Novel Isolated And Purified Strains Of Chikungunya Virus And Polynucleotides And Polypetide Sequences, Diagnostic And Immunogenical Uses Thereof

Abstract

The present invention concerns wild-strains of Chikungunya virus isolated from patients exhibiting severe forms of infection and stemming from a human arbovirosis epidemy. The present invention also concerns polypeptide sequences and fragment thereof derived from their genome, the polynucleotide encoding same and their use as diagnostic products, as vaccine and/or as immunogenic compositions.

1 . An isolated and purified wild strain of chikungunya virus (CHIKV) capable of in vitro infecting human cells. 
     
     
         2 . The strain according to  claim 1 , selected from the group consisting of the isolates 05.115, 05.61, 05.209, 06.21, 06.27 and 06.49. 
     
     
         3 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 4  (SEQ ID NO:1). 
     
     
         4 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 5  (SEQ ID NO:2). 
     
     
         5 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 6  (SEQ ID NO:3). 
     
     
         6 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 7  (SEQ ID NO:4). 
     
     
         7 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 8  (SEQ ID NO:5). 
     
     
         8 . The strain according to  claim 1  or  2 , wherein its genome comprises a sequence as shown in  FIG. 9  (SEQ ID NO:6). 
     
     
         9 . An isolated and purified strain of chikungunya virus (CHIKV) comprising at least one mutation in structural protein E1 and/or structural protein E2. 
     
     
         10 . The strain of  claim 9 , wherein the at least one mutation is located in the ectodomain of protein E 1 and/or E2. 
     
     
         11 . The strain of  claim 9  or  10 , wherein the at least one mutation in the E2 protein is a mutation at a position homologous to amino acid position 382, 399, 404, 485, 489, 506, 536, 624, 637, 669, 700 or 711 of SEQ ID NO: 23. 
     
     
         12 . The strain of  claim 11 , wherein the mutation is G382K. 
     
     
         13 . The strain of  claim 11 , wherein the mutation is 1399M. 
     
     
         14 . The strain of  claim 11 , wherein the mutation is G404E. 
     
     
         15 . The strain of  claim 11 , wherein the mutation is N495T. 
     
     
         16 . The strain of  claim 11 , wherein the mutation is A489T. 
     
     
         17 . The strain of  claim 11 , wherein the mutation is L506M. 
     
     
         18 . The strain of  claim 11 , wherein the mutation is 1536T. 
     
     
         19 . The strain of  claim 11 , wherein the mutation is S624N. 
     
     
         20 . The strain of  claim 11 , wherein the mutation is T637M. 
     
     
         21 . The strain of  claim 11 , wherein the mutation is A669T. 
     
     
         22 . The strain of  claim 11 , wherein the mutation is S700T. 
     
     
         23 . The strain of  claim 11 , wherein the mutation is V711A. 
     
     
         24 . The strain of  claim 9  or  10 , wherein the at least one mutation in the E1 protein is a mutation at a position homologous to amino acid position 1035, 1078, 1093 or 1131 of SEQ ID NO: 23. 
     
     
         25 . The strain of  claim 24 , wherein the mutation is A1035V. 
     
     
         26 . The strain of  claim 24 , wherein the mutation is M1078V. 
     
     
         27 . The strain of  claim 24 , wherein the mutation is D1093E. 
     
     
         28 . The strain of  claim 24 , wherein mutation is V1131A. 
     
     
         29 . An isolated and purified polynucleotide comprising all or part of the sequence as shown in  FIG. 4 ,  5 ,  6 ,  7 ,  8  or  9  (SEQ ID NOS 1, 2, 3, 4, 5 or 6). 
     
     
         30 . Fragment of the polynucleotide as defined in  claim 29 , wherein it codes for the ectodomain of glycoprotein E2. 
     
     
         31 . The fragment of  claim 30 , wherein it comprises a nucleotide sequence as shown in  FIG. 10 ,  11 ,  12  or  13  (SEQ 10 NOS 7, 8, 9 or 10). 
     
     
         32 . Fragment of the polynucleotide as defined in  claim 29 , wherein it codes for a soluble form of glycoprotein E2. 
     
     
         33 . The fragment of  claim 32 , wherein it consists of a nucleotide sequence as shown in  FIG. 14 ,  15 ,  16  or  17  (SEQ ID NOS 11, 12, 13 or 14). 
     
     
         34 . The fragment of the polynucleotide as defined in  claim 29 , wherein it codes for the ectodomain of glycoprotein E1. 
     
     
         35 . A vector comprising a fragment as defined in any one of  claims 30  to  34 . 
     
     
         36 . A plasmid comprising a fragment as defined in  claim 31 , selected from the group deposited at the CNCM (Collection Nationale de Cultures de Microorganismes), 28 rue du Docteur Roux, 75724 PARIS Cedex 15, France, on Mar. 15, 2006 under accession number 1-3587, 1-3588, 1-3589 and 1-3590. 
     
     
         37 . A plasmid comprising a fragment as defined in  claim 33 , wherein the sequence in optimised for efficient production of recombinant E2 protein deposited at the CNCM (Collection Nationale de Cultures de Microorganismes), 28 rue du Oocteur Raux, 75724 PARIS Cedex 15, France, on Mar. 14, 2007, under accession number 1-3733. 
     
     
         38 . A host cell comprising a vector as defined in  claim 35 . 
     
     
         39 . A host cell comprising a plasmid as defined in  claim 36  or  37 . 
     
     
         40 . A purified polypeptide encoded by a polynucleotide as defined in  claim 29 , or by a fragment as defined in anyone of  claims 30  to  34 . 
     
     
         41 . A purified polypeptide comprising all or part of the sequence as defined in  FIGS. 38 to 49  (SEQ ID NOS: 24 to 34 and 78) 
     
     
         42 . A purified polypeptide comprising all or part of the sequence as defined in  FIG. 18 ,  19 ,  20  or  21  (SEQ ID NOS 15, 16, 17, or 18). 
     
     
         43 . A purified polypeptide comprising all or part of the sequence as defined in  FIG. 22 ,  23 ,  24  or  25  (SEQ ID NOS 19, 20, 21, or 22). 
     
     
         44 . Monoclonal or polyclonal antibodies, or fragment thereof, that specifically bind to a polypeptide as defined in anyone of  claims 40  to  43 . 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . A composition comprising at least one element selected from the group consisting of:
 a strain as defined in anyone of  claims 1  to  28 ;   a polynucleotide as defined in  claim 29 ;   a fragment as defined in anyone of  claims 30  to  34 ;   a vector as defined in anyone of  claims 35  to  37 ;   a plasmid as defined in  claim 36  or  37 ;   a host cell as defined in  claim 38  or  39 ;   a polypeptide as defined in anyone of  claims 40  to  43 ; and   an antibody as defined in  claim 44 ;   
       for treating and/or preventing an arbovirosis. 
     
     
         48 . (canceled) 
     
     
         49 . A kit for the detection of a CHIKV associated to an arbovirosis, comprising at least one element selected from the group consisting of:
 a strain as defined in anyone of  claims 1  to  28 ;   a polynucleotide as defined in  claim 29 ;   a fragment as defined in anyone of  claims 30  to  34 ;   a vector as defined in anyone of  claims 35  to  37 ;   a plasmid as defined in  claim 36  or  37 ;   a host cell as defined in  claim 38  or  39 ;   a polypeptide as defined in anyone of  claims 40  to  43 ; and   an antibody as defined in  claim 44 .