Antitumor vaccination using allogeneic tumor cells expressing alpha (1,3)-galactosyltransferase

Abstract

The invention relates to methods and compositions for causing the selective targeting and killing of tumor cells. Through ex vivo gene therapy protocols tumor cells are engineered to express an α(1,3)galactosyl epitope. The cells are then irradiated or otherwise killed and administered to a patient. The α-galactosyl epitope causes opsonization of the tumor cell enhancing uptake of the opsonized tumor cell by antigen presenting cells which results in enhanced tumor specific antigen presentation. The animal's immune system thus is stimulated to produce tumor specific cytotoxic cells and antibodies which will attack and kill tumor cells present in the animal.

1 . A pharmaceutical composition for killing of lung tumor cells in an animal by stimulating an immune response to said cells comprising: 
 a mixture of attenuated vaccine tumor cells, said cells comprising an α(1,3)galactosyltransferase encoding polynucleotide sequence and a plurality of cell surface glycoproteins or glycolipids on which an α-Gal epitope is present, and a carrier.    
     
     
         2 . The composition of  claim 1  wherein said attenuated vaccine tumor cells lung cancer cell lines derived from the group consisting of: small cell lung carcinoma, large cell carcinoma, squamous cell carcinoma, lung adenocarcinoma, bronchoalveolar carcinoma.  
     
     
         3 . The composition of  claim 2  wherein said vaccine tumor cells are selected from the group consisting of: cell lines A549, NCI-H460 and NCI-H520 cells.  
     
     
         4 . The composition of  claim 1  wherein said pharmaceutical composition comprises the cell lines HAL1, HAL2 and/or HAL3.  
     
     
         5 . The composition of  claim 1  further comprising an adjuvant.  
     
     
         6 . The composition of  claim 1  where said vaccine tumor cells are attenuated by gamma irradiation.  
     
     
         7 . A pharmaceutical composition for stimulating an immune response to tumor antigens present in lung tumor cells, regardless of their glycosylation state, comprising: 
 a mixture of attenuated vaccine tumor cells, said cells expressing an α(1,3)galactosyltransferase protein and a plurality of cell surface glycoproteins or glycolipids on which an α-Gal epitope is present, and a suitable carrier.    
     
     
         8 . The composition of  claim 7  wherein said attenuated vaccine tumor cells lung cancer cell lines derived from the group consisting of: small cell lung carcinoma, large cell carcinoma, squamous cell carcinoma, lung adenocarcinoma, bronchoalveolar carcinoma.  
     
     
         9 . The composition of  claim 8  wherein said vaccine tumor cells are selected from the group consisting of: cell lines A549, NCI-H460 and NCI-H520 cells.  
     
     
         10 . The composition of  claim 7  wherein said pharmaceutical composition comprises the cell lines HAL1, HAL2 and/or HAL3.  
     
     
         11 . The composition of  claim 7  further comprising an adjuvant.  
     
     
         12 . The composition of  claim 7  where said vaccine tumor cells are attenuated by gamma irradiation.  
     
     
         13 . A pharmaceutical composition for inducing an immune response against lung tumor cells in human subjects comprising: 
 a mixture of attenuated vaccine tumor cells, said cells comprising an α(1,3)galactosyltransferase encoding sequence and a plurality of cell surface glycoproteins or glycolipids on which an α-Gal epitope is present, said vaccine tumor cells selected from the group consisting of A549, NCI-H460, NCI-H520; and a carrier.    
     
     
         14 . A pharmaceutical composition for treating lung tissue tumor cells in human subjects comprising one or more cell lines selected from the group of HAL1, HAL2 or HAL3.  
     
     
         15 . A method of making a therapeutic pharmaceutical composition for inducing an immune response against lung tumor cells in humans, comprising: 
 obtaining a collection of live lung carcinoma cells,    introducing to said cells a nucleotide sequence which encodes upon expression α(1,3)galactosyltransferase, so that said α(1,3)galactosyltransferase synthesizes an α-galactosyl epitope on a plurality of cell surface glycoproteins on said cells,    collecting said lung tumor cells to form a mixture of cells with a plurality of glycoproteins having an α-galactosyl epitope and    attenuating said cells by exposing them to gamma irradiation.    
     
     
         16 . The method of  claim 15  wherein said cells are allogeneic.  
     
     
         17 . The method of  claim 15  wherein said cells are syngeneic.  
     
     
         18 . The method of  claim 15  wherein said cells are autologous.  
     
     
         19 . The method of  claim 15  wherein said α(1,3)galactosyltransferase encoding sequence is a murine sequence.  
     
     
         20 . The method of  claim 15  wherein said step of introducing said α(1,3)galactosyltransferase encoding polynucleotide sequence is mediated by a vector.  
     
     
         21 . The method of  claim 20  where the vector is a plasmid vector  
     
     
         22 . The method of  claim 20  where the vector is a viral vector  
     
     
         23 . The method of  claim 22  where the viral vector is selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, an adeno-associated vector and a herpes simplex virus vectors.  
     
     
         24 . A therapeutic pharmaceutical composition produced by the method of  claim 15 .  
     
     
         25 . A method of inducing immune mediated destruction of lung tumor cells in human patients comprising: 
 administering to a human patient a mixture of attenuated vaccine lung tumor cells, said cells comprising a plurality of cell surface glycoproteins on which an α-Gal epitope is present.    
     
     
         26 . A method of treating human subjects with lung tumors comprising: 
 administering to said subject a therapeutically effective amount of a composition comprising a mixture of attenuated vaccine tumor cells genetically modified with a polynucleotide that encodes an α(1,3)galactosyltransferase protein, said cells selected from the group consisting of autologous, syngeneic, and allogeneic lung tumor cells.    
     
     
         27 . The method of  claim 26  wherein said α(1,3)galactosyltransferase polynucleotide is of murine origin.  
     
     
         28 . The method of  claim 26  wherein said polynucleotide encodes an α(1,3)galactosyltransferase with at least about 70% amino acid sequence identity or 85% amino acid sequence similarity to that of SEQ ID NO: 2.  
     
     
         29 . The method of  claim 26  where the α(1,3)galactosyltransferase gene is introduced into the cells by a vector.  
     
     
         30 . The method of  claim 29  where the vector is a viral vector.  
     
     
         31 . The method of  claim 30  where the viral vector is based on murine retroviral vector, lentiviral vectors, adenoviral vectors, adeno-associated vector and herpes simplex virus vectors.  
     
     
         32 . A method of treating pre-established lung tumors in a human patient, comprising, introducing to said human patient a therapeutically effective dose of lethally irradiated allogeneic, syngeneic or autologous lung tumor cells that express the α(1,3)galactosyltransferase gene.  
     
     
         33 . The method of  claim 32  where the α(1,3)galactosyltransferase polynucleotide is of murine origin.  
     
     
         34 . The method of  claim 33  wherein said gene encodes an α(1,3)galactosyltransferase with at least about 70% amino acid sequence identity or 85% amino acid sequence similarity to that of SEQ ID NO: 2.  
     
     
         35 . The method of  claim 34  where the α(1,3)galactosyltransferase gene is introduced into the cells by a vector.  
     
     
         36 . The method of  claim 35  where the vector is a viral vector.  
     
     
         37 . The method of  claim 36  where the viral vector is based on murine retroviral vector, lentiviral vectors, adenoviral vectors, adeno-associated vector and herpes simplex virus vectors.  
     
     
         38 . A pharmaceutical composition for stimulating an immune response against lung tumor cells in human subjects comprising a mixture of the lethally gamma-irradiated vaccine cell lines HAL1, HAL2 and HAL3, said cells being derived from the lung cancer cell lines A549, NCI-H460 and NCI-H520 by transduction with an pLNCKG retroviral vector, which encodes the murine α(1,3)galactosyltransferase gene under the control of the cytomegalovirus promoter.  
     
     
         39 . A method to induce an immune response against lung tumor cells in human subjects comprising: administering a preventive or therapeutic effective dose of a mixture of the vaccine cell lines HAL1, HAL2 and HAL3, said cells being derived from the lung cancer cell lines A549, NCI-H460 and NCI-H520 by transduction with the retroviral vector pLNCKG, wherein said vector encodes the murine α(1,3)galactosyltransferase gene under the control of the cytomegalovirus promoter; and further wherein said vaccine cells have been attenuated by gamma irradiation prior to administration to the patients.