HSV-2-DELTA-gD VACCINES AND METHODS FOR THEIR PRODUCTION AND USE

Abstract

Recombinant herpes simplex virus 2 (HSV-2) vaccine vectors, compositions and vaccines comprising such, and methods of use thereof are each provided.

1 . A process for producing a vaccine vector directed against a heterologous antigen, the process comprising:
 a) providing an HSV-2 genome comprising:
 (i) complete or partial deletion in a gene encoding HSV-2 glycoprotein D, and 
 (ii) a nucleic acid comprising a promoter-FP construct, wherein FP is a nucleic acid encoding a fluorescent protein; 
   b) co-transfecting a host cell with (i) the HSV-2 genome of a) and (ii) a linear DNA fragment encoding the heterologous antigen under conditions whereby allelic recombination occurs between the HSV-2 genome and the DNA fragment;   c) screening plaques resulting from b) to identify plaques not showing fluorescence under excitation light which elicits fluorescent protein fluorescence; and   d) recovering from those plaques not showing fluorescence in c) a recombinant HSV-2 virus or virion so as to obtain a vaccine vector directed against the heterologous antigen.   
     
     
         2 . The process of  claim 1 , wherein the host cell is a HSV-1 glycoprotein D complementing cell. 
     
     
         3 . The process of  claim 1 , wherein the promoter of the promoter-FP construct is a heterologous promoter. 
     
     
         4 . (canceled) 
     
     
         5 . The process of  claim 1 , wherein the fluorescent protein is Red Fluorescent Protein (RFP). 
     
     
         6 . The process of  claim 1 , wherein the host cell is co-transfected with
 the HSV-2 genome of a) and   a linear DNA fragment encoding, in order, HSV-2 gD signal sequence, the heterologous antigen, HSV-2 gD transmembrane domain, HSV-2 gD cytosolic domain, but not encoding a HSV-2 gD extracellular domain, or a linear DNA fragment encoding, in order, HSV-2 gD signal sequence, the heterologous antigen, and cytosolic domain of HSV-2 gD.   
     
     
         7 . The process of  claim 1 , wherein the host cell is co-transfected with the HSV-2 genome of a) and a linear DNA fragment encoding, in order, a promoter, the heterologous antigen, and optionally a poly-A signal. 
     
     
         8 . The process of  claim 1 , wherein the heterologous antigen is an influenza antigen, an HIV antigen, or a combination thereof. 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . The process of  claim 1 , wherein the heterologous antigen is under control of an upstream CMV promoter and has a downstream SV40 poly-A signal. 
     
     
         14 . The process of  claim 1 , wherein the promoter is a promoter of Elongation Factor 1a gene (P EF1α ) and wherein P EF1α  and FP are fused together (P EF1α -FP). 
     
     
         15 . A vaccine vector, or a recombinant herpes simplex virus-2 (HSV-2) comprising a genome encoding a heterologous antigen, made by the process of  claim 1 . 
     
     
         16 . A recombinant herpes simplex virus-2 (HSV-2) comprising:
 a complete or partial deletion of an HSV-2 glycoprotein D-encoding gene in the genome of the HSV-2; and   a linear DNA fragment encoding a promoter, a heterologous antigen signal sequence, and a heterologous antigen or a linear DNA fragment encoding a promoter, and a heterologous antigen.   
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . The recombinant HSV-2 of  claim 16 , wherein the heterologous antigen is an influenza antigen, an HIV antigen, or a combination thereof. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . A cell comprising therein the recombinant HSV-2 of  claim 16 , wherein the cell is not present in a human being. 
     
     
         26 . A vaccine composition comprising the recombinant HSV-2 of  claim 16 . 
     
     
         27 . A pharmaceutical composition comprising the recombinant HSV-2 of  claim 16 , and a pharmaceutically acceptable carrier. 
     
     
         28 . A method of eliciting and/or enhancing an immune response in a subject, the method comprising administering to the subject an amount of the recombinant HSV-2 of  claim 16 , in an amount effective to elicit and/or enhance the immune response in the subject. 
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . A method of treating or reducing the likelihood of an influenza infection in a subject, the method comprising administering to the subject an amount of the recombinant HSV-2 of  claim 16 , in an amount effective to treat or reduce the likelihood of an influenza infection in a subject, wherein the heterologous antigen is an influenza antigen. 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . A method of treating or reducing the likelihood of an HIV infection in a subject, the method comprising administering to the subject an amount of the recombinant HSV-2 of  claim 16 , in an amount effective to treat or reduce the likelihood of an HIV infection in a subject, wherein the heterologous antigen is an HIV antigen. 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . A method of vaccinating a subject for influenza infection, the method comprising administering to the subject an amount of the recombinant HSV-2 of  claim 16 , in an amount effective to vaccinate a subject for influenza infection, wherein the heterologous antigen is an influenza antigen. 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . A method of vaccinating a subject for HIV infection, the method comprising administering to the subject an amount of the recombinant HSV-2 of  claim 16 , in an amount effective to vaccinate a subject for HIV infection, wherein the heterologous antigen is an HIV antigen. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . A method of eliciting and/or enhancing an immune response in a subject, the method comprising administering to the subject an amount of a recombinant herpes simplex virus-2 (HSV-2) made by the process of  claim 1  and comprising (i) a complete deletion of an HSV-2 glycoprotein D-encoding gene in the genome thereof and (ii) encoding a promoter, an influenza hemagglutinin (HA) antigen signal sequence, and an HA antigen in an amount effective to elicit and/or enhance an immune response in a subject. 
     
     
         44 . A method of treating or reducing the likelihood of an influenza infection in a subject, the method comprising administering to the subject an amount of a recombinant herpes simplex virus-2 (HSV-2) made by the process of  claim 1  and comprising a complete or partial deletion of an HSV-2 glycoprotein D-encoding gene in the genome thereof and encoding a promoter, a influenza hemagglutinin (HA) antigen signal sequence, and an HA antigen, in an amount effective to treat or reduce the likelihood of an influenza infection in a subject. 
     
     
         45 . A method of vaccinating a subject for influenza infection, the method comprising administering to the subject an amount of a recombinant herpes simplex virus-2 (HSV-2) made by the process of  claim 1  and comprising a complete deletion of an HSV-2 glycoprotein D-encoding gene in the genome thereof; and encoding a promoter, an influenza hemagglutinin (HA) antigen signal sequence, and an HA antigen, in an amount effective to vaccinate a subject for influenza infection. 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . A method of quantitating a rate or amount of antibody-dependent cell-mediated killing (ADCK) in a population of cells, the method comprising:
 infecting a plurality of cells of the population of cells with a fluorescent protein-expressing recombinant HSV-2 that comprises a genome deleted for the gene encoding HSV-2 gD, under conditions permitting expression of the fluorescent protein in the cells,   contacting the plurality of infected cells with an antibody-containing solution and a population of immune cells, and   quantitating at one or more time points the amount of the plurality of infected cells exhibiting fluorescent protein fluorescence and, optionally, one or more markers, so as to quantitate over time the amount of live infected cells, so as to thereby quantitating the rate or amount of ADCK in the population of cells.   
     
     
         50 . (canceled) 
     
     
         51 . The method of  claim 49 , wherein the population of immune cells comprises macrophages. 
     
     
         52 . The method of  claim 49 , wherein the antibody-containing solution comprises serum. 
     
     
         53 . (canceled) 
     
     
         54 . (canceled) 
     
     
         55 . The method of  claim 49 , wherein the one or more markers comprises a cell membrane marker, a live/dead marker, or a combination thereof. 
     
     
         56 . The method of  claim 49 , further comprising quantitating at one or more time points the amount of cells exhibiting fluorescent protein fluorescence and, optionally, one or more markers, in a control population of infected cells otherwise identical but not contacted with an antibody-containing solution and comparing the amount or rate quantitated to the amount or rate quantitated for the population of cells contacted with the antibody-containing solution. 
     
     
         57 . (canceled) 
     
     
         58 . The method of  claim 49 , wherein the recombinant HSV-2 is made by a process comprising:
 a) providing an HSV-2 genome comprising:   (i) full or partial deletion in a gene encoding HSV-2 glycoprotein D, and   (ii) a nucleic acid comprising a promoter-FP construct, wherein FP is a nucleic acid encoding a fluorescent protein;   b) co-transfecting a host cell with the HSV-2 genome of a) and a linear DNA fragment encoding the heterologous antigen under conditions whereby allelic recombination occurs between the HSV-2 genome and the DNA fragment;   c) screening plaques resulting from b) to identify plaques not showing fluorescence under excitation light which elicits fluorescent protein fluorescence; and   d) recovering from those plaques not showing fluorescence in c) a recombinant HSV-2 viruses or virions so as to obtain a vaccine vector directed against the heterologous antigen.