title: Technical advance: ascorbic acid induces development of double-positive T cells from human hematopoietic stem cells in the absence of stromal cells.
abstract: 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.
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and mirelle.huijskens@maastrichtuniversity.nl.
authors.forename: Mirelle J A J
authors.initials: MJ
authors.lastname: Huijskens
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Mateusz
authors.initials: M
authors.lastname: Walczak
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Nicole
authors.initials: N
authors.lastname: Koller
authors.affiliation: Department of Toxicogenomics, School for Oncology and Developmental Biology, Maastricht University, Maastricht, the Netherlands.
authors.forename: Jacob J
authors.initials: JJ
authors.lastname: Briedé
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Birgit L M G
authors.initials: BL
authors.lastname: Senden-Gijsbers
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Melanie C
authors.initials: MC
authors.lastname: Schnijderberg
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Gerard M J
authors.initials: GM
authors.lastname: Bos
authors.affiliation: Department of Internal Medicine, Division of Hematology, Maastricht University Medical Center, and.
authors.forename: Wilfred T V
authors.initials: WT
authors.lastname: Germeraad
mesh_terms: D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000096:biosynthesis; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology*; D015704:CD4 Antigens / Q000032:analysis*; D016827:CD8 Antigens / Q000032:analysis*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002956:Citrulline / Q000378:metabolism; D018920:Coculture Techniques; D000069585:Filgrastim; D020869:Gene Expression Profiling; D015329:Gene Rearrangement, T-Lymphocyte; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D018189:Immunomagnetic Separation; D016023:Integrins / Q000032:analysis; D038041:Lymphopoiesis / Q000187:drug effects*; D009569:Nitric Oxide / Q000378:metabolism; D061251:Primary Cell Culture / Q000379:methods; D019707:Receptors, Chemokine / Q000032:analysis; D011994:Recombinant Proteins / Q000494:pharmacology; D017154:Stromal Cells; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D000945:Antigens, Differentiation, T-Lymphocyte; D000975:Antioxidants; D015704:CD4 Antigens; D016827:CD8 Antigens; D016023:Integrins; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D019323:omega-N-Methylarginine; D002956:Citrulline; D009569:Nitric Oxide; D001205:Ascorbic Acid; D000069585:Filgrastim
keywords: DLL4 protein; T cell precursor; adoptive cellular immunotherapy; transplantation