Use of proteins and peptides encoded by the genome of a novel sars-associated coronavirus strain

Abstract

The invention relates to the use of proteins and peptides coded by the genome of the isolated or purified strain of severe acute respiratory syndrome (SARS)-associated coronavirus, resulting from sample reference number 031589 and, in particular, to the use of protein S and the derivative antibodies thereof as diagnostic reagents and as a vaccine.

1 . An isolated and purified protein or polypeptide, characterized in that it is the S protein having the sequence SEQ ID No: 3, its ectodomain or a fragment of its ectodomain. 
     
     
         2 . The protein or polypeptide as claimed in  claim 1 , characterized in that it consists of the amino acids corresponding to positions 1 to 1193 of the amino acid sequence of the S protein. 
     
     
         3 . The protein or polypeptide as claimed in  claim 1 , characterized in that it consists of the amino acids corresponding to positions 14 to 1193 of the amino acid sequence of the S protein. 
     
     
         4 . The isolated protein or polypeptide as claimed in  claim 1 , characterized in that it consists of the amino acids corresponding to positions 475 to 1193 of the amino acid sequence of the S protein. 
     
     
         5 . A nucleic acid encoding a protein or a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         6 . The nucleic acid as claimed in  claim 5 , characterized in that it comprises the sequence encoding SEQ ID No: 5 or the sequence encoding SEQ ID No: 6. 
     
     
         7 . A recombinant expression vector, characterized in that it encodes a protein or a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         8 . The recombinant expression vector as claimed in  claim 7 , characterized in that it is chosen from the vectors contained in the following bacterial strains, deposited at the Collection Nationale de Cultures de Microorganismes (CNCM), 25 rue du Docteur Roux, 75724 Paris Cedex 15:
 a) strain No. I-3118, deposited on Oct. 23, 2003,   b) strain No. I-3019, deposited on May 12, 2003,   c) strain No. I-3020, deposited on May 12, 2003,   d) strain No. I-3059, deposited on Jun. 20, 2003,   e) strain No. I-3323, deposited on Nov. 22, 2004,   f) strain No. I-3324, deposited on Nov. 22, 2004,   g) strain No. I-3326, deposited on Dec. 1, 2004,   h) strain No. I-3327, deposited on Dec. 1, 2004,   i) strain No. I-3332, deposited on Dec. 1, 2004,   j) strain No. I-3333, deposited on Dec. 1, 2004,   k) strain No. I-3334, deposited on Dec. 1, 2004,   l) strain No. I-3335, deposited on Dec. 1, 2004,   m) strain No. I-3336, deposited on Dec. 1, 2004,   n) strain No. I-3337, deposited on Dec. 1, 2004,   o) strain No. I-3338, deposited on Dec. 2, 2004,   p) strain No. I-3339, deposited on Dec. 2, 2004,   q) strain No. I-3340, deposited on Dec. 2, 2004, and   r) strain No. I-3341, deposited on Dec. 2, 2004.   
     
     
         9 . A nucleic acid containing a synthetic gene allowing optimized expression of the S protein in eukaryotic cells, characterized in that it possesses the sequence SEQ ID No: 140. 
     
     
         10 . An expression vector containing a nucleic acid as claimed in  claim 9 , characterized in that it is contained in the bacterial strain deposited at the CNCM, on Dec. 1, 2004, under the No. I-3333. 
     
     
         11 . The expression vector as claimed in  claim 7  or  claim 9 , characterized in that it is a viral vector, in the form of a viral particle or in the form of a recombinant genome. 
     
     
         12 . The vector as claimed in  claim 11 , characterized in that it is a recombinant viral particle or a recombinant viral genome capable of being obtained by transfecting a plasmid according to paragraphs g), h) or k) to r) of  claim 8 , into an appropriate cellular system. 
     
     
         13 . A lentiviral vector encoding a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         14 . A recombinant measles virus encoding a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         15 . A recombinant vaccinia virus encoding a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         16 . The use of a vector according to paragraphs d) to p) of  claim 8 , or of a vector as claimed in  claim 10 , for the production, in a eukaryotic system, of the SARS-associated coronavirus S protein or of a fragment of this protein. 
     
     
         17 . A method for producing the S protein in a eukaryotic system, comprising a step of transfecting eukaryotic cells in culture with a vector chosen from the vectors contained in the bacterial strains mentioned in paragraphs d) to p) of  claim 8 , or in  claim 10 . 
     
     
         18 . A genetically modified eukaryotic cell expressing a protein or a polypeptide as claimed in any one of  claims 1  to  4 . 
     
     
         19 . The cell as claimed in  claim 18 , capable of being obtained by transfection with any one of the vectors mentioned in paragraphs k) to n) of  claim 8 . 
     
     
         20 . The cell as claimed in  claim 19 , characterized in that it is the cell FRhK4-Ssol-30, deposited at the CNCM on Nov. 22, 2004, under the No. I-3325. 
     
     
         21 . A monoclonal antibody recognizing the native S protein of a SARS-associated coronavirus. 
     
     
         22 . The use of a protein or a polypeptide as claimed in any one of  claims 1  to  4 , or of an antibody as claimed in  claim 21 , for detecting a SARS-associated coronavirus infection, from a biological sample. 
     
     
         23 . A method for detecting a SARS-associated coronavirus, from a biological sample, characterized in that the detection is carried out by ELISA using the recombinant S protein or its ectodomain, or a fragment of its ectodomain, expressed in a eukaryotic system. 
     
     
         24 . The method of detection as claimed in  claim 23 , additionally comprising a step of detection by ELISA using the recombinant N protein. 
     
     
         25 . The method as claimed in  claim 23  or  24 , characterized in that it is a double epitope ELISA method, and in that the serum to be tested is mixed with the visualizing antigen, said mixture then being brought into contact with the antigen attached to a solid support. 
     
     
         26 . An immune complex formed of a monoclonal antibody or antibody fragment as claimed in  claim 21 , and of a SARS-associated coronavirus protein or peptide 
     
     
         27 . An immune complex formed of a protein or a polypeptide as claimed in any one of  claims 1  to  4 , and of an antibody directed specifically against an epitope of the SARS-associated coronavirus. 
     
     
         28 . A SARS-associated coronavirus detection kit or box, characterized in that it comprises at least one reagent selected from the group consisting of: a protein or polypeptide as claimed in any one of  claims 1  to  4 , a nucleic acid as claimed in either of  claims 5  and  6 , a cell as claimed in any one of  claims 18  to  20 , or an antibody as claimed in  claim 21 . 
     
     
         29 . An immunogenic and/or vaccine composition, characterized in that it comprises a recombinant protein or polypeptide as claimed in any one of  claims 1  to  4 , obtained in a eukaryotic expression system. 
     
     
         30 . An immunogenic and/or vaccine composition, characterized in that it comprises a recombinant vector or virus as claimed in any one of  claims 7 ,  8 , and  10  to  15 . 
     
     
         31 . A nucleic acid insert of viral origin, characterized in that it is contained in any one of the strains mentioned in paragraphs a) to h) and k) to r) of  claim 8 .