T-cell reactivity platform

1. A method for assaying antigen-specific T-cell activation, comprising the steps of:

a. providing a phagocytable particle, having a candidate antigen polypeptide associated by covalent linking, wherein the particle with the associated polypeptide has been subjected to a denaturing wash resulting in an endotoxin level low enough to not interfere with steps b-f;

b. providing a viable antigen-presenting cell;

c. contacting the washed particle with the antigen-presenting cell in vitro under conditions allowing phagocytosis of the particle by the antigen-presenting cell;

d. providing a T-cell sample to be assayed, the T-cell sample comprising viable T-cells;

e. contacting the T-cell sample with the antigen-presenting cell contacted with the particle in vitro under conditions allowing specific activation of T-cells in response to an antigen presented by an antigen-presenting cell; and

f. determining the degree of T-cell activation in the T-cell sample.

2. A method for assaying antigen-specific T-cell activation, comprising the steps of:

a. providing a phagocytable particle;

b. associating by covalent linking a candidate antigen polypeptide to the particle;

c. subjecting the particle with the associated polypeptide to a denaturing wash resulting in an endotoxin level low enough to not interfere with steps d-h;

d. providing a viable antigen-presenting cell;

e. contacting the washed particle with the antigen-presenting cell in vitro under conditions allowing phagocytosis of the particle by the antigen-presenting cell;

f. providing a T-cell sample to be assayed, the T-cell sample comprising viable T-cells;

g. contacting the T-cell sample with the antigen-presenting cell contacted with the particle in vitro under conditions allowing specific activation of T-cells in response to an antigen presented by an antigen-presenting cell; and

h. determining the degree of T-cell activation in the T-cell sample.

3. The method according to claim 1 , further comprising the step of comparing the degree of T-cell activation to a relevant reference, whereby a higher degree of T-cell activation in the sample compared to the reference indicates that the candidate antigen results in antigen-specific T-cell activation in the sample.

4. The method according to claim 1 , wherein determining the degree of T-cell activation in the T-cell sample comprises determining the fraction of activated T-cells in the sample.

5. The method according to claim 1 , wherein several candidate antigens are assayed against the same T-cell sample.

6. The method according to claim 5 , wherein at least 10 candidate antigens are assayed against the same T-cell sample.

7. The method according to claim 1 , wherein the antigen-presenting cell and the T-cell sample are derived from a single individual.

8. The method according to claim 7 , wherein the antigen-presenting cell and the T-cell sample are derived from the same blood sample from the individual.

9. The method according to claim 7 , wherein the antigen-presenting cell and the T cell sample are derived from a peripheral blood mononuclear cell (PBMC) sample from the individual.

10. The method according to claim 1 , wherein the T-cell sample is derived from a tumour.

11. The method according to claim 1 , wherein the T-cell sample is derived from ascites.

12. The method according to claim 1 , wherein the T-cell sample comprises CD4 + T-cells, CD8 + T-cells, or both.

13. The method according to claim 1 , wherein the washed particle, the antigen presenting cell and the T-cell sample are contacted concurrently.

14. The method according to claim 1 , wherein determining the degree of T-cell activation in the T-cell sample comprises determining the fraction of T-cells contacted with the antigen-presenting cell responding by secretion of IFN-γ, IL-17, IL-22, or combinations thereof.

15. The method according to claim 1 , wherein determining the degree of T-cell activation in the T-cell sample comprises determining the fraction of T-cells contacted with the antigen-presenting cell responding by secretion of IL-17, IL-22, or both.

16. The method according to claim 1 , wherein the candidate antigen polypeptide comprises at least 50 amino acids.

17. The method according to claim 1 , wherein the T-cell sample is from a human and the candidate antigen polypeptide sequence is derived from a human.

18. The method according to claim 1 , wherein the candidate antigen polypeptide is derived from a polypeptide:

a. known to be highly expressed in a tissue or cell affected in an autoimmune disease;

b. known to be associated with a neoplastic disease;

c. known to be associated with an autoimmune disease;

d. known to be associated with an infectious disease; or

e. known to be associated with allergy or similar hypersensitivity.

19. The method according to claim 1 , comprising the steps of:

a. providing a phagocytable particle, having a candidate peptide auto-antigen associated by covalently linking, wherein the particle with the associated polypeptide has been subjected to a denaturing wash resulting in an endotoxin level low enough to not interfere with steps b-e;

b. providing a peripheral blood mononuclear cell (PBMC) sample from a test subject, the PBMC sample comprising viable T-cells and viable antigen-presenting cells;

c. contacting the PBMC sample with the particle in vitro under conditions allowing phagocytosis of the particle by an antigen-presenting cell and allowing specific activation of T-cells in response to an antigen presented by an antigen-presenting cell;

d. quantitating the fraction of activated T-cells in the contacted PBMC sample; and

e. comparing the quantitated fraction to a comparable quantitated fraction from a healthy subject, wherein a higher fraction of activated T-cells in the test subject as compared to the healthy subject is indicative of autoreactivity against the candidate auto-antigen in the test subject.