid molecule of  claim 39 , wherein said cytokine is selected from the group consisting of GM-CSF, G-CSF, IFN-γ, IFN-α, TNF-α, TNF-β, IL-1. IL-2, IL-3, IL-4, IL-6, IL-7, IL-10, IL-12, lymphotactin and DC-CK1.  
     
     
         41 . The nucleic acid molecule of  claim 40 , wherein said cytokine is GM-CSF.  
     
     
         42 . The nucleic acid molecule of  claim 36 , further comprising an operatively linked nucleotide sequence encoding a disease-associated antigen or immunogenic epitope thereof.  
     
     
         43 . The nucleic acid molecule of  claim 42 , wherein said disease-associated antigen is selected from the group consisting of tumor-associated antigen, autoimmune disease-associated antigen, infectious disease associated antigen, viral antigen, parasitic antigen and bacterial antigen.  
     
     
         44 . The nucleic acid molecule of  claim 43 , wherein said tumor-associated antigen is selected from the group consisting of p53 and mutants thereof, Ras and mutants thereof, Bcr/Abl breakpoint peptides, HER-2/neu, HPV E6, HPV E7, carcinoembryonic antigen, MUC-1, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, N-acetylglucosaminyltransferase-V, p15, gp100, MART-1/MelanA, tyrosinase, TRP-1, β-catenin, MUM-1 and CDK-4.  
     
     
         45 . The nucleic acid molecule of  claim 43 , wherein said autoimmune disease-associated antigen is a T cell receptor derived peptide.  
     
     
         46 . A nucleic acid molecule, comprising a nucleotide sequence encoding a non-antibody immunomodulatory molecule operatively linked to a heterologous nucleotide sequence encoding a membrane attachment domain, provided that said membrane attachment domain is not derived from diphtheria toxin.