IP Library › Granted Patent US 12,391,921
Granted Patent B2
US 12,391,921 · App. 17/724,907 · Granted Aug 19, 2025

Method for inducing antigen specific CD8 positive T cells

Inventors: Hiroshi Kawamoto (Kyoto, JP); Takuya Maeda (Kyoto, JP); Kyoko Masuda (Kyoto, JP)
Assignee: KYOTO UNIVERSITY
C12N5/0636A61K40/11A61K40/32A61K40/4243C12N5/10A61K2239/31A61K2239/38A61K2239/48C12N2501/48C12N2501/998C12N2506/45
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,391,921
App. No.
17/724,907
Granted
Aug 19, 2025
Kind
B2
Abstract

Provided is a method for inducing CD4 − CD8 + T cells having an antigen specific cytotoxic activity from pluripotent stem cells, comprising the steps of: (1) differentiating pluripotent stem cells to give a cell culture comprising CD4 − CD8 − T cells and CD4 + CD8 + T cells, (2) removing CD4 − CD8 − cells from the cell culture obtained in step (1), and (3) differentiating the CD4 + CD8 + cells in the cell culture into CD4 − CD8 + T cells.

Claims (21)

1. A method for inducing CD4 − CD8 + T cells, comprising the steps of:

(1) differentiating pluripotent stem cells to give a cell culture comprising CD4 − CD8 + T cells and CD4 − CD8 + T cells,

(2) differentiating the CD4 − CD8 + T cells in the cell culture obtained in (1) into CD4 − CD8 + T cells under the presence of a substance that inhibits cytotoxic activity of CD4 − CD8 + cells.

2. The method according to claim 1 , wherein the substance that inhibits cytotoxic activity of CD4 − CD8 + T cells is selected from the group consisting of a perforin inhibitor, a granzyme inhibitor, a Fas pathway inhibitor, a caspase inhibitor, and an inhibitory antibody against Natural Killer activating receptor.

3. The method according to claim 1 , wherein the pluripotent stem cells are iPS cells.

4. The method according to claim 3 , wherein the pluripotent stem cells are iPS cells homozygous for HLA haplotype.

5. The method according to claim 1 , wherein the pluripotent stem cells comprise rearranged genes encoding a T cell receptor or a chimeric antigen receptor specific for an antigen and CD4 − CD8 + T cells having a cytotoxic activity specific for the antigen are induced.

6. The method according to claim 5 , wherein the pluripotent stem cells comprise rearranged genes encoding a T cell receptor specific for the antigen.

7. The method according to claim 6 , wherein the pluripotent stem cells are T-iPS cells induced from human T cells comprising rearranged genes encoding a T cell receptor specific for the antigen.

8. The method according to claim 6 , wherein the pluripotent stem cells are TCR-iPS cells obtained by introducing rearranged genes encoding a T cell receptor specific for the antigen into human T cells.

9. The method according to claim 5 , wherein the step of differentiating CD4 − CD8 + cells into CD4 − CD8 + T cells is performed by directly activating any one of the activation pathways which occur upon stimulation of the T cell receptor.

10. The method according to claim 9 , wherein the step of differentiating CD4 − CD8 + cells into CD4 − CD8 + T cells is performed by using anti-CD3 antibody.

11. The method according to claim 9 , wherein the step of differentiating CD4 − CD8 + cells into CD4 − CD8 + T cells is performed by stimulating the cells with the antigen.

12. The method according to claim 11 , wherein the step of differentiating CD4 − CD8 + cells into CD4 − CD8 + T cells is performed by stimulating the cells with antigen presenting cells that present the antigen.

13. The method according to claim 1 , for establishment of CD4 − CD8 + T cells having a cytotoxic activity specific for the antigen, further comprising the step of introducing a T cell receptor or a chimeric antigen receptor specific for the antigen into the induced CD4 − CD8 + T cells.

14. The method according to claim 13 , for introducing a T cell receptor specific for the antigen into the induced CD4 − CD8 + T cells.

15. The method according to claim 5 , further comprising the step of further proliferating the obtained CD4 − CD8 + T cells specific for the antigen.

16. The method according to 15 , wherein the step of further proliferating the obtained CD4 − CD8 + T cells is performed by directly activating any part of the activation pathway which occurs on stimulating a T cell receptor.

17. The method according to claim 15 , wherein the step of further proliferating the obtained CD4 − CD8 + T cells is performed by using anti-CD3 antibody.

18. The method according to claim 15 , wherein the step of further proliferating the obtained CD4 − CD8 + T cells is performed by stimulating the cells with the antigen.

19. The method according to claim 18 , wherein the step of further proliferating the obtained CD4 − CD8 + T cells is performed by stimulating the cells with antigen presenting cells that present the antigen.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 20, 2022
From: KAWAMOTO, HIROSHI; MAEDA, TAKUYA; MASUDA, KYOKO
To: KYOTO UNIVERSITY
Reel/Frame 059651/0010 →
Priority Claims (1)
JP 2016-082410 · Apr 15, 2016 · national
Continuity (2)
Continuation 16092411
Related Publication 20220251506A1 · Aug 11, 2022
References Cited (116)
US 20080213885A1 · Tryggvason et al. · 2008 [cited by applicant]
US 20090047263A1 · Yamanaka et al. · 2009 [cited by applicant]
US 20090068742A1 · Yamanaka et al. · 2009 [cited by applicant]
US 20100075241A1 · Caimi et al. · 2010 [cited by applicant]
US 20110117645A1 · Yasuda et al. · 2011 [cited by applicant]
US 20110287538A1 · Fusaki et al. · 2011 [cited by applicant]
US 20120220031A1 · Sekiguchi et al. · 2012 [cited by applicant]
US 20130078226A1 · Nakauchi et al. · 2013 [cited by applicant]
US 20170128556A1 · Kawamoto et al. · 2017 [cited by applicant]
US 20170296649A1 · Kawamoto et al. · 2017 [cited by applicant]
US 20170326175A1 · Kaneko et al. · 2017 [cited by applicant]
US 20170369850A1 · Kaneko et al. · 2017 [cited by applicant]
EP 2853590A1 · 2015 [cited by applicant]
JP 2014514927A · 2014 [cited by applicant]
WO WO2007069666A1 · 2007 [cited by applicant]
WO WO2008118820A2 · 2008 [cited by applicant]
WO WO2009007852A2 · 2009 [cited by applicant]
WO WO2009032194A1 · 2009 [cited by applicant]
WO WO2009057831A1 · 2009 [cited by applicant]
WO WO2009058413A1 · 2009 [cited by applicant]
WO WO2009079007A1 · 2009 [cited by applicant]
WO WO20090775119A1 · 2009 [cited by applicant]
WO WO2009091659A2 · 2009 [cited by applicant]
WO WO2009101084A1 · 2009 [cited by applicant]
WO WO2009101407A2 · 2009 [cited by applicant]
WO WO2009102983A2 · 2009 [cited by applicant]
WO WO2009114949A1 · 2009 [cited by applicant]
WO WO2009117439A2 · 2009 [cited by applicant]
WO WO2009123349A1 · 2009 [cited by applicant]
WO WO2009126250A2 · 2009 [cited by applicant]
WO WO2009126251A2 · 2009 [cited by applicant]
WO WO2009126655A2 · 2009 [cited by applicant]
WO WO2009157593A1 · 2009 [cited by applicant]
WO WO2010008054A1 · 2010 [cited by applicant]
WO WO2010009015A2 · 2010 [cited by applicant]
WO WO2010033906A2 · 2010 [cited by applicant]
WO WO2010033920A2 · 2010 [cited by applicant]
WO WO2010042800A1 · 2010 [cited by applicant]
WO WO2010050626A1 · 2010 [cited by applicant]
WO WO2010056831A2 · 2010 [cited by applicant]
WO WO2010068955A2 · 2010 [cited by applicant]
WO WO2010098419A1 · 2010 [cited by applicant]
WO WO2010102267A2 · 2010 [cited by applicant]
WO WO2010111409A2 · 2010 [cited by applicant]
WO WO2010111422A2 · 2010 [cited by applicant]
WO WO2010115050A2 · 2010 [cited by applicant]
WO WO2010124290A2 · 2010 [cited by applicant]
WO WO2010137746A1 · 2010 [cited by applicant]
WO WO2010147395A2 · 2010 [cited by applicant]
WO WO2010147612A1 · 2010 [cited by applicant]
WO WO2011043405A1 · 2011 [cited by applicant]
WO WO2011096482A1 · 2011 [cited by applicant]
WO WO2013176197A1 · 2013 [cited by applicant]
WO WO2015099134A1 · 2015 [cited by applicant]
WO WO2016010154A1 · 2016 [cited by applicant]
WO WO201610153A1 · 2016 [cited by applicant]
WO WO2016010148A1 · 2016 [cited by applicant]
WO WO2016076415A1 · 2016 [cited by applicant]
Osinska et al. (2014, Cenr. Eur. J Immunol., vol. 39(1), pp. 109-115.) (Year: 2014). [cited by examiner]
Alberts et al., Molecular Biology of the Cell. 4th edition. New York: Garland Science; 2002. The Generation of Antibody Diversity. Available from: https://www.ncbi.nlrmnih.gov/books/N BK26860/ (Year: 2002). [cited by applicant]
Ando et al., “A safeguard system for induced pluripotent stem cell-derived rejuvenated T cell therapy,” [cited by applicant]
Arstila et al., A direct estimate of the human alphabeta T cell receptor diversity. Science. Oct. 29, 1999;286(5441):958-61. doi: 10.1126/science.286.5441.958. PMID:10542151. [cited by applicant]
Chen et al., “The role and mechanisms of double negative regulatory T cells in the suppression of immune responses”, [cited by applicant]
Cheroutre et al. “Doubting the TCR coreceptor function of CD8alphaalpha”, [cited by applicant]
Cyranoski et al., “Stem-cell pioneer banks on future therapies”, [cited by applicant]
Eminli et al., “Reprogramming of neural progenitor cells into induced pluripotent stem cells in the absence of exogenous Sox2,” [cited by applicant]
Feng et al., “Reprogramming of fibroblasts into induced pluripotent stem cells with orphan nuclear receptor Esrrb”, [cited by applicant]
Grskovic et al., “Induced pluripotent stem cells-opportunities for disease modelling and drug discovery,” [cited by applicant]
Han et al., “Tbx3 improves the gem-line competency of induced pluripotent stem cells”, [cited by applicant]
Heng et al., “The nuclear receptor Nr5a2 can replace Oct4 in the reprogramming of murine somatic cells to pluripotent cells”, [cited by applicant]
Huangfu et al., “Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2”, [cited by applicant]
Huangfu et al., “Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds”, [cited by applicant]
Ichida et al., “A small-molecule inhibitor of Tgf-beta signaling replaces Sox2 in reprogramming by inducing Nanog”, [cited by applicant]
International Search Report for International Application No. PCT/JP2017/015358, mailing date Jul. 11, 2017, 2 pages. English translation. [cited by applicant]
International Preliminary Report on Patentability for International Patent Application No. PCT/JP2017/015358, dated Jan. 29, 2018. [cited by applicant]
Judson et al., “Embryonic stem cell-specific microRNAs promote induced pluripotency”, [cited by applicant]
Kaneko S., “In vitro generation of rejuvenated antigen-specific T cells by reprogramming to pluripotency and redifferentiation”, [cited by applicant]
Kawamoto H. et al., “Cloning and expansion of antigen-specific T cells using iPSC technology: A novel strategy for cancer immunotherapy”, [cited by applicant]
Kim et al., “Direct reprogramming of human neural stem cells by OCT4”, [cited by applicant]
Kitayama et al., “Cellular adjuvant properties, direct cytotoxicity of re-differentiated V-alpha24 invariant NKT-ells from human induced pluripotent stem cells”, [cited by applicant]
Lyssiotis et al., “Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical ,implementation of Klf4”, [cited by applicant]
Maeda et al., “Regeneration of CD8αβ T cells from T-cell-derived iPSC imparts potent tumor antigen-specific cytoxicity”, [cited by applicant]
Maekawa et al., “Direct reprogramming of somatic cells is promoted by matemal transcription factor Glis1”, [cited by applicant]
Mali et al., “Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes”, [cited by applicant]
Marson et al., “Wnt Signalling Promotes Reprogramming of Somatic Cells to Pluripotency”, [cited by applicant]
Nakagawa et al., “Generation of induced pluripotent stem cells without Myc from mouse and human Fibroblasts”, [cited by applicant]
Nakao et al., “Expansion of functional human mucosal-associated invariant T cells via reprogramming to pluripotency and redifferentiation”, [cited by applicant]
Narren et al., “Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA”, [cited by applicant]
Neko S., “In vitro generation of antigen-specific T cells from induced pluripotent stem cells of antigen-specific T cell origin”, [cited by applicant]
Newell et al., “Beyond model antigens: high-dimensional methods for the analysis of antigen-specific T cells”, [cited by applicant]
Nishimura et al., “Development of detective and persistent sendai virus vector,” [cited by applicant]
Nishimura et al., “Generation of Rejuvenated Antigen-Specific T Cells by Reprogramming to Pluripotency and Redifferentiation”, [cited by applicant]
Saito et al., “Reprogramming of Melanoma Tumor-Infiltrating Lymphocytes to Induced Pluripotent Stem Cells”, [cited by applicant]
Saito et al., “Adoptive Transfer of CD8+ T Cells Generated from Induced Pluripotent Stem Cells Triggers Regressions of Large Tumors Along with Immunological Memory”, [cited by applicant]
Schmitt et al. “New Strategies in Engineering T-cell Receptor Gene-Modified T cells to More Effectively Target Malignancies”, [cited by applicant]
Shi et al., “A combined chemical and genetic approach for the generation of induced pluripotent stem cells”, [cited by applicant]
Shi et al., “Induction of pluripotent stem cells from mouse embryonic fibroblasts by Oct4 and Klf4 with small-molecule compounds”, [cited by applicant]
Stadtfeld et al., “Induced pluripotent stem cells generated without viral integration”, [cited by applicant]
Su et al., “Memory T Cell From Granzyme B Attack by Double-Negative T Regulatory Cell”, [cited by applicant]
Sun et al., “Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells”, [cited by applicant]
Supplementary European Search Report in related European Application No. 17782533.8, dated Nov. 19, 2020 8 pages. [cited by applicant]
Takahashi et al., “Induction of pluripotent stem cells from mouse embryonic and ault fibroblasts cultures by lamed factors”, [cited by applicant]
Takahashi et al., “Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors,” [cited by applicant]
Takahashi et al., “Human induced pluripotent stem cells on autologous feeders,” PLoS One, 4:e8067, 2009. [cited by applicant]
Takuya M. et al., iPS Saibo Gijutsu o Mochiita WT1 Tokuiteki Killer T-saibo (CTL) no Saisei, Regenerative Medicine, vol. 15, Suppl., p. 258, 2016. [cited by applicant]
Themeli et al., “Generation of tumor-targeted human T lymphocytes from induced pluripotent stem cells for cancer therapy”, [cited by applicant]
Themeli et al., “New cell sources for T cell engineering and adoptive immunotherapy,” [cited by applicant]
Timmermans et al., “Generation of T Cells from Human Embryonic Stem Cell-Derived Hematopoietic Zones”, [cited by applicant]
Tsuboi et al., “Enhanced induction of human WT1-specific cytotoxic T lymphocytes with a 9-mer WT1 peptide modified at HLA-A*2402-binding residues,” [cited by applicant]
Yu et al., “Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells”, [cited by applicant]
Vizcardo et al., “Regeneration of human tumor antigen-specific T cells from iPSCs derived from mature CD8+ T Cells”, [cited by applicant]
Zhao et al., “Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation,” [cited by applicant]
Cyranoski et al., “Stem-cell pioneer banks on future therapies”, Nature, Aug. 9, 2012, 488: 139. [cited by applicant]
Kaneko, “In Vitro Generation of Antigen-Specific T Cells from Induced Pluripotent Stem Cells of Antigen-Specific T Cell Origin”, Tumor Immunology Methods and Protocols, Chapter 6, Apr. 1, 2016, Part of the Methods in Mo… [cited by applicant]
Wakao et al., “Expansion of functional human mucosal-associated invariant T cells via reprogramming to pluripotency and redifferentiation”, Cell Stem Cell, May 2, 2013, 12: 546-558. [cited by applicant]
Warren et al., “Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA”, Cell Stem Cell, Nov. 5, 2010, 7: 618-630. [cited by applicant]