Abstract
Introduction:
Umbilical cord blood contains relatively abundant primitive CD34+ hematopoietic progenitor cells which can differentiate into T lymphocytes ex vivo
Materials and Methods:
In this study, thymic stromal cells (TSCs) were isolated from aborted fetuses and a monolayer culture system was established. Highly-purified CD34+ cells from umbilical cord blood were cultured on the TSCs after limiting-dilution. The cells were then harvested and evaluated for CD4, CD8, and CD3 expression at different time points. CD4+CD8−CD3− lymphoid progenitor cells that could differentiate into mature T lymphocytes were observed after 15 days when a cocktail of cytokines, including Flt-3 ligand, stem cell factor, interleukin (IL)-12, and IL-2, was added.
Results:
These results thus show that CD4+CD8−CD3− cells can be derived from CD34+ cells in vitro when cultured on TSCs.
Conclusions:
We showed that CD4+CD8−CD3− cells can be derived from highly purified CD34+ cells on TSCs during T-cell lymphopoiesis in vitro.
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References
Andersnon G, Owen JJ, Moore NC et al (1994) Thymic epithelial cells provide unique signals for positive selection of CD4+CD8+ thymocytes in vitro. J Exp Med 179: 2027–2031
Brugnoni D, Airò P, Tosoni C et al (1997) CD3–CD4+ cells with a Th2-like pattern of cytokine production in the peripheral blood of a patient with cutaneous T cell lymphoma. Leukemia 11: 1983–1985
Deftos ML, Huang E, Ojala EW et al (2000) Notch1 signaling promotes the maturation of CD4 and CD8 SP thymocytes. Immunity 13: 73–84
Egerton M, Scollay R, Shortman K (1990) Kinetics of mature T-cell development in the thymus. Proc Natl Acad Sci USA 87: 2579–2582
Estes DM, Turaga PS, Sievers KM et al (1993) Characterization of an unusual cell type (CD4+CD3−) expanded by helminth infection and related to the parasite stress response. J Immunol 150: 1846–1856
Galy A, Verma S, Barcena A et al (1993) Precursors of CD3+CD4+CD8+ cells in the human thymus are defined by expression of CD34. Delineation of early events in human thymic development. J Exp Med 178: 391–401
Gardner JP, Rosenzweig M, Marks DF (1998) T-lymphopoietic capacity of cord blood-derived CD34+ progenitor cells. Exp Hematol 26: 991–999
Guidos CJ, Danska JS, Fathman CG et al (1990) T cell receptor- mediated negative selection of autoreactive T lymphocyte precursors occurs after commitment to the CD4 or CD8 lineages. J Exp Med 172: 835–845
Jenkinson EJ, Franchi LL, Kingstion R et al (1982) Effect of deoxyguanosine on lymphopoiesis in the developing thymus rudiment in vitro: Application in the production of chimeric thymus rudiments. Eur J Immunol 12: 583–587
Kakinuma S, Tanaka Y, Chinzei R et al (2003) Human umbilical cord blood as a source of transplantable hepatic progenitor cells. Stem Cells 21: 217–227
Kappler JW, Roehm N, Marrack P (1987) T cell tolerance by clonal elimination in the thymus. Cell 49: 273–280
Kelly KA, Scollay R (1992) Seeding of neonatal lymph nodes by T cells and identification of a novel population of CD3−CD4+ cells. Eur J Immunol 22: 329–334
Kim MY, Bekiaris V, McConnell FM et al (2005) OX40 signals during priming on dendritic cells inhibit CD4 T cell proliferation: IL-4 switches off OX40 signals enabling rapid proliferation of Th2 effectors. J Immunol 174: 1433–1437
Kim MY, McConnell FM, Gaspal FM et al (2007) Function of CD4+CD3− cells in relation to B- and T-zone stroma in spleen. Blood 109: 1602–1610
Kim MY, Toellner KM, White A et al (2006) Neonatal and adult CD4+CD3− cells share similar gene expression profile, and neonatal cells up-regulate OX40 ligand in response to TL1A (TNFSF15). J Immunol 177: 3074–3081
Kingston R, Jenkinson EJ, Owen JJ (1985) A single stem cell can recolonize an embryonic thymus, producing phenotypically distinct T-cell populations. Nature 317: 811–813
Kraft DL, Weissman IL, Waller EK (1993) Differentiation of CD3-4-8- human fetal thymocytes in vivo: characterization of a CD3-4+8– intermediate. J Exp Med 178: 265–277
La Motte-Mohs RN, Herer E, Zúñiga-Pflücker JC (2005) Induction of T-cell development from human cord blood hematopoietic stem cells by Delta-like 1 in vitro. Blood 105: 1431–1439
Lane PJ, Gaspal FM, Kim MY (2005) Two sides of a cellular coin: CD4 +CD3− cells regulate memory responses and lymphnode organization. Nat Rev Immunol 5: 655–660
Mikkola HK, Klintman J, Yang H et al (2003) Haematopoietic stem cells retain long-term repopulating activity and multipotency in the absence of stem-cell leukaemia SCL/tal-1 gene. Nature 421: 547–551
Mourcin F, Grenier N, Mayol JF et al (2005) Mesenchymal stem cells support expansion of in vitro irradiated CD34+ cells in the presence of SCF, FLT3 ligand, TPO and IL-3: potential application to autologous cell therapy in accidentally irradiated victims. Radiat Res 164: 1–9
Nanki T, Kohsaka H, Miyasaka N (1998) Development of human peripheral TCRBJ gene repertoire. J Immunol 161: 228–233
Nasu K, Matsubayashi H, Maeda Y et al (1994) A patient with adult T cell leukemia in smoldering stage expressing an aberrant phenotype of CD3− and CD4+. Rinsho Ketsueki 35: 774–779
Nelson BH, Willerford DM (1998) Biology of the interleukin-2 receptor. Adv Immunol 70: 1–81
Poznansky MC, Evans RH, Foxall RB et al (2000) Efficent generation of human cells from a tissue-engineered thymic organoid. Nat Biotechnol 18: 729–734
Rosenzweig M, Bunting EM, Damico RL et al (1994) Human neonatal thymic organ culture: an ex vivo model of thymocyte ontogeny and HIV-1 infection. Pathobiology 62: 245–251
Rosenzweig M, Yamada K, Harper DM et al (2001) T-cell differentiation of human and non-human primate CD34+hematopoietic progenitor cells using porcine thymic stroma. Xenotransplantation 8: 185–192
Sarun S, Dalloul AH, Laurent C et al (1998) Human CD34+ thymocyte maturation: pre-T and NK cell differentiation no neonatal thymic stromal cell culture. Cell Immunol 190: 23–32
Su DM, Manley NR (2002) Stage-specific changes in fetal thymocyte proliferation during the CD4−8− to CD4+8+ transition in wild type, Rag1–/–, and Hoxa3,Pax1 mutant mic. BMC Immunol 3: 12
Sugimoto K, Tamayose K, Sasaki M et al (2002) More than 13 years of hypereosinophila associated with clonal CD3−CD4+ lymphocytosis of TH2/TH0 type. Int J Hematol 75: 281–284
Swat W, Dessing M, Baron A et al (1992) Phenotypic changes accompanying positive selection of CD4+CD8+ thymocytes. Eur J Immunol 22: 2367–2372
Tse W, Laughlin MJ (2005) Umbilical cord blood transplantation: a new alternative option. Hematology Am Soc Hematol Educ Program 2005: 377–383
Utsuyama M, Shiraishi J, Takahashi H et al (2003) Glia maturation factor produced by thymic epithelial cells plays a role in T cell differentiation in the thymic microenvironment. Int Immunol 15: 557–564
Yeoman H, Gress RE, Bare CV et al (1993) Human bone marrow and umbilical cord blood cells generate CD4+ and CD8+ single-positive T cells in murine fetal thymus organ culture. Proc Nat Acad Sci USA 90: 10778–10782
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Jin, J.G., Bai, B.J., Yao, Z.J. et al. Detection of a CD4+CD8−CD3− cell subpopulation during the differentiation of cord blood CD34+ cells into T cells in vitro . Arch. Immunol. Ther. Exp. 57, 213–219 (2009). https://doi.org/10.1007/s00005-009-0022-0
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DOI: https://doi.org/10.1007/s00005-009-0022-0