TY - JOUR
T1 - Technical Advance: Ascorbic acid induces development of double-positive T cells from human hematopoietic stem cells in the absence of stromal cells
AU - Huijskens, Mirelle J. A. J.
AU - Walczak, Mateusz
AU - Koller, Nicole
AU - Briede, Jacob J.
AU - Senden-Gijsbers, Birgit L. M. G.
AU - Schnijderberg, Melanie C.
AU - Bos, Gerard M. J.
AU - Germeraad, Wilfred T. V.
PY - 2014/12
Y1 - 2014/12
N2 - 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.
AB - 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.
KW - T cell precursor
KW - DLL4 protein
KW - adoptive cellular immunotherapy
KW - transplantation
U2 - 10.1189/jlb.1TA0214-121RR
DO - 10.1189/jlb.1TA0214-121RR
M3 - Article
C2 - 25157026
SN - 0741-5400
VL - 96
SP - 1165
EP - 1175
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
IS - 6
ER -