Identification of gene sequences and proteins involved in vaccinia virus dominant T cell epitopes
View Patent ↗The present invention relates to the identification of gene sequences and proteins involved in vaccinia virus dominant T cell epitopes. Two vaccinia virus CD8 + T cell epitopes restricted by the most common human MHC class I allele, HLA-A0201 have been identified. Both epitopes are highly conserved in vaccinia and variola viruses. The induction of the T cell responses following primary vaccination is demonstrated by the kinetics of epitope specific CD8 + T cells in 3 HLA-A0201 individuals. This information will be useful for the design and analyses of the immunogenicity of experimental vaccinia vaccines, and for basic studies of human T cell memory.
1. A method of identifying T cells in a sample that become activated in the presence of a vaccinia or variola virus that comprises a polypeptide having an amino acid sequence that is identical or substantially homologous to peptide 165 (SEQ ID NO: 2), comprising contacting the T cells with a polypeptide selected from the group consisting of:
a) peptide 165 (SEQ ID NO: 2),
b) an immunogenic mutant or fragment of SEQ ID NO: 2, wherein the immunogenic mutant or fragment maintains the function of peptide 165 as a CD8 T cell epitope of the vaccinia or variola virus, and
c) a combination thereof,
wherein activation of the T cells by the polypeptide indicates that the T cells become activated in the presence of the vaccinia or variola virus.
2. The method of claim 1 wherein whether the T cells present in the sample become activated is determined using an assay selected from the group consisting of: a cytokine assay, a flow cytometry assay and a limiting dilution assay.
3. The method of claim 2 wherein the cytokine assay is an ELISPOT assay and the flow cytometry assay is a tetramer staining assay.
4. The method of claim 1 wherein the sample is selected from the group consisting of: blood, lymph and tissue.
5. The method of claim 4 wherein the sample is a peripheral blood mononuclear cell sample.
6. A method of determining whether an individual has been infected with a vaccinia or variola virus that comprises a polypeptide having an amino acid sequence that is identical or substantially homologous to peptide 165 (SEQ ID NO: 2), comprising determining whether the individual's T cells become activated in the presence of a polypeptide selected from the group consisting of: peptide 165 (SEQ ID NO: 2), an immunogenic mutant or fragment of SEQ ID NO: 2, wherein the immunogenic mutant or fragment maintains the function of peptide 165 as a CD8 T cell epitope of the vaccinia or variola virus, and a combination thereof, wherein if the individual's T cells become activated in the presence of the peptide, then the individual has been infected with the vaccinia or variola virus.
7. The method of claim 6 wherein the individual's T cells are present in a sample, and the sample is selected from the group consisting of: blood, lymph and tissue.
8. The method of claim 7 wherein the sample is a peripheral blood mononuclear cell sample.
9. The method of claim 6 wherein whether the whether the individual's T cells become activated is determined using an assay selected from the group consisting of: a cytokine assay, a flow cytometry assay and a limiting dilution assay.
10. The method of claim 9 wherein the cytokine assay is an ELISPOT assay and the flow cytometry assay is a tetramer staining assay.
11. A method of monitoring the effectiveness of a vaccinia vaccine that comprises a polypeptide having an amino acid sequence that is identical or substantially homologous to peptide 165 (SEQ ID NO: 2) in an individual who has been administered the vaccinia vaccine, comprising determining whether the individual's T cells become activated in the presence of a polypeptide selected from the group consisting of: peptide 165 (SEQ ID NO: 2), an immunogenic mutant or fragment of SEQ ID NO: 2, wherein the immunogenic mutant or fragment maintains the function of peptide 165 as a CD8 T cell epitope of the vaccinia or variola virus, and a combination thereof, wherein if the individual's T cells become activated, then the vaccinia vaccine is effective in the individual.
12. The method of claim 11 wherein the individual's T cells are present in a sample, and the sample is selected from the group consisting of: blood, lymph and tissue.
13. The method of claim 12 wherein the sample is a peripheral blood mononuclear cell sample.
14. The method of claim 11 wherein whether the whether the individual's T cells become activated is determined using an assay selected from the group consisting of: a cytokine assay, a flow cytometry assay and a limiting dilution assay.
15. The method of claim 14 wherein the cytokine assay is an ELISPOT assay and the flow cytometry assay is a tetramer staining assay.
16. The method of claim 11 wherein the vaccinia vaccine is a cancer vaccine.