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J Exp Med. 1993 July 1; 178(1): 265–277.
PMCID: PMC2191096

Differentiation of CD3-4-8- human fetal thymocytes in vivo: characterization of a CD3-4+8- intermediate

Abstract

Human thymocyte differentiation was examined by injecting fetal thymic progenitor populations into human thymic xenografts in SCID-hu mice. Thymic progenitors were fluorescently labeled with the lipophilic dye PKH2. The phenotypes of their progeny could be identified by flow cytometric analysis of cells with a very high fluorescent PKH2 signal. Intrathymic injection of purified triple negative (TN) CD3-4-8- thymocytes resulted in the sequential appearance of CD3-4+8-, CD3-4+8+, and CD3+4+8+ cells, with the subsequent appearance of small numbers of phenotypically mature CD3+4+8- and CD3+4-8+ cells over a 4-d period. Sorted CD3-4+8- thymocytes injected intrathymically rapidly differentiated to CD4+8+ cells. CD4+8+ fetal thymocytes in cell cycle differentiated into phenotypically mature CD3+4+8- and CD3+4-8+ populations, whereas nondividing CD4+8+ cells failed to differentiate after intrathymic transfer. The number of cell divisions that occurred between the injection of TN thymocytes and their progeny at different time points was estimated based on the decrease in the intensity of the PKH2 label. The average length of the cell cycle for the TN population was calculated to be 24 h. The SCID-hu model thus provides a useful tool for studying the kinetics of cell division and differentiation of human thymocytes in vivo.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adkins B, Mueller C, Okada CY, Reichert RA, Weissman IL, Spangrude GJ. Early events in T-cell maturation. Annu Rev Immunol. 1987;5:325–365. [PubMed]
  • Spangrude GJ, Muller-Sieburg CE, Heimfeld S, Weissman IL. Two rare populations of mouse Thy-1lo bone marrow cells repopulate the thymus. J Exp Med. 1988 May 1;167(5):1671–1683. [PMC free article] [PubMed]
  • Owen JJ, Jenkinson EJ. Early events in T lymphocyte genesis in the fetal thymus. Am J Anat. 1984 Jul;170(3):301–310. [PubMed]
  • Baum CM, Weissman IL, Tsukamoto AS, Buckle AM, Peault B. Isolation of a candidate human hematopoietic stem-cell population. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2804–2808. [PubMed]
  • Guidos CJ, Weissman IL, Adkins B. Developmental potential of CD4-8- thymocytes. Peripheral progeny include mature CD4-8- T cells bearing alpha beta T cell receptor. J Immunol. 1989 Jun 1;142(11):3773–3780. [PubMed]
  • Guidos CJ, Weissman IL, Adkins B. Intrathymic maturation of murine T lymphocytes from CD8+ precursors. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7542–7546. [PubMed]
  • Guidos CJ, Danska JS, Fathman CG, Weissman IL. T cell receptor-mediated negative selection of autoreactive T lymphocyte precursors occurs after commitment to the CD4 or CD8 lineages. J Exp Med. 1990 Sep 1;172(3):835–845. [PMC free article] [PubMed]
  • Kappler JW, Roehm N, Marrack P. T cell tolerance by clonal elimination in the thymus. Cell. 1987 Apr 24;49(2):273–280. [PubMed]
  • Swat W, Dessing M, Baron A, Kisielow P, von Boehmer H. Phenotypic changes accompanying positive selection of CD4+CD8+ thymocytes. Eur J Immunol. 1992 Sep;22(9):2367–2372. [PubMed]
  • Fowlkes BJ, Schwartz RH, Pardoll DM. Deletion of self-reactive thymocytes occurs at a CD4+8+ precursor stage. Nature. 1988 Aug 18;334(6183):620–623. [PubMed]
  • Borgulya P, Kishi H, Müller U, Kirberg J, von Boehmer H. Development of the CD4 and CD8 lineage of T cells: instruction versus selection. EMBO J. 1991 Apr;10(4):913–918. [PubMed]
  • Weissman IL. Thymus cell migration. J Exp Med. 1967 Aug 1;126(2):291–304. [PMC free article] [PubMed]
  • Scollay RG, Butcher EC, Weissman IL. Thymus cell migration. Quantitative aspects of cellular traffic from the thymus to the periphery in mice. Eur J Immunol. 1980 Mar;10(3):210–218. [PubMed]
  • Egerton M, Shortman K, Scollay R. The kinetics of immature murine thymocyte development in vivo. Int Immunol. 1990;2(6):501–507. [PubMed]
  • Egerton M, Scollay R, Shortman K. Kinetics of mature T-cell development in the thymus. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2579–2582. [PubMed]
  • Baron C, Pénit C. Study of the thymocyte cell cycle by bivariate analysis of incorporated bromodeoxyuridine and DNA content. Eur J Immunol. 1990 Jun;20(6):1231–1236. [PubMed]
  • Spangrude GJ, Scollay R. Differentiation of hematopoietic stem cells in irradiated mouse thymic lobes. Kinetics and phenotype of progeny. J Immunol. 1990 Dec 1;145(11):3661–3668. [PubMed]
  • Terstappen LW, Huang S, Picker LJ. Flow cytometric assessment of human T-cell differentiation in thymus and bone marrow. Blood. 1992 Feb 1;79(3):666–677. [PubMed]
  • Swerdlow SH, Angermeier PA, Hartman AL. Intrathymic ontogeny of the T cell receptor associated CD3 (T3) antigen. Lab Invest. 1988 Apr;58(4):421–427. [PubMed]
  • Hori T, Cupp J, Wrighton N, Lee F, Spits H. Identification of a novel human thymocyte subset with a phenotype of CD3- CD4+ CD8 alpha + beta-1. Possible progeny of the CD3- CD4- CD8- subset. J Immunol. 1991 Jun 15;146(12):4078–4084. [PubMed]
  • Merkenschlager M, Fisher AG. Human postnatal thymocytes generate phenotypically immature CD3dim, CD5dim, CD1abright progeny in organ culture. J Immunol. 1992 Feb 15;148(4):1012–1015. [PubMed]
  • Denning SM, Jones DM, Ware RE, Weinhold KJ, Brenner MB, Haynes BF. Analysis of clones derived from human CD7+CD4-CD8-CD3- thymocytes. Int Immunol. 1991 Oct;3(10):1015–1024. [PubMed]
  • Denning SM, Kurtzberg J, Leslie DS, Haynes BF. Human postnatal CD4- CD8- CD3- thymic T cell precursors differentiate in vitro into T cell receptor delta-bearing cells. J Immunol. 1989 May 1;142(9):2988–2997. [PubMed]
  • Hori T, Spits H. Clonal analysis of human CD4-CD8-CD3- thymocytes highly purified from postnatal thymus. J Immunol. 1991 Apr 1;146(7):2116–2121. [PubMed]
  • Rouse RV, van Ewijk W, Jones PP, Weissman IL. Expression of MHC antigens by mouse thymic dendritic cells. J Immunol. 1979 Jun;122(6):2508–2515. [PubMed]
  • van Ewijk W. T-cell differentiation is influenced by thymic microenvironments. Annu Rev Immunol. 1991;9:591–615. [PubMed]
  • Haynes BF. Human thymic epithelium and T cell development: current issues and future directions. Thymus. 1990 Nov-Dec;16(3-4):143–157. [PubMed]
  • de la Hera A, Marston W, Aranda C, Toribio ML, Martinez C. Thymic stroma is required for the development of human T cell lineages in vitro. Int Immunol. 1989;1(5):471–478. [PubMed]
  • Merkenschlager M, Fisher AG. Selective manipulation of the human T-cell receptor repertoire expressed by thymocytes in organ culture. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4255–4259. [PubMed]
  • Fisher AG, Larsson L, Goff LK, Restall DE, Happerfield L, Merkenschlager M. Human thymocyte development in mouse organ cultures. Int Immunol. 1990;2(6):571–578. [PubMed]
  • McCune JM, Namikawa R, Kaneshima H, Shultz LD, Lieberman M, Weissman IL. The SCID-hu mouse: murine model for the analysis of human hematolymphoid differentiation and function. Science. 1988 Sep 23;241(4873):1632–1639. [PubMed]
  • McCune JM, Kaneshima H, Lieberman M, Weissman IL, Namikawa R. The scid-hu mouse: current status and potential applications. Curr Top Microbiol Immunol. 1989;152:183–193. [PubMed]
  • Waller EK, Sen-Majumdar A, Kamel OW, Hansteen GA, Schick MR, Weissman IL. Human T-cell development in SCID-hu mice: staphylococcal enterotoxins induce specific clonal deletions, proliferation, and anergy. Blood. 1992 Dec 15;80(12):3144–3156. [PubMed]
  • Waller EK, Kamel OW, Cleary ML, Majumdar AS, Schick MR, Lieberman M, Weissman IL. Growth of primary T-cell non-Hodgkin's lymphomata in SCID-hu mice: requirement for a human lymphoid microenvironment. Blood. 1991 Nov 15;78(10):2650–2665. [PubMed]
  • Battye FL, Shortman K. Flow cytometry and cell-separation procedures. Curr Opin Immunol. 1991 Apr;3(2):238–241. [PubMed]
  • Hollander Z, Loken MR. Simultaneous analysis of DNA content and surface antigens in human bone marrow. Cytometry. 1988 Sep;9(5):485–490. [PubMed]
  • Melnicoff MJ, Morahan PS, Jensen BD, Breslin EW, Horan PK. In vivo labeling of resident peritoneal macrophages. J Leukoc Biol. 1988 May;43(5):387–397. [PubMed]
  • Civin CI, Strauss LC, Brovall C, Fackler MJ, Schwartz JF, Shaper JH. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984 Jul;133(1):157–165. [PubMed]
  • Namikawa R, Weilbaecher KN, Kaneshima H, Yee EJ, McCune JM. Long-term human hematopoiesis in the SCID-hu mouse. J Exp Med. 1990 Oct 1;172(4):1055–1063. [PMC free article] [PubMed]
  • Katsura Y, Kina T, Amagai T, Tsubata T, Hirayoshi K, Takaoki Y, Sado T, Nishikawa SI. Limiting dilution analysis of the stem cells for T cell lineage. J Immunol. 1986 Oct 15;137(8):2434–2439. [PubMed]
  • Arase H, Fukushi N, Hatakeyama S, Ogasawara K, Iwabuchi K, Iwabuchi C, Negishi I, Good RA, Onoé K. Sequential analysis of the thymocyte differentiation in fully allogeneic bone marrow chimera in mice. II. Further characterization of the CD4+ or CD8+ single positive thymocytes. Immunobiology. 1990 Feb;180(2-3):167–183. [PubMed]
  • Vandekerckhove BA, Namikawa R, Bacchetta R, Roncarolo MG. Human hematopoietic cells and thymic epithelial cells induce tolerance via different mechanisms in the SCID-hu mouse thymus. J Exp Med. 1992 Apr 1;175(4):1033–1043. [PMC free article] [PubMed]
  • Wineman JP, Gilmore GL, Gritzmacher C, Torbett BE, Müller-Sieburg CE. CD4 is expressed on murine pluripotent hematopoietic stem cells. Blood. 1992 Oct 1;80(7):1717–1724. [PubMed]
  • Sen-Majumdar A, Lieberman M, Alpert S, Wiessman IL, Small M. Differentiation of CD3-4-8- thymocytes in short-term thymic stromal cell culture. J Exp Med. 1992 Aug 1;176(2):543–551. [PMC free article] [PubMed]
  • Fujiwara H, Ogata M, Mizushima Y, Tatsumi Y, Takai Y, Hamaoka T. Proliferation and differentiation of immature thymocytes induced by a thymic stromal cell clone. Thymus. 1990 Nov-Dec;16(3-4):159–172. [PubMed]
  • Hugo P, Waanders GA, Scollay R, Shortman K, Boyd RL. Ontogeny of a novel CD4+CD8-CD3- thymocyte subpopulation: a comparison with CD4- CD8+ CD3- thymocytes. Int Immunol. 1990;2(3):209–218. [PubMed]
  • Matsumoto K, Yoshikai Y, Matsuzaki G, Asano T, Nomoto K. A novel CD3-J11d+ subset of CD4+CD8- cells repopulating thymus in radiation bone marrow chimeras. Eur J Immunol. 1989 Jul;19(7):1203–1207. [PubMed]
  • Matsumoto K, Yoshikai Y, Moroi Y, Asano T, Ando T, Nomoto K. Two differential pathways from double-negative to double-positive thymocytes. Immunology. 1991 Jan;72(1):20–26. [PubMed]
  • Namikawa R, Kaneshima H, Lieberman M, Weissman IL, McCune JM. Infection of the SCID-hu mouse by HIV-1. Science. 1988 Dec 23;242(4886):1684–1686. [PubMed]
  • Richie ER, McEntire BB, Coghlan L, Poenie M. Murine T-lymphomas corresponding to the immature CD4-8+ thymocyte subset. Dev Immunol. 1991;1(4):255–263. [PMC free article] [PubMed]
  • London NJ, Thirdborough SM, Swift SM, Bell PR, James RF. The diabetic "human reconstituted" severe combined immunodeficient (SCID-hu) mouse: a model for isogeneic, allogeneic, and xenogeneic human islet transplantation. Transplant Proc. 1991 Feb;23(1 Pt 1):749–749. [PubMed]

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