Method for producing helper T cells from pluripotent stem cells
A method of producing helper T cells, comprising: (i) culturing T cells, which have been induced from pluripotent stem cells and into which a CD4 gene or a gene product thereof has been introduced, in a medium containing IL-2 and IL-15; and (ii) isolating CD40L-highly expressing T cells from cells obtained in step (i).
1. A method of producing helper T cells comprising:
(i) culturing T cells, which have been induced from pluripotent stem cells and into which a CD4 gene or a gene product thereof has been introduced, in a medium containing IL-2 and IL-15; and
(ii) isolating CD40L-highly expressing T cells from cells obtained in Step (i).
2. The method according to claim 1 , wherein the concentration of the IL-2 is 10 to 500 IU/ml, and the concentration of the IL-15 is 1 to 50 ng/ml.
3. The method according to claim 1 , wherein the T cells have been induced from the pluripotent stem cells by a method comprising:
(1) inducing CD34-positive hematopoietic progenitor cells from pluripotent stem cells; and
(2) culturing CD34-positive hematopoietic progenitor cells obtained in Step (1), in the presence of FLT3L and IL-7.
4. The method according to claim 3 , wherein Step (1) comprises co-culturing pluripotent stem cells with C3H10T1/2, followed by co-culturing with C3H10T1/2 in the presence of VEGF, FLT3L, and SCF.
5. The method according to claim 3 , wherein Step (2) comprises co-culturing the CD34-positive hematopoietic progenitor cells with stromal cells.
6. The method according to claim 3 , wherein said method of inducing the T cells from the pluripotent stem cells further comprises:
(3) co-culturing cells obtained in Step (2), with peripheral blood mononuclear cells in the presence of IL-7 and IL-15.
7. The method according to claim 3 , wherein said method of inducing T cells from the pluripotent stem cells further comprises:
bringing cells obtained in Step (2) into contact with mitogen, and/or
bringing cells obtained in Step (3) into contact with mitogen.
8. The method according to claim 1 , wherein the CD4 gene has been introduced using a retrovirus vector.
9. The method according to claim 1 , wherein the pluripotent stem cells are pluripotent stem cells having a rearranged TCR sequence of interest.
10. The method according to claim 9 , wherein the pluripotent stem cells are human iPS cells induced from lymphocytes that recognize a desired antigen(s).
11. The method according to claim 10 , wherein the lymphocytes that recognize a desired antigen(s) are lymphocytes that recognize BCR/ABL.