Rapid, efficient purification of hsv-specific t-lymphocytes and hsv antigens identified via same

Abstract

Described is a method of identifying an immunologically active antigen of a virus that attacks skin, as well as a method of enriching a population of lymphocytes for T lymphocytes that are specific to a virus that attacks skin. Also provided are HSV antigens and epitopes that are useful for the prevention and treatment of HSV infection that have been identified via the methods of the invention. T-cells having specificity for antigens of the invention have demonstrated cytotoxic activity against cells loaded with virally-encoded peptide epitopes, and in many cases, against cells infected with HSV. The identification of immunogenic antigens responsible for T-cell specificity provides improved anti-viral therapeutic and prophylactic strategies. Compositions containing antigens or polynucleotides encoding antigens of the invention provide effectively targeted vaccines for prevention and treatment of HSV infection.

What is claimed is: 
     
         1 . A pharmaceutical composition comprising a herpes simplex virus type 2 (HSV-2) US6 polypeptide or a polynucleotide encoding same, wherein the HSV-2 US6 polypeptide consists of amino acids 365-373 (SEQ ID NO: 5) and up to 90% of the full length of US6 (SEQ ID NO: 11). 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the HSV-2 US6 polypeptide consists of the C-terminal cytoplasmic tail of US6 and up to 90% of the full length of US6 (SEQ ID NO: 11). 
     
     
         3 . The pharmaceutical composition of  claim 2 , wherein the C-terminal cytoplasmic tail comprises amino acids 365-393 of SEQ ID NO: 11. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the polypeptide comprises amino acids 365-373 and contiguous adjacent sequence of SEQ ID NO: 11, wherein the contiguous adjacent sequence is up to 10% of the full length of US6 (SEQ ID NO: 11). 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the polypeptide is a fusion protein comprising the HSV polypeptide fused to a heterologous polypeptide. 
     
     
         6 . The pharmaceutical composition of  claim 2 , wherein the fusion protein is soluble. 
     
     
         7 . The pharmaceutical composition of  claim 5 , wherein the heterologous polypeptide is selected from the group consisting of UL7 (SEQ ID NO: 7), UL25 (SEQ ID NO: 8), UL26 (SEQ ID NO: 9), UL46 (SEQ ID NO: 10), or US8 (SEQ ID NO: 12) of HSV-2. 
     
     
         8 . The pharmaceutical composition of  claim 1 , further comprising an additional HSV polypeptide. 
     
     
         9 . The pharmaceutical composition of  claim 8 , wherein the additional HSV polypeptide is selected from the group consisting of UL7 (SEQ ID NO: 7), UL25 (SEQ ID NO: 8), UL26 (SEQ ID NO: 9), UL46 (SEQ ID NO: 10), or US8 (SEQ ID NO: 12) of HSV-2. 
     
     
         10 . The pharmaceutical composition of  claim 1 , further comprising an adjuvant. 
     
     
         11 . The pharmaceutical composition of  claim 1 , wherein the polynucleotide comprises a recombinant virus genetically modified to express the HSV-2 US6 polypeptide. 
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein the recombinant virus is a vaccinia virus, canary pox virus or adenovirus. 
     
     
         13 . A method of inducing an immune response to an HSV infection in a subject comprising administering the composition of  claim 1  to the subject. 
     
     
         14 . The method of  claim 12 , wherein the immune response comprises a CD8 response. 
     
     
         15 . The method of  claim 13 , wherein the CD8 response is detected by measuring IFNγ release, proliferation or cytotoxicity of CD8-positive T cells. 
     
     
         16 . A method of treating an HSV infection in a subject comprising administering the composition of  claim 1  to the subject.