Technical Advance: Ascorbic acid induces development of double-positive T cells from human hematopoietic stem cells in the absence of stromal cells

Mirelle J. A. J. Huijskens*, Mateusz Walczak, Nicole Koller, Jacob J. Briede, Birgit L. M. G. Senden-Gijsbers, Melanie C. Schnijderberg, Gerard M. J. Bos, Wilfred T. V. Germeraad

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

53 Citations (Web of Science)

Abstract

In vitro human T cell development from hematopoietic stem cells in the presence and absence of feeder cells is promoted by ascorbic acid. The efficacy of donor HSCT is partly reduced as a result of slow post-transplantation immune recovery. In particular, T cell regeneration is generally delayed, resulting in high infection-related mortality in the first years post-transplantation. Adoptive transfer of in vitro-generated human T cell progenitors seems a promising approach to accelerate T cell recovery in immunocompromised patients. AA may enhance T cell proliferation and differentiation in a controlled, feeder-free environment containing Notch ligands and defined growth factors. Our experiments show a pivotal role for AA during human in vitro T cell development. The blocking of NOS diminished this effect, indicating a role for the citrulline/NO cycle. AA promotes the transition of proT1 to proT2 cells and of preT to DP T cells. Furthermore, the addition of AA to feeder cocultures resulted in development of DP and SP T cells, whereas without AA, a preT cell-stage arrest occurred. We conclude that neither DLL4-expressing feeder cells nor feeder cell conditioned media are required for generating DP T cells from CB and G-CSF-mobilized HSCs and that generation and proliferation of proT and DP T cells are greatly improved by AA. This technology could potentially be used to generate T cell progenitors for adoptive therapy.
Original languageEnglish
Pages (from-to)1165-1175
JournalJournal of Leukocyte Biology
Volume96
Issue number6
DOIs
Publication statusPublished - Dec 2014

Keywords

  • T cell precursor
  • DLL4 protein
  • adoptive cellular immunotherapy
  • transplantation

Cite this