Method for inducing t cells for cell-based immunotherapy from pluripotent stem cells

Patent metadata

Field	Value
Publication number	US-12241089-B2
Application number	US-201515326977-A
Country	US
Kind code	B2
Filing date	Jul 17, 2015
Priority date	Jul 18, 2014
Publication date	Mar 4, 2025
Grant date	Mar 4, 2025

Abstract

Provided is a method for inducing T cells for a cell-based immunotherapy, comprising the steps of:(1) providing Rag 1 and/or Rag 2 gene knockout human pluripotent stem cells bearing genes encoding a T cell receptor specific for a desired antigen, and(2) inducing T cells from the pluripotent stem cells of step (1). Further provided are a cell-based immunotherapy method that uses the T cells for the cell-based immunotherapy and an iPS cell bank for the cell-based immunotherapy.

First claim

What is claimed is:

1. An in vitro method for inducing T cells for a cell-based immunotherapy, where the T cells are CD8 single positive T cells derived from human pluripotent stem cells, comprising the steps of:
(i) providing human pluripotent stem cells having both alleles of Rag-1 or Rag-2 gene knocked out and comprising genes encoding an alpha chain and a beta chain of a T cell receptor specific for a desired antigen,
wherein the human pluripotent stem cells of step (i) are prepared by a method comprising the steps of:
(a) introducing genes encoding the alpha chain and beta chain of the antigen specific T cell receptor for the desired antigen into the pluripotent stem cells; and
(b) knocking out both alleles of the Rag-1 or Rag-2 gene in the human pluripotent stem cells provided in step (a) by genome editing;

(ii) inducing the pluripotent stem cells of step (i) into a population of CD3 positive T cells, wherein the population of CD3 positive T cells comprise CD4CD8 double positive T cells, and CD4CD8 double negative T cells;
(iii) isolating only the CD4CD8 double positive T cells from the population of T cells in step (ii) using flow cytometry; and
(iv) inducing CD8 single positive T cells, which are differentiated from the isolated CD4CD8 double positive T cells of step (iii), and
wherein the CD8 single positive T cells have no rearrangement of the alpha chain of the T cell receptor specific for the desired antigen.

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

3. The method according to claim 1 , wherein the Rag-1 or Rag-2 gene is knocked out by CRISPR-Cas9.

4. The method of claim 2 , wherein the iPS cells have a homozygous HLA haplotype that matches at least one of HLA haplotypes of a subject to be treated.

Assignees

• Univ Kyoto
• Thyas Co Ltd

Inventors

• Kaneko Shin
• Kawamoto Hiroshi
• Masuda Kyoko
• Minagawa Atsutaka
• Hotta Akitsu
• Shimazu Yutaka
• Ichise Hiroshi

Classifications

• A61K40/4243

  Wilms tumor 1 [WT1] · CPC title

• A61K40/46

  Viral antigens · CPC title

• A61K40/32

  T-cell receptors [TCR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• C12N5/0636

  T lymphocytes · CPC title