Method for producing helper T cells from pluripotent stem cells

Patent metadata

Field	Value
Publication number	US-11559548-B2
Application number	US-201816494265-A
Country	US
Kind code	B2
Filing date	Mar 13, 2018
Priority date	Mar 14, 2017
Publication date	Jan 24, 2023
Grant date	Jan 24, 2023

Abstract

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).

First claim

What is claimed is:

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.

Assignees

• Univ Kyoto

Inventors

• Kaneko Shin
• Ueda Norihiro
• Uemura Yasushi
• Fukuda Kyoko

Classifications

• C12N2501/2315

  Interleukin-15 (IL-15) · CPC title

• C12N2502/1394

  Bone marrow stromal cells; whole marrow · CPC title

• C12N15/86 Primary

  Viral vectors · CPC title

• C12N2501/999

  Small molecules not provided for elsewhere · CPC title

• A61K35/17 Primary

  Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) · CPC title