Generation of TCR-Expressing Innate Lymphoid-like Helper Cells that Induce Cytotoxic T Cell-Mediated Anti-leukemic Cell Response

Abstract

SummaryCD4+ T helper (Th) cell activation is essential for inducing cytotoxic T lymphocyte (CTL) responses against malignancy. We reprogrammed a Th clone specific for chronic myelogenous leukemia (CML)-derived b3a2 peptide to pluripotency and re-differentiated the cells into original TCR-expressing T-lineage cells (iPS-T cells) with gene expression patterns resembling those of group 1 innate lymphoid cells. CD4 gene transduction into iPS-T cells enhanced b3a2 peptide-specific responses via b3a2 peptide-specific TCR. iPS-T cells upregulated CD40 ligand (CD40L) expression in response to interleukin-2 and interleukin-15. In the presence of Wilms tumor 1 (WT1) peptide, antigen-specific dendritic cells (DCs) conditioned by CD4-modified CD40Lhigh iPS-T cells stimulated WT1-specific CTL priming, which eliminated WT1 peptide-expressing CML cells in vitro and in vivo. Thus, CD4 modification of CD40Lhigh iPS-T cells generates innate lymphoid helper-like cells inducing bcr-abl-specific TCR signaling that mediates effectiveanti-leukemic CTL responses via DC maturation, showing potential for adjuvant immunotherapy against leukemia.

Toward the development of true “off‐the‐shelf” synthetic T‐cell immunotherapy

“…Although the aforementioned studies show the potential of iPSCderived T cells as an alternative cell source for T cell immunotherapy, recent studies, including those at our laboratory, revealed that T cells differentiated from iPSCs using the current differentiation methods display features similar to γδT cells or innate lymphoid cells. [30][31][32] Current differentiation culture induces T cells expressing CD56, a marker for natural killer cells, during multiple rounds of expansion. General schema illustrating autologous T cell manufacturing.…”

A clinically applicable and scalable method to regenerate T-cells from iPSCs for off-the-shelf T-cell immunotherapy

“…Previous studies have shown that HECs or HPCs differentiated from ESCs or iPSCs could be induced to differentiate into the T-cell lineage by co-culturing with OP9 feeder layer-expressing notch ligands, DL1 or DL4, with a limited efficiency of αβ- or γδ-TCR + cell generation 2 , 3 , 32 . It has been also demonstrated that generation of iPSC from T-cells (T-iPSCs) and redifferentiation enhance the efficiency of αβTCR-expressing cell generation from iPSCs 4 – 7 , 33 , 34 . Recently, we have also shown that exogenous TCR transduction in clinical-grade iPSCs can be used as a starting material to generate antigen-specific TCR-engineered iCD8αβ + T-cells with in vivo activity, thereby minimizing inter-clonal variations of iPSCs and bypassing laborious iPSC generation processes 9 .…”

“…PBMC samples were obtained from healthy volunteers who provided written informed consent. The H25-4, H25-31, and GPC3#16-1 iPSC clones were generated from CTL clones by Sendai virus reprogramming vectors as previously described 4 , 9 , 33 , 34 . An HLA homozygous iPSC line FfI01s04 was established in CiRA and distributed with informed consent and permission of the Ethical Review Board at CiRA.…”

Type I Interferon Delivery by iPSC-Derived Myeloid Cells Elicits Antitumor Immunity via XCR1+ Dendritic Cells

“…iPSCs represent a potential immune cell source for cancer immunotherapy (Sachamitr et al, 2014). However, despite their ability to differentiate into various somatic cell types, a largescale cell culture system, extensive labor, and long periods for cell differentiation are required to generate sufficient immune cells from iPSCs (Kitayama et al, 2016;Senju et al, 2009;Ueda et al, 2018). We previously developed a simple and efficient method for obtaining numerous myeloid cells from human and mouse iPSCs (Haruta et al, 2013;Zhang et al, 2015).…”