Methods for obtaining regulatory T cells and uses thereof
Disclosed is a method for obtaining a population of human Treg cells including the steps of: (a) culturing a population of human monocytes with a medium including an amount of an interleukin-34 (IL-34) polypeptide in order to obtain a population of immunosuppressive macrophages; (b) co-culturing a population of human peripheral blood mononuclear cells (PBMCs) and the population of immunosuppressive macrophages obtained at step (a).
1. A method for obtaining a population of human CD8+ Treg cells comprising the steps of: (a) culturing a population of human monocytes in presence of interleukin-34 (IL-34) polypeptide in order to obtain a population of immunosuppressive macrophages; (b) co-culturing a population of human peripheral blood mononuclear cells (PBMCs) and the population of immunosuppressive macrophages obtained at step (a) to obtain a population of human CD8+ Treg; and (c) optionally isolating the human CD8+ Treg cells from the population obtained at step (b).
2. The method of claim 1 , wherein the population of immunosuppressive macrophages are allogeneic to the population of human PBMCs.
3. The method of claim 1 , wherein the population of human monocytes is a population of CD14+ human monocytes.
4. The method of claim 1 , wherein the IL-34 polypeptide is human IL-34 polypeptide.
5. The method of claim 1 , wherein the IL-34 polypeptide comprises or consists of SEQ ID NO: 1.
6. The method of claim 1 , wherein the IL-34 polypeptide is added to the culture at a concentration ranging from 1 to 500 ng/ml.
7. The method of claim 6 , wherein the IL-34 polypeptide is added to the culture at a concentration of 50 ng/ml.
8. The method of claim 1 , wherein the co-culture of PBMCs and the population of immunosuppressive macrophages obtained at step (a) is performed in a medium comprising IL-15 and/or IL-2.
9. The method of claim 8 , wherein the cytokines are human IL-15 and/or human IL-2.
10. The method of claim 1 , wherein the Treg cells are CD8+Foxp3+ Treg cells.
11. The method of claim 1 , wherein the Treg cells are CD8+CD45RClow Treg cells.
12. The method of claim 1 , wherein the Treg cells are CD8+Foxp3+CD45RClow Treg cells.