Nucleotide sequence expressing an exosome-anchoring protein for use as vaccine

Abstract

The present invention concerns a nucleotide sequence expressing a fusion protein, said fusion protein comprising or consisting of an exosome-anchoring protein fused at its C-terminus with an antigen, or a DNA expression vector comprising said nucleotide sequence, for use as vaccine.

1 . Nucleotide sequence expressing a fusion protein, said fusion protein comprising or consisting of the exosome-anchoring protein of sequence SEQ ID NO: 1 fused at its C-terminus with an immunogenic antigen, or a DNA expression vector comprising said nucleotide sequence, for use in vaccine prevention and therapy by inducing a CTL immune response, wherein SEQ ID NO: 1 is the following sequence: 
       
         
           
                 
                 
               
                     
                   MGCKWSKSSV VGWPAVRERM RRAEPAADGV GAASRDLEKH 
                 
                     
                     
                 
                     
                   GAITSSNTAA TNADCAWLEA QEEEEVGFPV TPQVPLRPMT  
                 
                     
                     
                 
                     
                   YKAAVDLSHF LKEKGGLEGL IHSQRRQDIL DLWIYHTQGY 
                 
                     
                     
                 
                     
                   FPDWQNYPTG PGIRYPLTFG WCYKLVPVEP EKLEEANKGE 
                 
                     
                     
                 
                     
                   NTSLLHPVSL HGMDDPGREV LEWRFDSRLA FHHVARELHP 
                 
                     
                     
                 
                     
                   EYFKNC. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . Nucleotide sequence or DNA expression vector according to  claim 1 , for use according to  claim 1 , wherein said nucleotide sequence or DNA expression vector is administered by intramuscular administration. 
     
     
         3 . Nucleotide sequence or DNA expression vector according to  claim 1 , for use according to  claim 1 , wherein the antigen is chosen from the group consisting of Human Papilloma virus antigen such as E6 and E7, HIV antigen such as Gag and Tat, Ebola virus antigen such as VP24, VP40, NP, and GP, West Nile virus antigen such as NS3, HBV antigen such as Core, HCB antigen such as Core, NS3, E1 and E2, Crimean-Congo virus antigen such as GP and NP, Influenza A virus antigen such as NP and M1, human melanoma antigen such as MAGE-A3 and MART-1, human tumor-associated antigens such as Her2/Neu, Hox B7. 
     
     
         4 . Nucleotide sequence or DNA expression vector according to  claim 1 , for use according to  claim 1 , wherein the nucleotide sequence expressing the exosome-anchoring protein of sequence SEQ ID NO:1 is the following nucleotide sequence SEQ ID NO:2: 
       
         
           
                 
               
                   Atg ggt tgc aag tgg tca aaa agt agt gtg gtt gga  
                 
                     
                 
                   tgg cct gct gta agg gaa aga atg aga cga gct gag  
                 
                     
                 
                   cca gca gca gat ggg gtg gga gca gca tct cga gac  
                 
                     
                 
                   cta gaa aaa cat gga gca atc aca agt agc aat aca  
                 
                     
                 
                   gca gct acc aat gct gat tgt gcc tgg cta gaa gca 
                 
                     
                 
                   caa gag gag gag gag gtg ggt ttt cca gtc aca cct  
                 
                     
                 
                   cag gta cct tta aga cca atg act tac aag gca gct 
                 
                     
                 
                   gta gat ctt agc cac ttt tta aaa gaa aag ggg gga  
                 
                     
                 
                   ctg gaa ggg cta att cac tcc caa cga aga caa gat 
                 
                     
                 
                   atc ctt gat ctg tgg atc tac cac aca caa ggc tac  
                 
                     
                 
                   ttc cct gat tgg cag aac tac aca cca gga cca ggg 
                 
                     
                 
                   gtt aga tat cca ctg acc ttt gga tgg tgc tac aag  
                 
                     
                 
                   cta gta cca gtt gag cca gag aag tta gaa gaa gcc 
                 
                     
                 
                   aac aaa gga gag aac acc agc ttg tta cac cct gtg  
                 
                     
                 
                   agc ctg cat gga atg gat gac ccg gcg aga gaa gtg 
                 
                     
                 
                   tta gag tgg agg ttt gac agc cgc cta gca ttt cat  
                 
                     
                 
                   cac gtg gcc cga gag ctg cat ccg gag tac ttc aag 
                 
                     
                 
                   aac tgc tga 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . Nucleotide sequence or DNA expression vector according to  claim 1 , for use according to  claim 1 , for the prevention and treatment of diseases chosen from the group consisting of chronic infective diseases such as HBV, HCV and HIV, tuberculosis and malaria, acute infective diseases such as influenza, West Nile, Crimean-Congo hemorrhagic fever and Ebola diseases, tumors such as breast, pulmonary, prostate or bladder tumor. 
     
     
         6 . Pharmaceutical composition, that is immunogenic, comprising or consisting of a nucleotide sequence expressing a fusion protein, said fusion protein comprising or consisting of the exosome-anchoring protein of sequences SEQ ID NO:1 fused at its C-terminus with an antigen, or a DNA expression vector comprising said nucleotide sequence, in association with one or more pharmaceutically acceptable excipients and/or adjuvants, wherein SEQ ID NO:1 is the following sequence: 
       
         
           
                 
                 
               
                     
                   MGCKWSKSSV VGWPAVRERM RRAEPAADGV GAASRDLEKH 
                 
                     
                     
                 
                     
                   GAITSSNTAA TNADCAWLEA QEEEEVGFPV TPQVPLRPMT  
                 
                     
                     
                 
                     
                   YKAAVDLSHF LKEKGGLEGL IHSQRRQDIL DLWIYHTQGY 
                 
                     
                     
                 
                     
                   FPDWQNYPTG PGIRYPLTFG WCYKLVPVEP EKLEEANKGE 
                 
                     
                     
                 
                     
                   NTSLLHPVSL HGMDDPGREV LEWRFDSRLA FHHVARELHP 
                 
                     
                     
                 
                     
                   EYFKNC. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         7 . Pharmaceutical composition according to  claim 6 , wherein the antigen is chosen from the group consisting of Human Papilloma virus antigen such as E6 and E7, HIV antigen such as Gag and Tat, Ebola virus antigen such as VP24, VP40, NP and GP, West Nile virus antigen such as NS3, HBV antigen such as Core, HCB antigen such as Core, NS3, E1 and E2; Crimean-Congo virus antigen such as NP and GP; Influenza A virus antigen such as NP and M1; human melanoma antigen such as MAGE-A3 and MART-1; human tumor-associated antigens such as Her2/Neu, HoxB7. 
     
     
         8 . Pharmaceutical composition according to  claim 6 , wherein the nucleotide sequence expressing the exosome-anchoring protein of sequence SEQ ID NO:1 is the following sequence SEQ ID NO:2: 
       
         
           
                 
               
                   atg ggt tgc aag tgg tca aaa agt agt gtg gtt gga  
                 
                     
                 
                   tgg cct gct gta agg gaa aga atg aga cga gct gag  
                 
                     
                 
                   cca gca gca gat ggg gtg gga gca gca tct cga gac 
                 
                     
                 
                   cta gaa aaa cat gga gca atc aca agt agc aat aca  
                 
                     
                 
                   gca gct acc aat gct gat tgt gcc tgg cta gaa gca 
                 
                     
                 
                   caa gag gag gag gag gtg ggt ttt cca gtc aca cct  
                 
                     
                 
                   cag gta cct tta aga cca atg act tac aag gca gct 
                 
                     
                 
                   gta gat ctt agc cac ttt tta aaa gaa aag ggg gga  
                 
                     
                 
                   ctg gaa ggg cta att cac tcc caa cga aga caa gat 
                 
                     
                 
                   atc ctt gat ctg tgg atc tac cac aca caa ggc tac  
                 
                     
                 
                   ttc cct gat tgg cag aac tac aca cca gga cca ggg 
                 
                     
                 
                   gtt aga tat cca ctg acc ttt gga tgg tgc tac aag  
                 
                     
                 
                   cta gta cca gtt gag cca gag aag tta gaa gaa gcc 
                 
                     
                 
                   aac aaa gga gag aac acc agc ttg tta cac cct gtg  
                 
                     
                 
                   agc ctg cat gga atg gat gac ccg gcg aga gaa gtg 
                 
                     
                 
                   tta gag tgg agg ttt gac agc cgc cta gca ttt cat  
                 
                     
                 
                   cac gtg gcc cga gag ctg cat ccg gag tac ttc aag 
                 
                     
                 
                   aac tgc tga 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         9 . Pharmaceutical composition according to  claim 6 , wherein the pharmaceutical composition is in a form suitable for a route of intramuscular administration. 
     
     
         10 . Pharmaceutical composition according to  claim 6 , wherein the adjuvant is an adjuvant of CD8 +  T cells response. 
     
     
         11 . Pharmaceutical composition according to  claim 6 , for use in vaccine prevention and therapy. 
     
     
         12 . Pharmaceutical composition according to  claim 11 , for use according to  claim 11 , for the prevention and treatment of diseases chosen from the group consisting of chronic infective diseases such as HBV, HCV and HIV, tuberculosis and malaria, acute infective diseases such as influenza, West Nile, Crimean-Congo hemorrhagic fever and Ebola diseases, tumors such as breast, pulmonary, prostate or bladder tumor.