DNA-vaccines based on constructs derived from the genomes of human and animal pathogens

Abstract

Methods of eliciting an immune response in a subject by administering one or more large genomic DNA fragments are provided. Also provided are methods of identifying sequences encoding antigenic polypeptides. Also provided are vaccine compositions comprising one or more large genomic DNA fragments.

1 . A vaccine, comprising a construct that comprises one or more pathogenic DNA genomic fragments, wherein (a) the collective size of the fragment(s) is about 5-50 Kb and (b) at least one fragment comprises at least one antigen coding sequence.  
     
     
         2 . The vaccine of  claim 1 , wherein the pathogenic DNA fragments are from a virus pathogen.  
     
     
         3 . The vaccine of  claim 2 , wherein the virus is a herpes virus.  
     
     
         4 . The vaccine of  claim 3 , wherein the herpes virus is selected from the group consisting of a Herpes Simplex Virus (HSV), Varicella Zoster Virus (VZV), Epstein Barr Virus (EBV) and a Cytomegalovirus (CMV).  
     
     
         5 . The vaccine of  claim 4 , wherein the HSV virus is herpes simplex virus-2 (HSV-2).  
     
     
         6 . The vaccine of  claim 4 , wherein the virus is a Herpes Simplex Virus and the HSV genomic DNA fragment(s) express HSV antigens consisting essentially of those encoded by the HSV immediate early genes ICP 27, ICP 0, ICP 4 and ICP 22.  
     
     
         7 . The vaccine of  claim 6 , wherein (i) the HSV genomic DNA fragment(s) are from HSV-2 and (ii) correspond to the sequence extending from the 7th to 10th EcoR1 sites shown in  FIG. 1 .  
     
     
         8 . The vaccine of  claim 1 , wherein the vaccine comprises multiple different constructs.  
     
     
         9 . The vaccine of  claim 8 , wherein the constructs are coated onto core carriers.  
     
     
         10 . The vaccine of  claim 9 , wherein the core carriers have an average diameter of about 0.5 to about 5 μm and a density sufficient to allow delivery into a subject.  
     
     
         11 . The vaccine of  claim 10 , wherein the core carrier is made of metal.  
     
     
         12 . The vaccine of  claim 11 , wherein the metal is gold.  
     
     
         13 . The vaccine of  claim 1 , wherein the construct is either (A) (i) a plasmid and (ii) the collective size of the fragment(s) is about 5-25 Kb, or (B) (i) a cosmid and (ii) the collective size of the fragment(s) is about 25-50 Kb.  
     
     
         14 . The vaccine of  claim 1 , wherein the antigen coding sequence(s) are not expressed by heterologous promoters.  
     
     
         15 . A method for eliciting an immune response in a vertebrate subject, comprising administering to a vertebrate subject the vaccine of  claim 1 , whereby expression of the antigen coding sequence(s) of the construct elicits an immune response against the expressed antigen.  
     
     
         16 . The method of  claim 15 , wherein (i) the constructs of the vaccine are coated onto core carriers and (ii) the vaccine is administered to the vertebrate subject using a particle-mediated transdermal delivery technique.  
     
     
         17 . The method of  claim 15 , further comprising repeating the administration to provide a prime and a boost.  
     
     
         18 . The method of  claim 15 , wherein the vaccine is in a formulation that comprises an adjuvant  
     
     
         19 . The method of  claim 15  further comprising administering an adjuvant.  
     
     
         20 . A method of eliciting an immune response in an individual, comprising administering constructs to an individual, wherein the constructs comprise pathogenic genomic fragments that (i) comprise an array of antigen-encoding sequence, but which (ii) do not comprise sequences that encode immunodominant antigens or genes whose products inhibit immune responses.