US2003235835A1PendingUtilityA1

Cloned DNA sequences related to the entire genomic RNA of human immunodeficiency virus II (HIV-2), polypeptides encoded by these DNA sequences and use of these DNA clones and polypeptides in diagnostic kits

Assignee: PASTEUR INSTITUTPriority: Jan 22, 1986Filed: Nov 25, 2002Published: Dec 25, 2003
Est. expiryJan 22, 2006(expired)· nominal 20-yr term from priority
C07K 14/005C12N 2740/16222C12N 2740/16122C07K 7/06A61P 31/18G01N 2333/162C12N 2740/16322C12Q 1/703A61K 38/00G01N 33/56988G01N 2469/20C12N 2740/16021C12N 7/00A61K 39/00
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Claims

Abstract

A method for diagnosing an HIV-2 (LAV-II) infection and a kit containing reagents for the same is disclosed. These reagents include cDNA probes which are capable of hybridizing to at least a portion of the genome of HIV-2. In one embodiment, the DNA probes are capable of hybridizing to the entire genome of HIV-2. These reagents also include polypeptides encoded by some of these DNA sequences.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method for diagnosing an HIV-2 infection which comprises: 
 (a) contacting genetic DNA or RNA from a body sample obtained from a person suspected of having an HIV-2 infection with a DNA probe derived from at least a portion of the genome of the HIV-2 virus; and    (b) determining whether a hybridized complex is created.    
     
     
         2 . The method of  claim 1  wherein said body sample is selected from the group consisting of tissue, blood cells, cells and body fluids.  
     
     
         3 . The method of  claim 1  wherein the presence of the hybridized complex is determined by a process selected from the group consisting of Southern blot, Northern blot and dot blot.  
     
     
         4 . The method of  claim 1  wherein the cDNA probe is analogous to the entire genome of the HIV-2 virus.  
     
     
         5 . A DNA probe capable of hybridizing to the entire genome of the HIV-2 virus.  
     
     
         6 . A method for diagnosing an HIV-2 infection which comprises: 
 (a) contacting sera obtained from a patient suspected of having an HIV-2 infection with a polypeptide expression product of a DNA segment derived from the genome of the HIV-2 virus; and    (b) determining whether an immunocomplex is formed.    
     
     
         7 . The method of  claim 6  wherein the formation of the immunocomplex is determined by a process selected from the group consisting of radioimmunoassays (RIA), radioimmunoprecipitation assays (RIPA), immunofluoresence assays (IFA), enzyme-linked immunosorbent assays (ELISA) and Western blots.  
     
     
         8 . A process for detecting the presence of a virus selected from the group consisting of LAV-II, HIV-2, STLV-III and other viruses which form complexes with LAV-II reagents comprising: 
 (a) contacting DNA or RNA from a sample suspected of containing viral genetic material with a DNA probe derived from a portion of the genome of the HIV-2 virus; and    (b) determining whether a hybridized complex is created.    
     
     
         9 . A peptide selected from the group consisting of env1, env2, env3, env4, env5, env6, env7, env8, env9, env10, env11 and gag1.  
     
     
         10 . A kit for diagnosing an HIV-2 infection by the method of  claim 6  and comprising env1, env2, env3 and gag1 peptides as the polypeptide expression product.  
     
     
         11 . A vaccinating agent comprising at least one peptide selected from the group consisting of env4, env5, env6, env7, env8, env9, env10 and env11 in admixture with suitable carriers.  
     
     
         12 . A peptide having common immunological properties with the peptide structure of the envelope glycoprotein of a virus of the HIV-2 class, said peptide having no more than 40 amino acid residues.  
     
     
         13 . A peptide according to  claim 12  having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   XR--A-E-D-YL-DQ--L--WGC-----CZ 
                     
                 
                     
                     
                 
                     
                      XA-E-D-YL-DZ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of either of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   RVTAIEKYLQDQARLNSWGCAFRQVC 
                     
                 
                     
                     
                 
                     
                      AIEKYLQDQ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
     
     
         14 . A peptide according to  claim 12  having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   X--E--Q-QQEKN--EL--L---Z 
                     
                 
                     
                     
                 
                     
                        XQ-QQEKNZ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of either of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   SLEQAQIQQEKNIVIYELQKLNSW 
                     
                 
                     
                     
                 
                     
                        QIQQEKN 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
     
     
         15 . A peptide according to  claim 12  characterized as having either of the following formulas  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   XEL--YK-V-I-P-G--APTK-KR-----Z 
                     
                 
                     
                     
                 
                     
                       XYK-V-T-P-G-APTK-KRZ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of either of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   ELGDYKLVEITPIGFAPTKEKRYSSAH 
                     
                 
                     
                     
                 
                     
                       YKLVEITPIGFAPTKEK 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
     
     
         16 . A peptide according to  claim 12  characterized as having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   x----VTV-YGVP-WK-AT--LPCA-Z 
                     
                 
                     
                     
                 
                     
                       XVTV-YGVP-WK-ATZ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   CTQYVTVFYGVPTWKNATIPLECAT 
                     
                 
                     
                     
                 
                     
                       VTVFYGVPTWKNAT 
                 
                     
                     
                 
                     
                   EKLWVTVYYGVPVWKEATTTLFCAS 
                 
                     
                     
                 
                     
                       VTVYYGVPVWKEAT 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         17 . A peptide according to  claim 16  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   CTQYVTVFYGVPTWKNATIPLFCAT 
                     
                 
                     
                         VTVYYGVPTWKNAT 
                 
                     
                     
                 
                     
                   EKLWVTVYYGVPVWKEATTTLFCAS 
                 
                     
                         VTVYYGVPVWKEAT 
                 
                     
                     
                 
                     
                   EDLWVTVYYGVPVWKEATTTLFCAS 
                 
                     
                         VTVYYGVPVWKEAT 
                 
                     
                     
                 
                     
                   DNLWVTVYYGVPVWKEATTTLFCAS 
                 
                     
                         VTVYYGVPVWKEAT 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         18 . A peptide according to  claim 12  characterized as having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   X---QE--L-NVTE-F--W-NZ 
                     
                 
                     
                           XL-NVTE-FZ 
                 
                     
                     
                 
             
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   DDYQEITL-NVTEAFDAWNN 
                     
                 
                     
                          L-NVTE 
                 
                     
                     
                 
                     
                   PNPQEVVLVNVTENFNMWKN 
                 
                     
                          LVNVTE 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         19 . A peptide according to  claim 18  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   DDYQEITL-NVTEAFDAWNN 
                     
                 
                     
                          L-NVTEAF 
                 
                     
                     
                 
                     
                   PNPQEVVLVNVTENFNMWKN 
                 
                     
                          LVNVTENF 
                 
                     
                     
                 
                     
                   PNPQEIELENVTEGFNMWKN 
                 
                     
                          LENVTEGF 
                 
                     
                     
                 
                     
                   PNPQEIALENVTENFNMWKN 
                 
                     
                          LENVTENF 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         20 . A peptide according to  claim 12  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   XL---S-KPCVKLTPLCV--KZ 
                     
                 
                     
                     
                 
                     
                         XKPCVKLTPLCVZ 
                 
                     
                     
                 
                     
                       XS-KPCVKLTPLCVZ 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
               
            
           
         
       
       in which-X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   ETSIKPCVKLTPLCVAMK 
                     
                 
                     
                     
                 
                     
                   DQSLKPCVKLTPLCVSLK 
                 
                     
                     
                 
                     
                       KPCVKLTPLCV 
                 
                     
                     
                 
                     
                     SLKPCVKLTPLCV 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         21 . A peptide according to  claim 20  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   ETSIKPCVKLTPLCVAMK 
                     
                 
                     
                     
                 
                     
                   DQSLKPCVKLTPLCVSLK 
                 
                     
                     
                 
                     
                   DQSLKPCVKLTPLCVTLN 
                 
                     
                     
                 
                     
                        PCVKLTPLC 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         22 . A peptide characterized as having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   X---N-S-IT--C-Z 
                     
                 
                     
                      XN-S-ITZ 
                 
                     
                     
                 
             
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   NHCNTSVITESCD 
                     
                 
                     
                      NTSVIT 
                 
                     
                     
                 
                     
                   TSCNTSVITQACP 
                 
                     
                      NTSAIT 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         23 . A peptide according to  claim 22  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   NHCNTSVITESCD 
                     
                 
                     
                      NTSVIT 
                 
                     
                     
                 
                     
                   TSCNTSVITQACP 
                 
                     
                      NTSVIT 
                 
                     
                     
                 
                     
                   INCNTSVITQACP 
                 
                     
                      NTSVIT 
                 
                     
                     
                 
                     
                   INCNTSAITQACP 
                 
                     
                      NTSAIT 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         24 . A peptide according to claim according to  claim 12  characterized as having the following formula:  
       XYC-P-G-A-L-C-N-TZ  
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of either of the following peptide sequences:  
       
         
           
                 
                 
               
                     
                 
                   11 YCAPPGYALLRC-NDT 
                     
                 
                     
                 
                   YCAPAGFAILKCNNKT 
                 
                     
                 
             
                
                
                
                
                
               
            
           
         
       
     
     
         25 . A peptide according to  claim 24  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   YCAPPGYALLRC-NDT 
                     
                 
                     
                     
                 
                     
                   YCAPAGFAILKCNNKT 
                 
                     
                     
                 
                     
                   YCAPAGFAILKCNDKK 
                 
                     
                     
                 
                     
                   YCAPAGFAILKCRDKK 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         26 . A peptide according to  claim 12  characterized as having the following formula:  
       X------A-C------W--Z  
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of either of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   NKRPRQAWCWFKG-KWKD 
                     
                 
                     
                     
                 
                     
                   N--MRQAHCNISRAKWNA 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
     
     
         27 . A peptide according to  claim 26  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   NKRPRQAWCWFKG-KWKT 
                     
                 
                     
                     
                 
                     
                   N--MRQAHCNISRAKWNA 
                 
                     
                     
                 
                     
                   D--IRRAYCTINETEWDK 
                 
                     
                     
                 
                     
                   I--IGQAHCNISRAQWSK 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         28 . A peptide according to  claim 12  characterized as having either of the following formulas:  
       
         
           
                 
                 
               
                     
                 
                   7 X-G-DPE------NC-GEF-YCN-----NZ 
                     
                 
                     
                 
                               XNC-GEF-YCNZ 
                 
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   KGSDPEVAYMWTNCRGEFLYCNMTWFLN 
                     
                 
                     
                     
                 
                     
                               NCRGEFLYCN 
                 
                     
                     
                 
                     
                   -GGDPEIVTHSFNCGGEFFYCNSTQLFN 
                 
                     
                     
                 
                     
                               NCGGEFFYCN 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         29 . A peptide according to  claim 28  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   KGSDPEVAYMWTNCRGEFLYCNMTWFLN 
                     
                 
                     
                     
                 
                     
                               NCRGEFLYCN 
                 
                     
                     
                 
                     
                   -GGDPEIVTHSFNCGGEFFYCNSTQLFN 
                 
                     
                     
                 
                     
                               NCGGEFFYCN 
                 
                     
                     
                 
                     
                   -GGDPEITTHSFNCRGEFFYCNTSKLFN 
                 
                     
                     
                 
                     
                               NCRGEFFYCN 
                 
                     
                     
                 
                     
                   -GGDPEITTHSFNCGGEFFYCNTSGLFN 
                 
                     
                     
                 
                     
                               NCGGEFFYCN 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         30 . A peptide according to  claim 12  characterized as having either of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   X-----C-IKQ-I------G---YZ 
                     
                 
                     
                     
                 
                     
                        XC-IKQ-IZ 
                 
                     
                     
                 
             
                
                
                
                
                
               
            
           
         
       
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of one of the following peptide sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   RNYAPCHIKQIINTWHKVGRNVY 
                     
                 
                     
                     
                 
                     
                        CHIKQII 
                 
                     
                     
                 
                     
                   TITLPCRIKQFINMWQEVGKAMY 
                 
                     
                     
                 
                     
                        CRIKQFI 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         31 . A peptide according to  claim 30  characterized as having one of the following formulas:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   RHYAPCHIKQIINTWHKVGRNVY 
                     
                 
                     
                     
                 
                     
                        CHIKQII 
                 
                     
                     
                 
                     
                   TITLPCRIKQFINMWQEVGKAMY 
                 
                     
                     
                 
                     
                        CRIKQFI 
                 
                     
                     
                 
                     
                   SITLPCRIKQIINMWQKTCKAMY 
                 
                     
                     
                 
                     
                        CRIKQII 
                 
                     
                     
                 
                     
                   NITLQCRIKQIIKMVAGR-KAIY 
                 
                     
                     
                 
                     
                        CRIKQII 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         32 . The antigenic peptide gag1 characterized as having the following formula:  
       XNCKLVLKGLGMNPTLEEMLTAZ  
       in which X and Z are OH or NH 2  or, to the extent that the immunological properties of the natural peptides lacking these groups shall not be essentially modified, the groups having from one to five amino acid residues, and each of the hyphens corresponding to an aminoacyl residue chosen from those which permit the conservation for the peptide characterized above of the immunological properties of the following peptide sequence:  
       XNCKLVLKGLGMNPTLEEMLTA  
     
     
         33 . An antigenic composition containing at least one gag1 peptide according to  claim 32  or at least an oligomer of this peptide, characterized as having the capacity to be recognized by human biological fluids such as serum containing anti-HIV-2 antibodies and under appropriate conditions anti-HIV-1 antibodies.  
     
     
         34 . An antigenic composition containing at least one peptide according to claims  13 ,  14  or  15 , or at least an oligomer of the peptide, characterized in that the peptide specifically recognizes the presence of anti-HIV-2 antibodies.  
     
     
         35 . An immunogenic composition containing at least one peptide according to any one of the claims  16 - 31  or at least an oligomer of the peptide or the peptide conjugated with a carrier molecule, in association with an acceptable pharmaceutical vehicle for the production of vaccines, the composition characterized in that it induces antibody production against the peptide in sufficient quantities to form an effective immunocomplex with the entire HIV-2 retrovirus and its corresponding proteins.  
     
     
         36 . An immunogenic composition according to  claim 35  further comprising peptides having formulas corresponding to the envelope glycloprotein sequences of HIV-1 and HIV-2 which have an amino acid homology greater than 50%.  
     
     
         37 . An immunogenic composition according to either of claims  35  or  36  having at least one peptide or at least an oligomer of the peptide or the peptide conjugated with a carrier molecule, the composition corresponding to a peptide chosen from the group consisting of Env4, Env5, Env6 and Env10.  
     
     
         38 . A procedure for the in vitro diagnosis-of HIV-2 infections in a biological fluid, comprising: 
 contacting the biological fluid with at least one peptide according to claims  12 ,  13 , 14 , 15  or  32 , or a conjugate of the peptide with a carrier molecule;    detecting the eventual presence in the biological fluid of an antigen-antibody complex by physical or chemical methods.    
     
     
         39 . The diagnostic procedure of  claim 38 , wherein the detection step is performed by a test selected by the group consisting of enzyme-linked immuno absorbant assay (ELISA), immunofluoresence assay (IFA), radioimmunoassay (RIA), and radioimmunoprecipitation assay (RIPIA).  
     
     
         40 . A kit for the in vitro diagnosis of an HIV-2 infection in a biological fluid comprising: 
 a peptide composition containing a peptide according to claims  12 ,  13 ,  14 ,  15  or  32 , or a mixture of such peptides, or a conjugate of such peptides with a carrier molecule;    an appropriate reaction environment for the production of an antigen-antibody complex;    one or more reagents adapted for the detection of the formation of antigen-antibody complexes; and    a biological fluid as a reference sample having no antibodies recognized by said peptide composition.    
     
     
         41 . An protein selected from the group described in Example 4 consisting of p 16, p 26, p 12, polymerase, Q protein, R protein, X protein, Y protein, env protein, F protein, TAT, ART, U5 and U3.  
     
     
         42 . A kit for diagnosing an HIV-2 infection by the method of  claim 6  and comprising as the polypeptide expression product a protein of  claim 41 .  
     
     
         43 . A vaccinating agent comprising at least one protein of  claim 41  in association with appropriate carriers.

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