Method for treating disseminated cancer

1. A method for treating a patient suffering from a disseminated cancer, the method comprising:

i) identifying in a patient one or more metinel lymph nodes, and, optionally, identifying all or part of metastases of the disseminated cancer;

ii) resecting the one or more nodes and, optionally, all or part of the metastases;

iii) isolating metastasis-reactive T-lymphocytes from said lymph nodes;

iv) in vitro expanding said metastasis-reactive T-lymphocytes; and

v) administering the thus obtained T-lymphocytes to the patient,

wherein the T-lymphocytes are CD4+helper or CD8+T-lymphocytes.

2. A method according to claim 1 , wherein the in vitro expansion step comprises:

(a) a first phase of stimulating tumour-reactive CD4+helper or CD8+T-lymphocytes with tumour-derived antigen together with at least one substance having agonistic activity towards the IL-2 receptor to promote survival of tumour-reactive CD4+helper or CD8+T-lymphocytes; and

(b) a second phase of activating and promoting growth of tumour-reactive CD4+helper or CD8+T-lymphocytes, wherein the second phase is initiated when the CD25 cell surface marker or IL-2R marker is down-regulated on T-lymphocytes.

3. A method according to claim 2 , wherein the down-regulation is defined as that 5% or less of the T-lymphocyte population expresses CD25.

4. A method according to claim 3 , wherein the first phase is initiated by adding the at least one substance having agonistic activity towards the IL-2 receptor.

5. A method according to claim 4 , wherein the substance having agonistic activity towards the IL-2 receptor is IL-2.

6. A method according to claim 2 , wherein the tumour-derived antigen is a denatured homogenate of a tumour.

7. A method according to claim 2 , wherein the second phase is initiated by the addition of tumour-derived antigen to the T-lymphocytes for activating tumour-reactive CD25-negative T-lymphocytes.

8. A method according to claim 7 , which further comprises addition of antigen presenting cells to the T-lymphocytes together with the tumour-derived antigen.

9. A method according to claim 8 , wherein the antigen presenting cells are irradiated peripheral blood leucocytes containing antigen-presenting B-cells and/or monocytes.

10. A method according to claim 2 , wherein the second phase comprises adding at least one substance capable of up-regulating IL-12R on the T-lymphocytes.

11. A method according to claim 2 , wherein the second phase comprises adding one or more substances capable of antagonizing development of Th2 type T-lymphocytes.

12. A method according to claim 11 , wherein the one or more substances capable of antagonizing development of Th2 type T-lymphocytes are one or more substances capable of neutralizing IL-4, IL-5, IL-10, or TGF-beta.

13. A method according to claim 12 , wherein the one or more substances capable of neutralizing IL-4, IL-5, IL-10, or TGF-beta are anti IL-4 antibody, anti IL-5 antibody or anti IL-10 antibody.

14. A method according to any of the claim 2 , 12 or 13 , wherein a further amount of the one or more substance capable of antagonizing development of Th2 type T-lymphocytes is added regularly throughout phase.

15. A method according to claim 2 , wherein the second phase comprises adding one or more substances promoting the development of Th1 type T-lymphocytes.

16. A method according to claim 15 , wherein the one or more substances promoting the development of Th1 type T-lymphocytes is substances having agonistic activity towards the IL-7, IL-12, IL-15 or IL-21 receptor.

17. A method according to claim 16 , wherein the one or more substances is selected from IL-7, IL-12, IL-15 and IL-21.

18. A method according to either of claim 1 or 2 for the preparation of Th1 type T-lymphocytes of the memory or effector type

19. A method according to claim 2 , which further comprises monitoring the expression of cell surface markers continuously during the first phase and second phase, and wherein the T-lymphocytes are harvested when CD25 on T-lymphocytes in the second phase is down-regulated.

20. A method according to claim 19 , wherein the T-lymphocytes are subjected to at least one additional round of the second phase, when CD25 on T-lymphocytes is down-regulated.

21. A method according to claim 1 or 2 , wherein the T-lymphocytes are derived from metinel lymph nodes draining a metastasis, or they are derived from blood.

22. A method according to claim 2 , wherein the tumour-derived antigen is autologous tumour-derived antigen.

23. A method according to claim 2 , wherein the tumour-derived antigen is autologous denatured tumour extract.

24. A method for treating a patient suffering from a disseminated cancer, the method comprising administering T-lymphocytes to the patient, wherein the T-lymphocytes are CD4+helper or CD8+T-lymphocytes and are obtained by:

i) identifying in a patient one or more metinel lymph nodes, and, optionally, identifying all or part of metastases of the disseminated cancer;

ii) resecting the one or more nodes and, optionally, all or part of the metastases;

iii) isolating metastasis-reactive T-lymphocytes from said lymph nodes; and

iv) in vitro expanding said metastasis-reactive T-lymphocytes by a method comprising:

(a) a first phase of stimulating tumour-reactive CD4+helper or CD8+T-lymphocytes with tumour-derived antigen together with at least one substance having agonistic activity towards the IL-2 receptor to promote survival of tumour-reactive CD4+helper or CD8+T-lymphocytes; and

(b) a second phase of activating and promoting growth of tumour-reactive CD4+helper or CD8+T-lymphocytes, wherein the second phase is initiated when the CD25 cell surface marker or IL-2R marker is down-regulated on T-lymphocytes.