CD34+CD7+CD5+CD1a-human progenitor T-cells produced in vitro and methods of using
Human progenitor T cells that are able to successfully engraft a murine thymus and differentiate into mature human T and NK cells are described The human progenitor T cells have the phenotype CD34+CD7+CD1a−CD5− or CD34+CD7+CD1a−CD5+ and are derived from human hematopoietic stem cells, embryonic stem cells and induced pluripotent stem cells b\ coculture with cells expressing a Notch receptor ligand (OP9-DL1 or OP9-DL4) Such cells are useful in a variety of applications including immune reconstitution, the treatment of immunodeficiencies and as carriers for genes used in gene therapy.
1. A method of producing a CD34 + CD7 + CDS + CD1a − progenitor T cell (pro T cell) comprising
(a) culturing a mammalian hematopoietic stem cell (HSC) with a stromal cell that express a Notch ligand in a medium comprising Flt-3L and IL-7; and
(b) determining the presence of CD34 + CD7 + CD5 + CD1a − pro-T cells in the culture.
2. The method of claim 1 wherein the HSC is a human HSC.
3. The method according to claim 1 wherein the Notch ligand is DL1 or DL4.
4. The method of claim 3 wherein the cell expressing the Notch ligand is an OP-9 cell.
5. A method for increasing the number of T cells in a mammal comprising administering to a mammal an effective amount of a purified human progenitor T cell having a phenotype CD34 + CD7 + CD5 + CD1a − , wherein the human progenitor T cell is isolated according to the method of claim 1 , wherein an increase in CD7 + CD5 + CD1a + pro-T cells and cells of a later T-cell developmental stage is seen.
6. The method according to claim 5 wherein the mammal has cancer.
7. The method according to claim 5 wherein the mammal has HIV/AIDS.
8. The method according to claim 5 wherein the mammal has an autoimmune disease.
9. The method according to claim 5 wherein the progenitor T cell contains a heterologous gene.