Rapid, efficient purification of HSV-specific T-lymphocytes and HSV antigens identified via same
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 antigen or polynucleotides encoding antigens of the invention provide effectively targeted vaccines for prevention and treatment of HSV infection.
1. A pharmaceutical composition comprising a herpes simplex virus type 2 (HSV-2) US6 polypeptide, wherein the HSV-2 US6 polypeptide consists of amino acids 365-373 or 342-393 and up to 30% of the full length of US6 (SEQ ID NO: 11).
2. The pharmaceutical composition of claim 1 , wherein the polypeptide is a fusion protein comprising the HSV-2 polypeptide fused to a heterologous polypeptide.
3. The pharmaceutical composition of claim 2 , wherein the fusion protein is soluble.
4. The pharmaceutical composition of claim 2 , 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.
5. The pharmaceutical composition of claim 1 , further comprising an adjuvant.
6. The pharmaceutical composition of claim 1 , wherein the polypeptide comprises up to 20% of the full length of US6 (SEQ ID NO: 11).
7. The pharmaceutical composition of claim 1 , wherein the polypeptide comprises up to 10% of the full length of US6 (SEQ ID NO: 11).
8. A pharmaceutical composition comprising a herpes simplex virus type 2 (HSV-2) polypeptide or a polynucleotide encoding said polypeptide, wherein the polypeptide consists of amino acids 365-373 or 342-393 of US6 (SEQ ID NO: 11).
9. The pharmaceutical composition of claim 8 , wherein the polynucleotide comprises a recombinant virus genetically modified to express amino acids 365-373 or 342-393 of US6 (SEQ ID NO: 11).
10. A pharmaceutical composition comprising a polynucleotide that encodes a herpes simplex virus type 2 (HSV-2) polypeptide consisting of amino acids 365-373 or 342-393 and up to 30% of the full length of US6 (SEQ ID NO: 11).
11. The pharmaceutical composition of claim 10 , wherein the polypeptide comprises up to 20% of the full length of US6 (SEQ ID NO: 11).
12. The pharmaceutical composition of claim 10 , further comprising an adjuvant.
13. The pharmaceutical composition of claim 10 , wherein the polypeptide is a fusion protein comprising the HSV-2 polypeptide fused to a heterologous polypeptide.
14. The pharmaceutical composition of claim 10 , wherein the polynucleotide comprises a recombinant virus genetically modified to express the HSV-2 polypeptide.
15. The pharmaceutical composition of claim 14 , wherein the recombinant virus is a vaccinia virus, canary pox virus or adenovirus.
16. The pharmaceutical composition of claim 14 , further comprising an adjuvant.
17. A method of inducing an immune response to an HSV infection in a subject comprising administering the composition of claim 1 to the subject.
18. A method of inducing an immune response to an HSV infection in a subject comprising administering the composition of claim 10 to the subject.
19. A method of treating an HSV infection in a subject comprising administering the composition of claim 1 to the subject.
20. A method of treating an HSV infection in a subject comprising administering the composition of claim 10 to the subject.