Abstract
Flow cytometric and immunocytochemical analyses of murine fetal thymus (FT) cells with antibodies to various surface markers and transcription factors reveal that the synthesis of TCF-1 and GATA-3 protein begins simultaneously in a fraction of the most immature population of FT cells, which have the phenotype of CD4-CD8-CD44+CD25-. No TCF-1- producing cells is found in the fetal liver (FL). In CD44+CD25- FT cells, the production of TCF-1 is immediately followed by intracellular expression of CD3 epsilon. It is also found that the T cell development from FL, but not FT, progenitors in the FT organ culture system is severely inhibited by the addition of antisense oligonucleotides for either TCF-1 or GATA-3. These results strongly suggest that TCF-1 and GATA-3 play essential roles in the initiation of the earliest steps of T cell development in the thymus.
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- Amagai T., Itoi M., Kondo Y. Limited development capacity of the earliest embryonic murine thymus. Eur J Immunol. 1995 Mar;25(3):757–762. doi: 10.1002/eji.1830250320. [DOI] [PubMed] [Google Scholar]
- Amagai T., Kina T., Hirokawa K., Nishikawa S., Imanishi J., Katsura Y. Dysfunction of irradiated thymus for the development of helper T cells. J Immunol. 1987 Jul 15;139(2):358–364. [PubMed] [Google Scholar]
- Anderson G., Jenkinson E. J., Moore N. C., Owen J. J. MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus. Nature. 1993 Mar 4;362(6415):70–73. doi: 10.1038/362070a0. [DOI] [PubMed] [Google Scholar]
- Boyd R. L., Hugo P. Towards an integrated view of thymopoiesis. Immunol Today. 1991 Feb;12(2):71–79. doi: 10.1016/0167-5699(91)90161-L. [DOI] [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Dou Y. M., Watanabe Y., Aiba Y., Wada K., Katsura Y. A novel culture system for induction of T cell development: modification of fetal thymus organ culture. Thymus. 1994;23(3-4):195–207. [PubMed] [Google Scholar]
- Ferrick D. A., Schrenzel M. D., Mulvania T., Hsieh B., Ferlin W. G., Lepper H. Differential production of interferon-gamma and interleukin-4 in response to Th1- and Th2-stimulating pathogens by gamma delta T cells in vivo. Nature. 1995 Jan 19;373(6511):255–257. doi: 10.1038/373255a0. [DOI] [PubMed] [Google Scholar]
- Fontaine-Perus J. C., Calman F. M., Kaplan C., Le Douarin N. M. Seeding of the 10-day mouse embryo thymic rudiment by lymphocyte precursors in vitro. J Immunol. 1981 Jun;126(6):2310–2316. [PubMed] [Google Scholar]
- Fowlkes B. J., Edison L., Mathieson B. J., Chused T. M. Early T lymphocytes. Differentiation in vivo of adult intrathymic precursor cells. J Exp Med. 1985 Sep 1;162(3):802–822. doi: 10.1084/jem.162.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Georgopoulos K., Moore D. D., Derfler B. Ikaros, an early lymphoid-specific transcription factor and a putative mediator for T cell commitment. Science. 1992 Oct 30;258(5083):808–812. doi: 10.1126/science.1439790. [DOI] [PubMed] [Google Scholar]
- Godfrey D. I., Zlotnik A. Control points in early T-cell development. Immunol Today. 1993 Nov;14(11):547–553. doi: 10.1016/0167-5699(93)90186-O. [DOI] [PubMed] [Google Scholar]
- Gubbay J., Collignon J., Koopman P., Capel B., Economou A., Münsterberg A., Vivian N., Goodfellow P., Lovell-Badge R. A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature. 1990 Jul 19;346(6281):245–250. doi: 10.1038/346245a0. [DOI] [PubMed] [Google Scholar]
- Hashimoto Y., Blank K. J. T cell receptor genes and T cell development in virus-transformed early T cell lines. J Immunol. 1990 Feb 15;144(4):1518–1525. [PubMed] [Google Scholar]
- Ikuta K., Uchida N., Friedman J., Weissman I. L. Lymphocyte development from stem cells. Annu Rev Immunol. 1992;10:759–783. doi: 10.1146/annurev.iy.10.040192.003551. [DOI] [PubMed] [Google Scholar]
- Katsura Y., Kina T., Takaoki Y., Nishikawa S. Quantification of the progenitors for thymic T cells in various organs. Eur J Immunol. 1988 Jun;18(6):889–895. doi: 10.1002/eji.1830180609. [DOI] [PubMed] [Google Scholar]
- Keller G. Clonal analysis of hematopoietic stem cell development in vivo. Curr Top Microbiol Immunol. 1992;177:41–57. doi: 10.1007/978-3-642-76912-2_4. [DOI] [PubMed] [Google Scholar]
- Ko L. J., Yamamoto M., Leonard M. W., George K. M., Ting P., Engel J. D. Murine and human T-lymphocyte GATA-3 factors mediate transcription through a cis-regulatory element within the human T-cell receptor delta gene enhancer. Mol Cell Biol. 1991 May;11(5):2778–2784. doi: 10.1128/mcb.11.5.2778. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Landry D. B., Engel J. D., Sen R. Functional GATA-3 binding sites within murine CD8 alpha upstream regulatory sequences. J Exp Med. 1993 Sep 1;178(3):941–949. doi: 10.1084/jem.178.3.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lemischka I. R. What we have learned from retroviral marking of hematopoietic stem cells. Curr Top Microbiol Immunol. 1992;177:59–71. doi: 10.1007/978-3-642-76912-2_5. [DOI] [PubMed] [Google Scholar]
- Levelt C. N., Carsetti R., Eichmann K. Regulation of thymocyte development through CD3. II. Expression of T cell receptor beta CD3 epsilon and maturation to the CD4+8+ stage are highly correlated in individual thymocytes. J Exp Med. 1993 Dec 1;178(6):1867–1875. doi: 10.1084/jem.178.6.1867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsuzaki Y., Gyotoku J., Ogawa M., Nishikawa S., Katsura Y., Gachelin G., Nakauchi H. Characterization of c-kit positive intrathymic stem cells that are restricted to lymphoid differentiation. J Exp Med. 1993 Oct 1;178(4):1283–1292. doi: 10.1084/jem.178.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Molnár A., Georgopoulos K. The Ikaros gene encodes a family of functionally diverse zinc finger DNA-binding proteins. Mol Cell Biol. 1994 Dec;14(12):8292–8303. doi: 10.1128/mcb.14.12.8292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nikolić-Zugić J. Phenotypic and functional stages in the intrathymic development of alpha beta T cells. Immunol Today. 1991 Feb;12(2):65–70. doi: 10.1016/0167-5699(91)90160-u. [DOI] [PubMed] [Google Scholar]
- Ogawa M., Matsuzaki Y., Nishikawa S., Hayashi S., Kunisada T., Sudo T., Kina T., Nakauchi H., Nishikawa S. Expression and function of c-kit in hemopoietic progenitor cells. J Exp Med. 1991 Jul 1;174(1):63–71. doi: 10.1084/jem.174.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oosterwegel M., van de Wetering M., Dooijes D., Klomp L., Winoto A., Georgopoulos K., Meijlink F., Clevers H. Cloning of murine TCF-1, a T cell-specific transcription factor interacting with functional motifs in the CD3-epsilon and T cell receptor alpha enhancers. J Exp Med. 1991 May 1;173(5):1133–1142. doi: 10.1084/jem.173.5.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oosterwegel M., van de Wetering M., Timmerman J., Kruisbeek A., Destree O., Meijlink F., Clevers H. Differential expression of the HMG box factors TCF-1 and LEF-1 during murine embryogenesis. Development. 1993 Jun;118(2):439–448. doi: 10.1242/dev.118.2.439. [DOI] [PubMed] [Google Scholar]
- Owen J. J., Jenkinson E. J. Early events in T lymphocyte genesis in the fetal thymus. Am J Anat. 1984 Jul;170(3):301–310. doi: 10.1002/aja.1001700306. [DOI] [PubMed] [Google Scholar]
- Owen J. J., Jenkinson E. J. Embryology of the lymphoid system. Prog Allergy. 1981;29:1–34. [PubMed] [Google Scholar]
- Pandolfi P. P., Roth M. E., Karis A., Leonard M. W., Dzierzak E., Grosveld F. G., Engel J. D., Lindenbaum M. H. Targeted disruption of the GATA3 gene causes severe abnormalities in the nervous system and in fetal liver haematopoiesis. Nat Genet. 1995 Sep;11(1):40–44. doi: 10.1038/ng0995-40. [DOI] [PubMed] [Google Scholar]
- Papiernik M., Lepault F., Pontoux C. Synergistic effect of colony-stimulating factors and IL-2 on prothymocyte proliferation linked to the maturation of macrophage/dendritic cells within L3T4-Lyt-2-Ia-Mac- cells. J Immunol. 1988 Mar 1;140(5):1431–1434. [PubMed] [Google Scholar]
- Peschon J. J., Morrissey P. J., Grabstein K. H., Ramsdell F. J., Maraskovsky E., Gliniak B. C., Park L. S., Ziegler S. F., Williams D. E., Ware C. B. Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice. J Exp Med. 1994 Nov 1;180(5):1955–1960. doi: 10.1084/jem.180.5.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Péault B., Khazaal I., Weissman I. L. In vitro development of B cells and macrophages from early mouse fetal thymocytes. Eur J Immunol. 1994 Mar;24(3):781–784. doi: 10.1002/eji.1830240345. [DOI] [PubMed] [Google Scholar]
- Robey E., Fowlkes B. J. Selective events in T cell development. Annu Rev Immunol. 1994;12:675–705. doi: 10.1146/annurev.iy.12.040194.003331. [DOI] [PubMed] [Google Scholar]
- Rodewald H. R., Kretzschmar K., Swat W., Takeda S. Intrathymically expressed c-kit ligand (stem cell factor) is a major factor driving expansion of very immature thymocytes in vivo. Immunity. 1995 Sep;3(3):313–319. doi: 10.1016/1074-7613(95)90116-7. [DOI] [PubMed] [Google Scholar]
- Rodewald H. R. Pathways from hematopoietic stem cells to thymocytes. Curr Opin Immunol. 1995 Apr;7(2):176–187. doi: 10.1016/0952-7915(95)80002-6. [DOI] [PubMed] [Google Scholar]
- Saint-Ruf C., Ungewiss K., Groettrup M., Bruno L., Fehling H. J., von Boehmer H. Analysis and expression of a cloned pre-T cell receptor gene. Science. 1994 Nov 18;266(5188):1208–1212. doi: 10.1126/science.7973703. [DOI] [PubMed] [Google Scholar]
- Schreiber E., Matthias P., Müller M. M., Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. doi: 10.1093/nar/17.15.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
- Takebe Y., Seiki M., Fujisawa J., Hoy P., Yokota K., Arai K., Yoshida M., Arai N. SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol Cell Biol. 1988 Jan;8(1):466–472. doi: 10.1128/mcb.8.1.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Travis A., Amsterdam A., Belanger C., Grosschedl R. LEF-1, a gene encoding a lymphoid-specific protein with an HMG domain, regulates T-cell receptor alpha enhancer function [corrected]. Genes Dev. 1991 May;5(5):880–894. doi: 10.1101/gad.5.5.880. [DOI] [PubMed] [Google Scholar]
- Verbeek S., Izon D., Hofhuis F., Robanus-Maandag E., te Riele H., van de Wetering M., Oosterwegel M., Wilson A., MacDonald H. R., Clevers H. An HMG-box-containing T-cell factor required for thymocyte differentiation. Nature. 1995 Mar 2;374(6517):70–74. doi: 10.1038/374070a0. [DOI] [PubMed] [Google Scholar]
- Wada K., Kina T., Kawamoto H., Kondo M., Katsura Y. Requirement of cell interactions through adhesion molecules in the early phase of T cell development. Cell Immunol. 1996 May 25;170(1):11–19. doi: 10.1006/cimm.1996.0128. [DOI] [PubMed] [Google Scholar]
- Wang B., Biron C., She J., Higgins K., Sunshine M. J., Lacy E., Lonberg N., Terhorst C. A block in both early T lymphocyte and natural killer cell development in transgenic mice with high-copy numbers of the human CD3E gene. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9402–9406. doi: 10.1073/pnas.91.20.9402. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watanabe Y., Gyotoku J., Katsura Y. Analysis of the development of T cells by transferring precursors into cultured fetal thymus with a microinjector. Thymus. 1989;13(1-2):57–71. [PubMed] [Google Scholar]
- Watanabe Y., Katsura Y. Development of T cell receptor alpha beta-bearing T cells in the submersion organ culture of murine fetal thymus at high oxygen concentration. Eur J Immunol. 1993 Jan;23(1):200–205. doi: 10.1002/eji.1830230131. [DOI] [PubMed] [Google Scholar]
- Waterman M. L., Fischer W. H., Jones K. A. A thymus-specific member of the HMG protein family regulates the human T cell receptor C alpha enhancer. Genes Dev. 1991 Apr;5(4):656–669. doi: 10.1101/gad.5.4.656. [DOI] [PubMed] [Google Scholar]
- Wilson A., MacDonald H. R. Expression of genes encoding the pre-TCR and CD3 complex during thymus development. Int Immunol. 1995 Oct;7(10):1659–1664. doi: 10.1093/intimm/7.10.1659. [DOI] [PubMed] [Google Scholar]
- Wittwer C. T., Fillmore G. C., Garling D. J. Minimizing the time required for DNA amplification by efficient heat transfer to small samples. Anal Biochem. 1990 May 1;186(2):328–331. doi: 10.1016/0003-2697(90)90090-v. [DOI] [PubMed] [Google Scholar]
- Wittwer C. T., Fillmore G. C., Hillyard D. R. Automated polymerase chain reaction in capillary tubes with hot air. Nucleic Acids Res. 1989 Jun 12;17(11):4353–4357. doi: 10.1093/nar/17.11.4353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu L., Antica M., Johnson G. R., Scollay R., Shortman K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J Exp Med. 1991 Dec 1;174(6):1617–1627. doi: 10.1084/jem.174.6.1617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wurster A. L., Siu G., Leiden J. M., Hedrick S. M. Elf-1 binds to a critical element in a second CD4 enhancer. Mol Cell Biol. 1994 Oct;14(10):6452–6463. doi: 10.1128/mcb.14.10.6452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamamoto M., Ko L. J., Leonard M. W., Beug H., Orkin S. H., Engel J. D. Activity and tissue-specific expression of the transcription factor NF-E1 multigene family. Genes Dev. 1990 Oct;4(10):1650–1662. doi: 10.1101/gad.4.10.1650. [DOI] [PubMed] [Google Scholar]
- Yang Z., Gu L., Romeo P. H., Bories D., Motohashi H., Yamamoto M., Engel J. D. Human GATA-3 trans-activation, DNA-binding, and nuclear localization activities are organized into distinct structural domains. Mol Cell Biol. 1994 Mar;14(3):2201–2212. doi: 10.1128/mcb.14.3.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zúiga-Pflücker J. C., Jiang D., Lenardo M. J. Requirement for TNF-alpha and IL-1 alpha in fetal thymocyte commitment and differentiation. Science. 1995 Jun 30;268(5219):1906–1909. doi: 10.1126/science.7541554. [DOI] [PubMed] [Google Scholar]
- de Vries P., Brasel K. A., McKenna H. J., Williams D. E., Watson J. D. Thymus reconstitution by c-kit-expressing hematopoietic stem cells purified from adult mouse bone marrow. J Exp Med. 1992 Dec 1;176(6):1503–1509. doi: 10.1084/jem.176.6.1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Ewijk W. T-cell differentiation is influenced by thymic microenvironments. Annu Rev Immunol. 1991;9:591–615. doi: 10.1146/annurev.iy.09.040191.003111. [DOI] [PubMed] [Google Scholar]
- van Genderen C., Okamura R. M., Fariñas I., Quo R. G., Parslow T. G., Bruhn L., Grosschedl R. Development of several organs that require inductive epithelial-mesenchymal interactions is impaired in LEF-1-deficient mice. Genes Dev. 1994 Nov 15;8(22):2691–2703. doi: 10.1101/gad.8.22.2691. [DOI] [PubMed] [Google Scholar]
- van de Wetering M., Oosterwegel M., van Norren K., Clevers H. Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes. EMBO J. 1993 Oct;12(10):3847–3854. doi: 10.1002/j.1460-2075.1993.tb06063.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- von Boehmer H. Developmental biology of T cells in T cell-receptor transgenic mice. Annu Rev Immunol. 1990;8:531–556. doi: 10.1146/annurev.iy.08.040190.002531. [DOI] [PubMed] [Google Scholar]