US 11,987,811 B2
Method for producing CD4-positive T cells from pluripotent stem cells
Shin Kaneko, Kyoto (JP); Norihiro Ueda, Kyoto (JP); and Yasushi Uemura, Kashiwa (JP)
Assigned to KYOTO UNIVERSITY, Kyoto (JP)
Appl. No. 15/767,917
Filed by Kyoto University, Kyoto (JP)
PCT Filed Oct. 14, 2016, PCT No. PCT/JP2016/080582
§ 371(c)(1), (2) Date Apr. 12, 2018,
PCT Pub. No. WO2017/065288, PCT Pub. Date Apr. 20, 2017.
Claims priority of application No. 2015-203482 (JP), filed on Oct. 15, 2015.
Prior Publication US 2018/0298337 A1, Oct. 18, 2018
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0783 (2010.01); A61K 35/17 (2015.01); C12N 5/0735 (2010.01); C12N 15/86 (2006.01); A61K 39/00 (2006.01); A61P 35/02 (2006.01)
CPC C12N 5/0637 (2013.01) [A61K 35/17 (2013.01); C12N 5/0606 (2013.01); C12N 15/86 (2013.01); A61K 39/00 (2013.01); A61P 35/02 (2018.01); C12N 2500/30 (2013.01); C12N 2501/125 (2013.01); C12N 2501/165 (2013.01); C12N 2501/26 (2013.01); C12N 2502/11 (2013.01); C12N 2502/1171 (2013.01); C12N 2502/1323 (2013.01); C12N 2506/45 (2013.01); C12N 2510/00 (2013.01); C12N 2740/10043 (2013.01); C12N 2770/00043 (2013.01)] 9 Claims
 
1. An in vitro method for inducing CD4-positive T cells, the method comprising:
(a) providing T-cells comprising a reconstituted T cell receptor (TCR) gene induced from an induced pluripotent stem cell, by a process comprising:
(i) reprogramming a T helper cell comprising the TCR gene and recognizing an antigen in a culture medium to produce an induced pluripotent stem (iPS) cell comprising the reconstituted TCR gene;
(ii) inducing CD34-positive hematopoietic progenitor cells from the pluripotent stem cells;
(iii) culturing the CD34-positive hematopoietic progenitor cells of step (ii) in the presence of FLT3L and IL-7; and
(iv) co-culturing the cells obtained from step (iii) with peripheral blood nuclear cells in presence of a mitogen, IL-7 and IL-15 to produce T cells comprising the reconstituted TCR gene, and
(b) introducing a CD4 gene or a CD4 gene product into the T cells such that CD4-positive T cells comprising the reconstituted TCR gene are obtained,
wherein the CD4-positive T-cells exhibit growth in response to the antigen stimulation, produce interferon-γ (IFN-γ) in response to the antigen stimulation, and activate dendritic cells (DCs), and wherein said induced pluripotent stem cells comprise the reconstituted TCR gene.