Recombinant influenza viruses expressing tumor-associated antigens as antitumor agents

Abstract

The present invention relates to the engineering of recombinant influenza viruses that express tumor-associated antigens. Expression of tumor-associated antigens by these viruses can be achieved by engineering specific epitopes into influenza virus proteins, or by engineering viral genes that encode a viral protein and the specific antigen as independent polypeptides. Tumor-bearing patients can be immunized with the recombinant influenza viruses alone, or in combination with another treatment, to induce an immune response that leads to tumor reduction. The recombinant viruses can also be used to vaccinate high risk tumor-free patients to prevent tumor formation in vivo.

1 . A recombinant influenza virus the genome of which contains a region encoding a tumor antigen or an epitope of a tumor antigen.  
     
     
         2 - 19 . (canceled).  
     
     
         20 . The recombinant influenza virus of  claim 1  in which the region is inserted into an open reading frame of a genomic segment of the influenza virus.  
     
     
         21 . The recombinant influenza virus of  claim 1  in which the region is inserted into an open reading frame of a genomic segment of the influenza virus.  
     
     
         22 . The recombinant influenza virus of  claim 20  or  21 , wherein said genomic segment is a structural gene of the influenza virus.  
     
     
         23 . The recombinant influenza virus of  claim 22 , wherein said structural gene is hemagglutinin (HA) or neuraminadase (NA).  
     
     
         24 . The recombinant influenza virus of  claim 1 , wherein said tumor antigen is a melanocyte tumor antigen, a widely shared antigen, a mutated antigen or a nonmelanoma antigen.  
     
     
         25 . The recombinant influenza virus of  claim 24 , wherein said melanocyte tumor antigen is gp100, MART-1/MelanA, TRP-1 (gp75) or tyrosinase.  
     
     
         26 . The recombinant influenza virus of  claim 24 , wherein said widely shared antigen is MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, N-acetylglucosaminyltransferase-V or p15.  
     
     
         27 . The recombinant influenza virus of  claim 24 , wherein said mutated antigen is β-catenin, MUM-1, or CDK4.  
     
     
         28 . The recombinant influenza virus of  claim 24 , wherein said nonmelanoma antigen is a breast, ovarian, cervical or pancreatic antigen.  
     
     
         29 . The recombinant influenza virus of  claim 24 , wherein the nonmelanoma antigen is Her-2/neu, human papillomavirus-E7 or MUC-1.  
     
     
         30 . An immunogenic composition comprising an effective amount of the recombinant influenza virus of  claim 1  or  24 , and a pharmaceutically acceptable carrier.  
     
     
         31 . The immunogenic composition of  claim 30  in which the recombinant influenza virus is a live virus.  
     
     
         32 . The immunogenic composition of  claim 31  in which the recombinant influenza virus is inactivated.  
     
     
         33 . The immunogenic composition of  claim 32  further comprising an adjuvant.  
     
     
         34 . A method of inducing an immune response against tumor cells, said method comprising administering to a tumor-bearing patient or a tumor-free patient an effective amount of the immunogenic formulation of  claim 30 .  
     
     
         35 . The method of  claim 34  further comprising the subsequent administration of a booster preparation.  
     
     
         36 . The method of  claim 35 , wherein the booster preparation comprises a subunit vaccine containing the same tumor antigen or the same epitope of the tumor antigen, or a different serotype than the recombinant influenza virus used in the initial immunization, or a recombinant virus other than an influenza virus engineered to express the same tumor antigen or the same epitope of the tumor antigen.  
     
     
         37 . The method of  claim 34  further comprising administering to said tumor-bearing host chemotherapy, radiation therapy or a bone marrow transplant, or conducting surgery on said tumor-bearing patient.  
     
     
         38 . The method of  claim 34 , wherein the immunogenic composition is administered to the tumor-bearing host orally, intradermally, intramuscularly, intraperitoneally, intravenously, subcutaneously, intranasally, transdermally, epidurally, pulmonary, gastrically, intestinally, rectally, vaginally, or urethrally.  
     
     
         39 . The method of  claim 34 , wherein said patient is a mammal.  
     
     
         40 . The method of  claim 34 , wherein said patient is a human.  
     
     
         41 . A method for activating T lymphocytes, said method comprising contacting T lymphocytes with the recombinant influenza virus of  claim 1 .  
     
     
         42 . A method for generating dendritic cells expressing a tumor antigen or an epitope of a tumor antigen, said method comprising contacting dendritic cells with the recombinant influenza virus of  claim 1 .  
     
     
         43 . A method for activating T lymphocytes, said method comprising a) contacting isolated T lymphocytes with the recombinant influenza virus of  claim 1;  and b) expanding the T lymphocytes.  
     
     
         44 . A method for activating T lymphocytes comprising contacting the dendritic cells produced by the method of  claim 42  with T lymphocytes.  
     
     
         45 . A method of inducing an immune response against tumor cells, said method comprising administering to a tumor-bearing patient or a tumor-free patient an effective amount of the activated T lymphocytes produced by the method of  claim 43  or  44 , wherein the activated T lymphocytes are histocompatible with the patient.  
     
     
         46 . A method of inducing an immune response against tumor cells, said method comprising administering to a tumor-bearing patient or a tumor-free patient an effective amount of the dendritic cells produced by the methods of  claim 42 , wherein the dendritic cells are histocompatible with the patient.