Method of producing vdelta1+ t cells

An in vitro method to generate an expanded population of de novo Notch-induced and differentiated CD1a− Vδ1+ γδ T cells from a cell population comprising human hematopoietic stem/progenitor cells (HPCs) and/or human early thymic progenitors (ETPs), the method comprising a first step of producing a cell composition that comprises a higher amount of γδ T cells than αβ T cells, the first step comprising:

• a. cultivating the cell population comprising the HPCs and/or ETPs in an adequate culture medium comprising Jag2 Notch ligand or agonist thereof;
• and
• b. maintaining the cells in culture for a duration of time sufficient to produce the γδ T cells, wherein the Jag2 Notch ligand or agonist is present or added in an adequate amount to said culture of cells at the time of the culturing and also throughout the culturing period,
• wherein the cell composition resulting from the first step comprises CD1a− Vδ1+ γδ T cells, and wherein the method further comprises a second step of activating and inducing the proliferation of said CD1a− Vδ1+ γδ T cells, the second step comprising cultivating the cells obtained after the first step in an adequate culture medium in the presence of at least one γδTCR agonist and in the presence of at least IL21 and IL15, to obtain a cell population wherein at least 40% of the total γδ T cells are expanded and activated CD1a− Vδ1+ γδ T cells, and wherein said population of CD1a− Vδ1+ γδ T cells wherein:
• at least 40% of the total number of Vδ1+ γδ T cells express CD56 marker,
• at least 60% of the total number of Vδ1+ γδ T cells express NKp44 marker,
• at least 60% of the total number of Vδ1+ γδ T cells express NKp30 marker,
• at least 70% of the total number of Vδ1+ γδ T cells express NKG2D marker, and
• at least 80% of the total number of Vδ1+ γδ T cells express DNAM-1 marker,
• wherein the levels of expression of the above markers are preferably-measured by flow cytometry.