Fusion protein of hiv regulatory/accessory proteins

Abstract

The invention relates to fusion proteins comprising the amino acid sequence of at least four HIV proteins selected from Vif, Vpr, Vpu, Rev, Tat and Nef or derivatives of the amino acid sequence of one or more of said proteins, wherein the fusion protein is not processed to individual HIV proteins having the natural N and C termini. The invention further concerns nucleic acids encoding said proteins, vectors comprising said nucleic acids, and methods for producing said proteins. The fusion protein, nucleic acids and vectors are usable as vaccines for the at least partial prophylaxis against HIV infections.

1 . A fusion protein comprising the amino acid sequence of at least four HIV proteins selected from Vif, Vpr, Vpu, Vpx, Rev, Tat and Nef or derivatives of the amino acid sequence of one or more of said proteins, wherein the fusion protein is not processed to individual HIV proteins having the natural N and C termini and wherein a derivative of the amino acid sequence of an HIV protein is an amino acid sequence showing a homology of at least 50%, when the corresponding part of the amino acid sequence in the fusion protein is compared to the amino acid sequence of the respective HIV protein of known HIV isolates.  
   
   
       2 . Fusion protein according to  claim 1 , wherein the homology is at least 80%.  
   
   
       3 . Fusion protein according to  claim 1 , wherein not more than 10 amino acids are deleted, inserted or substituted in the derivative of the amino acid sequence when compared to the amino acid sequence of the respective HIV protein of known HIV isolates to obtain an HIV protein with reduced activity or no activity at all.  
   
   
       4 . Fusion protein according to anyone of  claims 1  to  3 , wherein the HIV proteins are selected from Vif, Vpr, Vpx, Vpu, Rev and Tat  
   
   
       5 . Fusion protein according to anyone of  claims 1  to  4 , comprising the amino acid sequence of the HIV proteins Vif, Vpr, Vpu, Rev and Tat or derivatives of the amino acid sequence of one or more of said proteins.  
   
   
       6 . Fusion protein according to anyone of  claims 1  to  5 , wherein the amino acid sequences of at least two of the HIV proteins are fused to each other without additional amino acids.  
   
   
       7 . Fusion protein according to anyone of  claims 1  to  6 , wherein the amino acid sequences of at least two of the HIV proteins are separated by at least one additional amino acid.  
   
   
       8 . Fusion protein according to anyone of  claims 1  to  7 , wherein the amino acid sequence of at least one of the HIV proteins is fused to a fusion partner which is not a HIV protein selected from Vif, Vpr, Vpx, Vpu, Rev, Tat and Nef.  
   
   
       9 . Nucleic acid encoding a fusion protein according to anyone of  claims 1  to  8 .  
   
   
       10 . Nucleic acid according to  claim 9 , wherein the nucleic acid is DNA.  
   
   
       11 . Nucleic acid according to  claim 10 , wherein the expression of the fusion protein from the DNA is controlled by regulatory elements selected from eukaryotic, procaryotic and viral promoters.  
   
   
       12 . Nucleic acid according to  claim 11 , wherein the viral promoter is a poxyiral promoter.  
   
   
       13 . Nucleic acid according to anyone of  claims 9  to  12 , wherein the nucleic acid further comprises the coding sequence for at least one additional HIV protein selected from Gag, Pol and Env.  
   
   
       14 . Nucleic acid according to  claim 13 , wherein the nucleic acid comprises the coding sequence for the HIV Gag, Pol and Env proteins.  
   
   
       15 . Vector comprising a nucleic acid according to anyone of  claims 9  to  14 .  
   
   
       16 . Vector according to claim anyone of claims  15 , wherein the vector is a viral vector.  
   
   
       17 . Vector according to  claim 16 , wherein the viral vector is a poxvirus vector, in particular a Vaccinia Virus vector.  
   
   
       18 . Vector according to  claim 17 , wherein the Vaccinia virus vector is Modified Vaccinia Virus Ankara (MVA).  
   
   
       19 . Vector according to  claim 18 , wherein MVA is selected from MVA-575 deposited at the European Collection of Animal Cell Cultures (ECACC) under the deposition number V00120707 and MVA-BN deposited at the ECACC under the deposition number V00083008.  
   
   
       20 . Method of producing a protein according to anyone of  claims 1  to  8 , comprising the steps of 
 transfecting a host cell with a nucleic acid according to anyone of  claims 9  to  14  or a with a vector according  claim 15  or    infecting a host cell with a viral vector according to anyone of  claims 16  to  19 ,    expressing the fusion protein in the transfected host cell or the infected host cell, and    recovering the fusion protein.    
   
   
       21 . Host cell transfected with a nucleic acid according to anyone of  claims 9  to  14  or a vector according to  claim 15  or infected with a viral vector according to anyone of  claims 16  to  19 .  
   
   
       22 . Fusion protein according to anyone of  claims 1  to  8 , nucleic acid according to anyone of  claims 9  to  14  or vector according to anyone of  claims 15  to  19  as a medicament.  
   
   
       23 . Fusion protein according to anyone of  claims 1  to  8 , nucleic acid according to anyone of  claims 9  to  14  or vector according to anyone of  claims 15  to  19  as a vaccine.  
   
   
       24 . Vaccine comprising a fusion protein according to anyone of  claims 1  to  8 , a nucleic acid according to anyone of  claims 9  to  14  or a vector according to anyone of  claims 15  to  19 .  
   
   
       25 . Use of a fusion protein according to anyone of  claims 1  to  8 , of a nucleic acid according to anyone of  claims 9  to  14  or of a vector according to anyone of  claims 15  to  19  for the preparation of a vaccine.  
   
   
       26 . Method for protecting an animal, including a human, against an HIV infection by administering to an animal, including a human, in need thereof a fusion protein according to anyone of  claims 1  to  8 , a nucleic acid according to anyone of  claims 9  to  14  or a vector according to anyone of  claims 15  to  19 .