Method for generating T cell progenitors
The invention relates to an in vitro method to generate T cell progenitors, comprising the step of culturing CD34+ cells in a medium containing TNF-alpha and/or an antagonist of the Aryl hydro-carbon/Dioxin receptor, in particular StemRegenin 1 (SR1), in presence of a Notch ligand and optionally a fibronectin fragment.
1. A method for increasing the number of T cells in a human subject in need thereof, comprising administering to said human subject a population of CD7+ T cell progenitors comprising CD7+CD34− cells,
wherein the CD7+CD34− cells represent more than 80% of the population of CD7+ T cell progenitors, and
wherein said population of CD7+ T cell progenitors was prepared by culturing human CD34+ cells in a culture medium comprising fetal serum, TNF-alpha, IL-7, thrombopoietin (TPO), Flt3L, and Stem cell factor (SCF), in the presence of an immobilized Notch ligand and a fibronectin fragment for a period ranging from 5 to 7 days,
wherein said fibronectin fragment comprises the RGDS (SEQ ID NO: 3) and connecting segment-1 (CS-1) motifs as well as a heparin-binding domain,
wherein TNF-alpha is present in the culture medium at a concentration of at least 5 ng/mL,
wherein the Notch ligand is delta-like-ligand 4,
wherein fetal serum is present in the culture medium at a concentration of at least 15%, and
wherein the culture medium does not comprise IL-3.
2. The method of claim 1 , wherein the CD7 + CD34 − cells are also CD5 − .
3. The method of claim 1 , wherein the CD7 + CD34 − cells are also CD1a − .
4. The method of claim 1 , wherein the CD7 + CD34 − cells express a Chimeric Antigen Receptor (CAR).
5. The method of claim 2 , wherein the CD7 + CD34 − CD5 − cells are also CD1a − .
6. The method of claim 1 , wherein the CD34 + cells are isolated from an adult donor.
7. The method of claim 1 , wherein more than 80% of the CD7 + cells are CD34 − CD5 − , CD34 − CD1a − , or CD34 − CD1a − CD5 − .
8. The method of claim 1 , wherein the T cell progenitors comprise a transgene.
9. The method of claim 8 , wherein the transgene is Chimeric Antigen Receptor (CAR).
10. The method of claim 1 , wherein the T cell progenitors are autologous.
11. The method of claim 1 , wherein the T cell progenitors are allogeneic.
12. The method of claim 1 , wherein the T cell progenitors have been conditioned in a pouch prior to administering to the subject.
13. The method of claim 1 , for treating an immunosuppressed human subject.
14. The method of claim 13 , wherein the human subject is immunosuppressed due to a hereditary immune deficiency.
15. The method of claim 13 , wherein the human subject is immunosuppressed due to chemotherapy for leukemia.
16. The method of claim 13 , wherein the human subject is immunosuppressed due to post-graft treatment for prophylaxis of graft-versus-host disease.
17. The method of claim 13 , wherein the human subject is immunosuppressed due to age.
18. The method of claim 13 , wherein the human subject is immunosuppressed due to infection.
19. The method of claim 13 , wherein the human subject is immunosuppressed due to depletion of its immune cells.
20. The method of claim 19 , wherein the human subject is immunosuppressed due to depletion of its immune cells before hematopoietic stem cells transplantation.
21. The method of claim 1 , for treating immunodeficiency.
22. The method of claim 21 , wherein the immunodeficiency is severe combined immunodeficiency (SCID).
23. The method of claim 1 , for treating lymphopenia.
24. The method of claim 1 , for treating cancer.
25. The method of claim 24 , wherein the cancer is leukemia, lymphoma or acute myeloid leukemia.
26. The method of claim 1 , for treating lymphopenia caused or resulting from HIV infection.
27. The method of claim 1 , for treating lymphopenia caused or resulting from partial thymectomy or organ transplant.
28. The method of claim 1 , for treating lymphopenia caused or resulting from autoimmune disease.
29. The method of claim 1 , wherein the medium comprises at least 20% fetal serum.