Recombinant measles viruses expressing epitopes of antigens of RNA viruses - use for the preparation of vaccine compositions

Abstract

The invention relates to a recombinant measles virus expressing a heterologous amino acid sequence derived from an antigen of a determined RNA virus, said recombinant measles virus being capable of eliciting a humoral and/or cellular immune response against measles virus or against said RNA virus or against both measles virus and against said RNA virus. It also relates to the use of said recombinant measles virus for the preparation of immunogenic composition.

1 . Recombinant mononegavirales virus expressing a heterologous amino acid sequence, said recombinant virus being capable of eliciting a humoral and/or a cellular immune response against said heterologous amino acid sequence including in individuals having pre-existing measles virus immunity.  
     
     
         2 . Recombinant measles virus expressing a heterologous amino acid sequence derived from an antigen of a determined RNA virus, said recombinant measles virus being capable of eliciting a humoral and/or a cellular immune response against measles virus or against said RNA virus or against both measles virus and against said RNA virus.  
     
     
         3 . Recombinant measles virus according to  claim 2  which is the product of the expression in a cell of a recombinant nucleotide sequence comprising acDNA molecule encoding the full length antigenomic (+) RNA of the measles virus (MV) and further comprising, recombined with said cDNA molecule, a sequence encoding an heterologous amino acid sequence of a determined retrovirus or flavivirus.  
     
     
         4 . Recombinant measles virus according to claims  1  or  2 , which is derived from the Schwarz vaccine strain or from the Edmonston strain.  
     
     
         5 . Recombinant measles virus according to anyone of  claims 2  to  4 , which is rescued from helper cells transfected with a recombinant nucleotide sequence which comprises cDNA encoding the nucleotid sequence of full length antigenomic (+) RNA of the measles virus, said cDNA being recombined with a nucleotide sequence encoding a retroviral orflaviviral heterologous amino acid sequence, and said recombinant nucleotide sequence complying with the rule of six.  
     
     
         6 . Recombinant measles virus according to anyone of  claims 2  to  5 , wherein the nucleotide sequence is comprised within the EdB-tag virus vector recombined with the ATU inserted in a position of the EdB-tag vector taking advantage of the gradient of the viral genome to allow various levels of expression of the transgenic nucleotide sequence encoding the heterologous amino acid sequence inserted in said ATU.  
     
     
         7 . Recombinant measles virus according to anyone of  claims 2  to  5 , which is the product of the expression of a recombinant nucleotide sequence comprising a cDNA molecule which encodes the nucleotide sequence of the full length antigenomic (+) RNA of a measles virus (MV) originating from an approved vaccine strain, wherein said cDNA molecule is recombined with a heterologous nucleotide sequence encoding a heterologous amino acid sequence derived from an antigen of a determined heterologous retrovirus or flavivirus.  
     
     
         8 . Recombinant measles virus according to  claim 7 , wherein the cDNA molecule comprises the insert contained in plasmid pTM-MVSchw deposited on Jun. 12, 2002 under No. 1-2889, wherein said insert encodes the nucleotide sequence of the full length antigenomic (+) RNA strand of the measles virus.  
     
     
         9 . Recombinant measles virus according to  claim 7  or  8 , wherein the recombinant nucleotide sequence is derived from the insert contained in plasmid pTM-MVSchw2-gfp deposited on Jun. 12, 2002 under I-2890 (CNCM), wherein the sequence of the gfp gene is substituted for a sequence encoding a determined amino acid sequence.  
     
     
         10 . Recombinant measles virus according to anyone of  claims 7  to  9 , wherein the cDNA molecule is selected among the following sequences: 
 nucleotide sequence extending from nucleotide 83 to nucleotide 15977 of  FIG. 11     nucleotide sequence extending from nucleotide 29 to nucleotide 15977 of  FIG. 11     nucleotide sequence extending from nucleotide 29 to nucleotide 16202 of  FIG. 11     nucleotide sequence extending from nucleotide 26 to nucleotide 15977 of  FIG. 11     nucleotide sequence extending from nucleotide 26 to nucleotide 16202 of  FIG. 11     nucleotide sequence extending from nucleotide 9 to nucleotide 15977 of  FIG. 11     nucleotide sequence extending from nucleotide 9 to nucleotide 16202 of  FIG. 11     
     
     
         11 . Recombinant measles virus according to anyone of  claims 2  to  10 , wherein the heterologous amino acid sequence is derived from an antigen of a retrovirus selected among HIV retroviruses, or from an antigen of a flavivirus.  
     
     
         12 . Recombinant measles virus according to anyone of  claims 2  to  9 , wherein the heterologous amino acid sequence is derived from an antigen of the Yellow Fever Virus or the West Nile Virus.  
     
     
         13 . Recombinant measles virus according to anyone of  claims 2  to  11 , wherein the heterologous amino acid sequence is derived from an envelope antigen of the HIV retrovirus.  
     
     
         14 . Recombinant measles virus according to  claim 13 , wherein the heterologous amino acid sequence is a recombinant gp160 or a recombinant gp120 antigens of HIV-1.  
     
     
         15 . Recombinant measles virus according to  claim 14 , wherein the V1, V2 and/or V3 loops of the gp120 antigen are deleted or deleted in part, individually or in combination in such a way that conserved epitopes are exposed on the obtained recombinant gp120 antigen.  
     
     
         16 . Recombinant measles virus according to  claim 15 , wherein the V1, V2 and/or V3 loops of the gp120 antigen are substituted or substituted in part, individually or in combination in such a way that conserved epitopes are exposed on the obtained recombinant gp120 antigen.  
     
     
         17 . Recombinant measles virus according to  claim 16 , wherein the V3 loop is substituted for a gp41 epitope such as AAELDKWASAA (SEQ ID NO: 8).  
     
     
         18 . Recombinant measles virus according to anyone of  claims 11  to  16 , wherein the heterologous amino acid sequence isgp160 ΔV3, gp160 ΔV1V2, gp160 ΔV1V2V3, gp140 ΔV3, gp140 ΔV1V2, gp140 ΔV1V2V3.  
     
     
         19 . Recombinant measles virus according to anyone of  claims 2  to  12 , wherein the amino acid sequence is derived from an antigen of the Yellow Fever virus selected among the envelope (Env) or the NS1 proteins or immunogenic mutants thereof.  
     
     
         20 . Recombinant measles virus according to anyone of  claims 2  to  12 , wherein the amino acid sequence is derived from an antigen of the West Nile virus selected among the envelope (E), premembrane(preM) or immunogenic mutants thereof.  
     
     
         21 . Recombinant measles virus according to anyone of  claims 2  to  12 , capable of inducing protection against heterologous VLP or pM/E proteins.  
     
     
         22 . Recombinant measles virus according to anyone of  claims 2  to  21 , which elicits a humoral and/or a cellular immune response in an animal model susceptible to measles virus.  
     
     
         23 . Recombinant measles virus according to anyone of  claims 2  to  22 , which elicits neutralizing antibodies against the heterologous amino acid sequence in a mammalian animal model susceptible to measles virus.  
     
     
         24 . Recombinant measles virus according to anyone of  claims 2  to  23  wherein the heterologous amino acid sequence is derived from an envelope proteins of HIV-1 and which elicits antibodies capable of neutralizing a primary HIV isolate when tested on indicator cells such as P4-CCR5 cells.  
     
     
         25 . Recombinant measles virus according to  claims 2  to  24  which elicits neutralizing antibodies against the heterologous amino acid sequence in a mammal.  
     
     
         26 . Recombinant measles virus vector comprising a replicon comprising (i) a cDNA sequence encoding the full length antigenomic (+) RNA of a measles virus operatively linked to (ii) expression control sequences and (iii) a heterologous DNA sequence coding for a determined heterologous amino acid sequence, said heterologous DNA sequence being cloned in said replicon in conditions allowing its expression and said replicon having a total number of nucleotides which complies with the rule of six.  
     
     
         27 . Recombinant measles virus vector according to  claim 26 , wherein the heterologous DNA sequence is cloned within an Additional Transcription Unit (ATU) inserted in the cDNA corresponding to the antigenomic RNA of measles virus.  
     
     
         28 . Recombinant measles virus vector according to  claim 26 , wherein the cloning site of the ATU is chosen upstream from the N gene of the MV virus.  
     
     
         29 . Recombinant measles virus vector according to  claim 26 , wherein the cloning site of the ATU is chosen between the P and M genes of the MV virus.  
     
     
         30 . Recombinant measles virus vector according to  claim 26 , wherein the cloning site of the ATU is between the H and L genes of the MV virus.  
     
     
         31 . Recombinant measles virus vector according to anyone of  claims 26  to  29 , wherein the heterologous DNA sequence is expressed as a fusion protein with one of the MV proteins.  
     
     
         32 . Recombinant measles virus vector according to anyone of  claims 26  to  29 , wherein the heterologous DNA sequence is not expressed as a fusion protein with one of the MV proteins.  
     
     
         33 . Recombinant measles virus vector according to anyone of  claims 26  to  29  wherein the heterologous DNA sequence encodes a retroviral amino acid sequence.  
     
     
         34 . Recombinant measles virus vector according to anyone of  claims 26  to  32  wherein the heterologous DNA sequence encodes a retroviral amino acid sequence derived from an antigen of a retrovirus selected among HIV retroviruses.  
     
     
         35 . Recombinant measles virus vector according to anyone of  claims 26  to  32  wherein the heterologous DNA sequence encodes a retroviral amino acid sequence derived from an antigen of a retrovirus selected among flaviviruses.  
     
     
         36 . Recombinant measles virus vector according to anyone of  claims 26  to  33  wherein the heterologous DNA sequence encodes a retroviral amino acid sequence derived from an envelope antigen of an HIV retrovirus.  
     
     
         37 . Recombinant measles virus vector according to anyone of  claims 26  to  33  wherein the heterologous DNA sequence encodes a retroviral amino acid sequence selected among the gp160, the gp120 orgp41 of HIV-1, or the gp140 of HIV-1, or a mutated version of said antigens.  
     
     
         38 . Recombinant measles virus vector according to anyone of  claims 26  to  36 , wherein the mutated antigen enables exposition of neutralizing epitopes.  
     
     
         39 . Recombinant measles virus vector according to anyone of  claims 26  to  37 , wherein the heterologous DNA sequence encodes gp160AV3, gp160 ΔV1V2, gp160 ΔV1V2V3, gp140 ΔV3, gp140 ΔV1V2, gp140 ΔV1V2V3.  
     
     
         40 . Recombinant measles virus vector according to anyone of  claims 25  to  32  wherein the heterologous DNA sequence encodes a flaviviral amino acid sequence derived from an antigen of the Yellow Fever virus or the West Nile virus.  
     
     
         41 . Recombinant measles virus vector according to anyone of  claims 26  to  40  wherein the nucleotide sequence comprising cDNA resulting from reverse transcription of the antigenic RNA of measles virus, originates from a measles virus strain approved for vaccination.  
     
     
         42 . Recombinant measles virus vector according to  claim 41 , wherein the measles virus strain is the Schwarz strain.  
     
     
         43 . Recombinant measles virus vector according to  claim 42 , wherein the cDNA encoding the full length antigenomic (+) RNA of the measles virus and the expression control sequence are derived from pTM-MVSchw deposited at the CNCM under No. 1-2889.  
     
     
         44 . Recombinant measles virus vector according to anyone of  claims 26  to  42 , which is a plasmid.  
     
     
         45 . Recombinant measles virus vector according to anyone of  claims 26  to  44  wherein the replicon is designed according to the map of  FIG. 2  wherein insert represents the heterologous DNA sequence.  
     
     
         46 . Recombinant measles virus vector according to anyone of  claims 26  to  45 , wherein the heterologous DNA sequence is selected among YFV17D204.  
     
     
         47 . Recombinant measles virus vector according to anyone of  claims 26  to  45 , wherein the heterologous DNA sequence is selected among the neurovirulent strain IS-98-ST1.  
     
     
         48 . Recombinant measles virus vector according to  claim 43 , which is selected among the following vectors deposited with the CNCM  
       
         
           
                 
                 
               
                     
                 
                     
                 
                   pMV2 (EdB)gp160 [delta] V3HIV89. 6P 
                   CNCM 1-2883 
                 
                   pMV2 (EdB) gp160HIV89. 6P 
                   CNCM 1-2884 
                 
                   pMV2 (EdB) gp140HIV89. 6P 
                   CNCM 1-2885 
                 
                   pMV3 (EdB)gp140 [delta] V3HIV89. 6P 
                   CNCM 1-2886 
                 
                   pMV2(EdB)-NS1YFV17D 
                   CNCM I-2887 
                 
                   pMV2 (EdB)-EnvYFV17D 
                   CNCM 1-2888. 
                 
                   pTM-MVSchw2-Es(WNV) 
                   CNCM I-3033 
                 
                   pTM-MVSchw2-GFPbis- 
                   CNCM 1-3034 
                 
                   pTM-MVSchw2-p17p24[delta] myr(HIVB) 
                   CNCM 1-3035 
                 
                   pTM-MVSchw3-Tat(HIV89-6p) 
                   CNCM I-3036 
                 
                   pTM-MVschw3-GFP 
                   CNCM 1-3037 
                 
                   pTM-MVSchw2-Es (YFV) 
                   CNCM 1-3038 
                 
                   pTM-MVSchw2-gp140 [delta] V1 V2 V3(HIV89-6) 
                   CNCM 1-3054 
                 
                   pTM-MVSchw2-gp140 [delta] V3(HIV89-6) 
                   CNMC I-3055 
                 
                   pTM-MVSchw2-gp160 [dela] V1 V2 V3(HIV89-6) 
                   CNCM 1-3056 
                 
                   pTM-MVSchw2-gp160 [delta] V1 V2(HIV89-6) 
                   CNCM 1-3057 
                 
                   pTM-MVSchw2-Gag SIV239 p17-p24 [delta] myr-3-gp 
                   CNCM I-3058. 
                 
                   140 (HIV89-6) 
                 
                     
                 
                     
                 
             
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         49 . A rescue system for the assembly of recombinant measles virus expressing a heterologous amino acid sequence, which comprises a determined cell transfected with a recombinant measles virus vector according to anyone of  claims 26  to  47 , and a determined helper cell recombined with at least one vector suitable for expression of T7 RNA polymerase and expression of the N, P and L proteins of the measles virus.  
     
     
         50 . An immunogenic composition comprising a recombinant virus according to anyone of  claims 1  to  26 , or a recombinant vector according to anyone of claims to  48 .  
     
     
         51 . A vaccine composition comprising a recombinant virus according to anyone of  claims 1  to  26 , or a recombinant vector according to anyone of claims to  48 .