mRNA expression profile of Lymphocytes by high-throuput sequencing

The activation of CD4+ T helper (Th) cells is crucial for the induction of 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 T-lineage cells (iPS-T cells). Although iPS-T cells retained the same T-cell receptor (TCR) as the original Th clone, their gene expression patterns resembled those of group 1 innate lymphoid cells, and CD4 molecule, an essential co-receptor in TCR-mediated Th responses, was not expressed. The transduction of CD4 genes into iPS-T cells enhanced b3a2 peptide–specific responses. iPS-T cells contained a subpopulation that up-regulates CD40 ligand (CD40L) in response to IL-2 and IL-15. Besides CD4 expression, CD40Lhigh iPS-T cells induced antigen-specific DC maturation characterized by enhanced CD83, CD86, and CCR7. In the presence of Wilms tumor 1 (WT1) peptide, DCs conditioned by CD4-modified CD40Lhigh iPS-T cells stimulated the priming of WT1-specific CTLs. These CTLs eliminated WT1 peptide-expressing CML cell lines in vitro and in vivo. Our findings indicate CD4 modification and purification of CD40Lhigh iPS-T cells generates T helper-like cells that induce effective anti-leukemic CTL responses via DC maturation and suggest feasibility of the cells for adoptive immunotherapy against leukemia. Overall design: Expression profile of the lymphocytes (n = 18) by high-throuput sequencing