Detection of CD8+ T cell responses to
View Patent ↗A method of detecting an anti-mycobacterial CD8 T cell response comprising contacting a population of CD8 T cells of an individual with one or more peptides selected from the peptides represented by SEQ ID NO: 3, 4, 7, 8, 9, 10, 11 or 12, and, optionally, one or two further peptides represented by SEQ ID NO: 1 and/or 2, wherein one or more of said peptides may be substituted by an analogue which binds a T cell receptor which recognises the corresponding substituted peptide, and determining whether CD8 T cells of the CD8 T cell population recognize the peptide(s). The invention also provides a method of vaccinating against infection by a mycobacterium , wherein the vaccination leads to a CD8 T cell response, comprising administering (i) a CD8 T cell epitope of a mycobacterium protein, (ii) an analogue of the epitope which is capable of inhibiting the binding of the epitope to a T cell receptor, (iii) a precursor or (i) or (ii) which is capable of being processed to provide (i) or (ii), or (iv) a polynucleotide which is capable of being expressed to provide (i), (ii) or (iii).
1. A method of detecting an anti-mycobacterial CD8 T cell response comprising contacting a population of CD8 T cells of a human individual with one or more peptides selected from the group consisting of SEQ ID NOS:3, 4, 7, 8, 9, 10, 11 and 12, and, optionally, one or two further peptides selected from the group consisting of SEQ ID NOS:1 and 2, wherein one or more peptides may be substituted by an analogue which binds a T cell receptor that recognizes the peptide, and determining whether CD8 T cells of the CD8 T cell population recognize the peptide(s).
2. A method according to claim 1 wherein a peptide panel is employed, wherein said panel includes peptides consisting of SEQ ID NOS:1, 2, 3, 4, 8, 9 and 10, wherein one or more peptides may be substituted by said analogue.
3. A method according to claim 1 wherein any analogue which is used is (i) at least 70% homologous, preferably at least 80% homologous, more preferably at least 90% homologous, to the entire peptide, and/or (ii) has one or more deletions at the N-terminus and/or C-terminus in comparison to the peptide, and/or (iii) has one or more conservative substitutions compared to the peptide.
4. A method according to claim 1 in which the recognition of the peptide(s) by the CD8 T cells is determined by measuring secretion of a cytokine from the CD8 T cells.
5. A method according to claim 4 in which IFN-γ secretion from the T cells is measured.
6. A method according to claim 5 in which IFN-γ secretion from the CD8 T cells is determined by allowing secreted IFN-γ to bind an immobilized antibody specific to the cytokine and then determining the presence of antibody/cytokine complex.
7. A method according to claim 1 in which the CD8 T cells are freshly isolated ex vivo cells from peripheral blood.
8. A method according to claim 1 in which CD8 T cells are pre-cultured in vitro with the peptide(s).
9. A method according to claim 1 in which the mycobacterium is M. tuberculosis.
10. A method according to claim 1 wherein the population of CD8 T cells is from an individual to whom an anti-mycobacterial vaccine has been administered.
11. A method according to claim 1 which is carried out in vitro.
12. A kit comprising one or more peptides selected from the group consisting of SEQ ID NOS:3, 8, 9, 10, 11 and 12, and, optionally, one or two further peptides selected from the group consisting of SEQ ID NOS:1 and 2, wherein one or more peptides may be substituted by an analogue which binds a T cell receptor which recognizes the peptide, and optionally a means to detect recognition of the peptide(s) by CD8 T cells.
13. A kit comprising a peptide panel, wherein said panel includes peptides consisting of SEQ ID NOS:1, 2, 3, 4, 8, 9 and 10, wherein one or more peptides may be substituted by an analogue which binds a T cell receptor which recognizes the peptide.
14. A kit according to claim 12 which includes an antibody to IFN-γ.
15. A kit according to claim 14 wherein said antibody is immobilized on a solid support and which optionally also includes a means to detect any antibody/IFN-γ complex.
16. A kit according to claim 12 which includes the means to detect recognition of the peptide(s) by CD8 T cells.
17. A kit according to claim 14 which includes the means to detect recognition of the peptide(s) by CD8 T cells.
18. A kit according to claim 15 which includes the means to detect any antibody/IFN-γ complex.