Generation of T cells from adult human hematopoietic stem cells and progenitors in a fetal thymic organ culture system: stimulation by tumor necrosis factor-α

Abstract

To investigate the T-lymphopoietic capacity of human adult bone marrow (ABM) hematopoietic progenitor cells, CD34+Lin−, CD34+CD38+, and CD34++CD38− cells were cultured in a severe combined immunodeficient (SCID) mouse fetal thymic organ culture (FTOC). Direct seeding of these progenitors resulted in a moderate to severe cell loss, particularly for the CD34++CD38− cell fraction, and T cells could only be generated from the CD34+Lin− fraction. Preincubation for 36 hours with interleukin-3 (IL-3) and stem cell factor (SCF) led to an improved cell survival and proliferation, although T-cell development was seen only in the CD34+Lin− fraction. Addition of tumor necrosis factor (TNF)- to IL-3 + SCF-supplemented preincubation medium resulted in optimal cell survival, cell proliferation. and T-cell generation of all 3 cell fractions. The TNF- effect resulted in an up-regulation of CD127 (ie, the IL-7 receptor -chain) in a small subset of the CD34+ cells. No evidence could be generated to support the possibility that TNF- inhibits a cell population that suppresses T-cell differentiation. A quantitatively different T-cell generation potency was still seen between the 3 subpopulations: CD34+Lin− (100% success rate) > CD34+CD38+ (66%) > CD34++CD38− (25%). These data contrast with our previous findings using fetal liver and cord blood progenitors, which readily differentiate into T-lymphocytes in FTOC, even without prestimulation with cytokines. Our results demonstrate that adult CD34++CD38− cells, known to contain hematopoietic stem cells, can differentiate into T-lymphocytes and that a significant difference exists in T-lymphopoietic activity of stem cells derived from ontogenetically different sources.

Tumor Necrosis Factor Promotes Human T‐Cell Development in Nonobese Diabetic/Severe Combined Immunodeficient Mice

“…Zuniga‐Pflucker et al [24] have shown that TNF is important for the induction of critical events leading to murine T‐lineage commitment and maturation in a thymus reconstitution assay, acting either directly on lymphocyte precursors or indirectly on a non–T cell subset that is critical for T‐cell maturation. In line with these findings in mice, Weekx et al [25] showed that, in the in vitro FTOC system, TNF promotes T cell differentiation of CD34 + cells from adult human BM, which we have also observed for human CB‐derived CD34 + cells (our unpublished data). Of interest, genetic polymorphism leading to increased TNFα production may enhance susceptibility to adult T‐cell leukemia/lymphoma among human T‐lymphotropic virus I carriers [34].…”

“…In mice, TNF induces CD25 (IL‐2R) expression on early immature thymocytes and is required for additional thymocyte maturation to single‐positive (SP) CD4 and CD8 cells [24]. Weekx et al [25] showed that TNF also promotes differentiation of human BM CD34 + ‐enriched cell populations into T cells in the FTOC system. However, T‐cell differentiation and selection, as well as the thymus architecture, are normal in TNF knockout mice.…”

Cord blood stem cell transplantation for haemoglobinopathies

“…Higher proliferative capacity of CFU-GM, BFU-E, CFU-GEMM; greater proportion of CD38 ) CD34 + cells* (Broxmeyer et al, , 1992Mayani et al, 1998;Nakahata & Ogawa, 1982) Superior engraftment of CB CD34 + cells in NOD/SCID mice* , (Larochelle et al, 1996;Noort et al, 2001;Wang et al, 1997) Superior migration of CB CD34 + cells* , (Voermans et al, 1999) Superior generation of B and T-lymphocyte progenitors* (Arakawa-Hoyt et al, 1999;Weekx et al, 2000) Immunological 'immaturity'* , : -lower numbers of CD4 + , CD8 + and CD3 + T-cells and higher CD4/CD8 ratio -higher proportion of naïve CD45RA + T-cells -lower Th-1 type cytokines, eg IFN-c and TNF-a -reduced capacity of dendritic cells to produce IL-12 -immature T cell gene expression profile -greater T cell receptor diversity (Bradley & Cairo, 2005;Chang et al, 2005;Goriely et al, 2001;Harris et al, 1992;Kaminski et al, 2003;Nomura et al, 2001;Rabian-Herzog et al, 1993;Risdon et al, 1995;Talvensaari et al, 2002) NK-cells with higher cytotoxic activity (Gardiner et al, 1998;Nomura et al, 2001;Rabian-Herzog et al, 1993) biological and clinical significance of these findings, particularly for regenerative medicine, is yet to be fully defined [reviewed in and (Riordan et al, 2007)] and is not further considered in this review.…”

Implication of Possible Therapies Targeted for the Tachykinergic System with the Biology of Neurokinin Receptors and Emerging Related Proteins

“…Common lymphoid progenitors mature towards B-, T-, natural killer and dendritic cells [6,7]. T-cell development involves a complex process beginning from common lymphoid progenitor to mature pro-T-cells, followed by maturation and selection mainly in the thymus [8][9][10].…”