Detection of individual T-cell reaction patterns against tumor-associated antigens (TAA) in tumor patients as a basis for the individual therapeutic vaccination of patients
The present invention relates to a method for identifying the preferential target antigens of antitumoural T-cells of a tumour patient, comprising: a) providing T-cells from the blood of at least one tumour patient, b) providing dendritic cell (DCs) and/or B-lymphocytes (BLCs) that are autologous for said tumour patient, wherein said DCs and BLCs were transfected beforehand with a selection of mRNAs encoding for T-cell-immunogenic tumour-associated antigens (TAA), and express these, c) contacting said T-cells with the DCs and/or BLCs, d) identifying of those T-cells that recognize antigens of the DCs and/or BLCs, and e) identifying of the preferential target antigens of antitumoural T-cells of the at least one tumour patient on the basis of the T-cells that recognize antigens of the DCs and/or BLCs. The method can furthermore comprise the expansion of the T-cells that recognize the antigens of the DCs and/or BLCs. The present invention furthermore relates to a method for producing an individualized tumour vaccine or individualized tumour therapeutic, as well as corresponding methods for treating a tumourous disease using the individualised tumour vaccine or individualised tumour therapeutic.
1. A method for identifying the preferential target antigens of anti-tumoural T-cells of at least one tumour patient that is suffering from a specific tumour comprising:
a) providing T cells from the blood of said at least one tumour patient, wherein the T cells are not stimulated by contact with a tumour cell line in vitro;
b) providing a panel of transfected dendritic cells (DCs) and/or a panel of transfected B-lymphocytes (BLCs) for said at least one tumour patient, wherein said panel of DCs and/or BLCs are autologous DCs or autologous BLCs transfected with mRNA encoding individual T-cell tumour antigens associated with said specific tumour, wherein multiple T cell tumour antigens are represented within the panel, and wherein said mRNA is provided independent from the extraction of tumour cells from said at least one tumour patient;
c) contacting the panel of transfected dendritic cells (DCs) and/or transfected B-lymphocytes (BLCs) for said at least one tumour patient with the respective autologous T cells; and
d) identifying the tumour-associated antigens (TAAs) that are recognized by the T cells on the panel of transfected DCs and/or BLCs for said at least one tumour patient.
2. The method according to claim 1 , further comprising the expansion of the T-cells that recognize antigens of the DCs and/or BLCs.
3. The method according to claim 1 , further comprising a control assay for the reactivity of the T-cells against the TAA.
4. The method according to claim 1 further comprising determining the HLA alleles by which the recognized TAAs are presented.
5. The method according to claim 1 wherein the T cells are isolated T cells from the blood of said at least one tumour patient.
6. The method according to claim 1 wherein the mRNA encoding the tumour antigens are mRNA encoding antigens of several categories.
7. The method according to claim 6 , wherein the antigens are melanocyte-specific proteins or cancer/germline-proteins (C/G-proteins).
8. The method according to claim 6 wherein the categories of tumour antigens are selected from the group consisting of differentiation antigens, cancer/germline antigens (C/G-antigens), mutated antigens and over-expressed antigens.
9. The method according to claim 8 wherein the mutated antigen is a fusion protein.
10. The method according to claim 1 wherein the specific tumour is malignant melanoma.
11. The method according to claim 6 , wherein the mRNA encoding the tumour antigens includes structurally normal, “shared” TAAs.
12. The method according to claim 11 , wherein said structurally normal (“shared”) TAAs are selected from the group consisting of BAGE-1; GAGE-1,2,8; GAGE-3,4,5,6,7; GnTV (intron); HER V-K-MEL; KK-LC-1; KM-HN-1; LAGE-1; MAGE-A1; MAGE-A2; MAGE-A3; MAGE-A4; MAGE-A6; MAGE-A9; MAGE-A10; MAGE-A12; MAGE-C2; mucin; NA-88; NY-ESO-1/LAGE-2; SAGE; Sp17; SSX-2, SSX-4; and TRP2-INT2(intron 2).
13. The method according to claim 1 wherein the specific tumour is a tumour selected from the group consisting of kidney, breast, pancreatic, stomach, testicular, prostate, colon and skin.