Generation of effector CD4⁺ T cells from human iPSC

Off-the-shelf T cell therapies could be transformative in chimeric antigen receptor (CAR) therapies for cancers and treatment of chronic inflammatory diseases. However, challenges remain in generating CD4⁺ T cells from induced pluripotent stem cells (iPSCs). We describe a key role for the withdrawal of Notch ligand during T cell receptor stimulation of CD4/CD8 double-positive progenitors allowing access to the CD4⁺ lineage in iPSC T cells (iCD4⁺ T cells). Functional analyses of iCD4⁺ T cells by using a novel high-parameter cytometry by time-of-flight (CyTOF) intracellular cytokine panel revealed canonical Th1 cytokine signatures and cells producing varying combinations of other cytokines, including IL-4, IL-8, and IL-13. Single-cell RNA sequencing of iCD4⁺ T cells demonstrated a transcriptional signature similar to human peripheral CD4⁺ T cells. We believe this robust yet simple platform represents a key step toward off-the-shelf iCD4⁺ T cell therapies with utility for the treatment of a panoply of diseases including cancer and inflammatory autoimmune disorders.