Ross D. Jones, Kevin Salim, Laura N. Stankiewicz, John M. Edgar, Lorna Leon, Jana K. Gillies, Ali Murtaza, Lauren J. Durland, Divy Raval, Charles Lau, Thristan Paulo B. Taberna, Han Hsuan Hsu, Carla Zimmerman, Yale S. Michaels, Fábio Rossi, Megan K. Levings, Peter W. Zandstra
Allogeneic T cell therapies are a highly desirable option to circumvent the cost and complexity of using autologous T cells to treat diseases. Allogeneic CD8 + T cells can be made from pluripotent stem cells (PSCs), but deriving CD4 + T cells from PSCs has remained a significant challenge. Using feeder- and serum-free conditions, we found that CD4 + vs. CD8 + T cell commitment from PSCs can be controlled by fine-tuning the dynamics of Notch and T cell receptor (TCR) signaling delivered to CD4 + CD8 + double-positive T cells. Notch signaling negatively impacts CD4 + T cell commitment, and its timed removal allows generation of clonally diverse and expandable CD4 + T cells from PSCs. The resulting CD4 + T cells respond to cytokine-mediated polarization by differentiating into Th1, Th2, or Th17 cells, recapitulating canonical helper cell function. These findings represent a significant step toward using PSC-derived CD4 + T cells as a low-cost, off-the-shelf cell therapy.