Process for the production of HCMV glycoproteins, antibodies thereto and HCMV vaccines, and recombinant vectors therefor

Abstract

HCMV glycoproteins B and H have been identified. The gB protein is encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary. The gH protein is encoded by DNA in the HindIII L fragment lying between 228 and 2456 bases from the L/D boundary. The genes have been incorporated in recombinant vaccinia vectors and expressed in host animals to raise HCMV-neutralising antibody, thereby indicating vaccine potential. The glycoproteins can also be used in a variety of different ways, as vaccines or in the production, purification or detection of HCMV antibody.

1 . A process which comprises expressing from a recombinant DNA vector in a suitable host organism a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary.  
     
     
         2 . A process according to  claim 1  in which said polypeptide is incorporated into a vaccine against HCMV in humans.  
     
     
         3 . A recombinant virus vector containing DNA encoding a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary, said vector being capable of infecting a human subject and expressing said polypeptide in immunogenic form.  
     
     
         4 . A vaccine against HCMV, incorporating a recombinant virus vector of  claim 3 .  
     
     
         5 . A DNA isolate which encodes a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary.  
     
     
         6 . A process which comprises synthesising a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary.  
     
     
         7 . A method of preparing HCMV monospecific antiserum, which comprises immunising a host animal with a polypeptide prepared by a process of  claim 1  or  claim 6  or with a recombinant virus vector of  claim 3 , and extracting from the host animal antiserum specific to said polypeptide.  
     
     
         8 . A method which comprises immunising a host animal with a polypeptide prepared by a process of  claim 1  or  claim 6  or with a recombinant virus vector of  claim 3 , and preparing HCMV-specific monoclonal antibody from cells of the animal thus immunised.  
     
     
         9 . A method of purifying HCMV-specific antibodies, which comprises contacting the antibodies with a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary, and separating bound antibody from said polypeptide.  
     
     
         10 . A method of detecting HCMV-specific antibody in a clinical sample, which comprises contacting the sample with a polypeptide incorporating one or more antigenic determinants capable of raising HCMV-neutralising antibodies in humans, said determinant or determinants corresponding to a portion of the protein encoded by DNA in the HindIII F fragment of the HCMV genome lying between 1378 and 4095 bases from the F/D boundary and/or a portion of the protein encoded by DNA in the HindIII L fragment of the HCMV genome lying between 228 and 2456 bases from the L/D boundary, and detecting antibody that binds to said polypeptide.  
     
     
         11 . A kit for carrying out the method of  claim 10 , said kit comprising said polypeptide in a form suitable for contacting with the clinical sample, and means for detecting HCMV-specific antibody that binds to said polypeptide.