Vectors and methods for immunization or therapeutic protocols

Abstract

The present invention shows that DNA vaccine vectors can be improved by removal of CpG-N motifs and optional addition of CpG-S motifs. In addition, for high and long-lasting levels of expression, the optimized vector should include a promoter/enhancer that is hot down-regulated by the cytokines induced by the immunostimulatory CpG motifs. Vectors and methods of use for immunostimulation are provided herein. The invention also provides improved gene therapy vectors by determining the CpG-N and CpG-S motifs present in the construct, removing stimulatory CpG (CpG-S) motifs and/or inserting neutralizing CpG (CpG-N) motifs, thereby producing a nucleic acid construct providing enhanced expression of the therapeutic polypeptide. Methods of use for such vectors are also included herein.

1 - 58 . (Canceled)  
     
     
         59 . A method for enhancing the expression of a polynucleotide in vivo wherein the polynucleotide is contained in a nucleic acid construct, the method comprising, determining the CpG-N and CpG-S motifs present in the construct, removing stimulatory CpG (CpG-S) motifs and/or inserting neutralizing CpG (CpG-N) motifs, thereby producing a nucleic acid construct providing enhanced expression of the polynucleotide.  
     
     
         60 - 82 . (Canceled)  
     
     
         83 . The method of  claim 59 , wherein the polynucleotide is an antisense polynucleotide or a ribozyme.  
     
     
         84 . A method for enhancing the expression of a polynucleotide in vivo, the method comprising administering to a subject a nucleic acid construct, wherein the construct is produced by determining the CpG-N and CpG-S motifs present in the construct and removing stimulatory CpG (CpG-S) motifs and/or inserting neutralizing CpG (CpG-N) motifs, thereby enhancing expression of a polynucleotide expressed by the nucleic acid construct in the subject.  
     
     
         85 - 107 . (Canceled)  
     
     
         108 . The method of  claim 84 , wherein the polynucleotide is an antisense polynucleotide or a ribozyme.