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. 1990 Dec 1;172(6):1583–1588. doi: 10.1084/jem.172.6.1583

Lineage relationships and developmental kinetics of immature thymocytes: CD3, CD4, and CD8 acquisition in vivo and in vitro

PMCID: PMC2188740  PMID: 2147945

Abstract

T lymphocytes develop in the thymus from immunologically naive bone marrow precursors. Based on T cell receptor rearrangement and transcription, and thymic reconstitution potential, we have deduced a developmental sequence among immature thymocytes, before the acquisition of the lineage markers CD3, CD4, and CD8. In the current study, we have followed the ontogenic progression of the latter stages in this sequence, using two different systems: (a) in vivo, by direct injection into the thymus of nonirradiated, congenic recipients; and (b) in vitro, using culture medium without mitogens or cytokines. In vivo, the less mature Pgp-1- interleukin 2 receptor alpha-positive (IL- 2R alpha+) CD3-4-8- subset (also heat-stable antigen high) requires 3 d before becoming predominantly IL-2R alpha- CD3lo4+ 8+ typical cortical- type cells, and at least 5 d before the appearance of any mature single- positive cells (CD3hi4+ 8- or CD3hi4-8+). However, these Pgp-1- IL-2R alpha+ precursors do not differentiate further in unstimulated culture. The more mature Pgp-1- IL-2R alpha- CD3-4-8- subset becomes primarily CD3lo4+ 8+ within 1 d after transplantation, and some mature single- positive progeny are evident by day 3. By 5 d, most of these Pgp-1-IL- 2R alpha- precursor cells have become CD3hi, and have lost or are downregulating either CD4 or CD8. In culture, these Pgp-1- IL-2R alpha- cells also acquire high levels of CD4 and CD8 within 1 d, and low levels of CD3 by 2 d. However, they do not progress further to mature single positives in vitro, and most of them die by day 3. These experiments directly confirm our previously proposed developmental sequence, and demonstrate the kinetics of T lymphocyte production in a low-stress, steady-state environment.

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Selected References

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  1. Berg L. J., Pullen A. M., Fazekas de St Groth B., Mathis D., Benoist C., Davis M. M. Antigen/MHC-specific T cells are preferentially exported from the thymus in the presence of their MHC ligand. Cell. 1989 Sep 22;58(6):1035–1046. doi: 10.1016/0092-8674(89)90502-3. [DOI] [PubMed] [Google Scholar]
  2. Ceredig R., Dialynas D. P., Fitch F. W., MacDonald H. R. Precursors of T cell growth factor producing cells in the thymus: ontogeny, frequency, and quantitative recovery in a subpopulation of phenotypically mature thymocytes defined by monoclonal antibody GK-1.5. J Exp Med. 1983 Nov 1;158(5):1654–1671. doi: 10.1084/jem.158.5.1654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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. doi: 10.1073/pnas.87.7.2579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Guidos C. J., Danska J. S., Fathman C. G., Weissman I. L. 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. doi: 10.1084/jem.172.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guidos C. J., Weissman I. L., Adkins B. Intrathymic maturation of murine T lymphocytes from CD8+ precursors. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7542–7546. doi: 10.1073/pnas.86.19.7542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Howe R. C., MacDonald H. R. Heterogeneity of immature (Lyt-2-/L3T4-) thymocytes. Identification of four major phenotypically distinct subsets differing in cell cycle status and in vitro activation requirements. J Immunol. 1988 Feb 15;140(4):1047–1055. [PubMed] [Google Scholar]
  8. Hugo P., Waanders G. A., Scollay R., Shortman K., Boyd R. L. Ontogeny of a novel CD4+CD8-CD3- thymocyte subpopulation: a comparison with CD4- CD8+ CD3- thymocytes. Int Immunol. 1990;2(3):209–218. doi: 10.1093/intimm/2.3.209. [DOI] [PubMed] [Google Scholar]
  9. MacDonald H. R., Budd R. C., Howe R. C. A CD3- subset of CD4-8+ thymocytes: a rapidly cycling intermediate in the generation of CD4+8+ cells. Eur J Immunol. 1988 Apr;18(4):519–523. doi: 10.1002/eji.1830180405. [DOI] [PubMed] [Google Scholar]
  10. 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. doi: 10.1002/eji.1830190708. [DOI] [PubMed] [Google Scholar]
  11. Nikolic-Zugic J., Moore M. W. T cell receptor expression on immature thymocytes with in vivo and in vitro precursor potential. Eur J Immunol. 1989 Oct;19(10):1957–1960. doi: 10.1002/eji.1830191030. [DOI] [PubMed] [Google Scholar]
  12. Ohashi P. S., Pircher H., Bürki K., Zinkernagel R. M., Hengartner H. Distinct sequence of negative or positive selection implied by thymocyte T-cell receptor densities. Nature. 1990 Aug 30;346(6287):861–863. doi: 10.1038/346861a0. [DOI] [PubMed] [Google Scholar]
  13. Pearse M., Wu L., Egerton M., Wilson A., Shortman K., Scollay R. A murine early thymocyte developmental sequence is marked by transient expression of the interleukin 2 receptor. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1614–1618. doi: 10.1073/pnas.86.5.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Scollay R., Bartlett P., Shortman K. T cell development in the adult murine thymus: changes in the expression of the surface antigens Ly2, L3T4 and B2A2 during development from early precursor cells to emigrants. Immunol Rev. 1984 Dec;82:79–103. doi: 10.1111/j.1600-065x.1984.tb01118.x. [DOI] [PubMed] [Google Scholar]
  15. Scollay R., Wilson A., D'Amico A., Kelly K., Egerton M., Pearse M., Wu L., Shortman K. Developmental status and reconstitution potential of subpopulations of murine thymocytes. Immunol Rev. 1988 Aug;104:81–120. doi: 10.1111/j.1600-065x.1988.tb00760.x. [DOI] [PubMed] [Google Scholar]
  16. Scott B., Blüthmann H., Teh H. S., von Boehmer H. The generation of mature T cells requires interaction of the alpha beta T-cell receptor with major histocompatibility antigens. Nature. 1989 Apr 13;338(6216):591–593. doi: 10.1038/338591a0. [DOI] [PubMed] [Google Scholar]
  17. Shortman K., Wilson A., Egerton M., Pearse M., Scollay R. Immature CD4- CD8+ murine thymocytes. Cell Immunol. 1988 May;113(2):462–479. doi: 10.1016/0008-8749(88)90042-1. [DOI] [PubMed] [Google Scholar]
  18. Wilson A., D'Amico A., Ewing T., Scollay R., Shortman K. Subpopulations of early thymocytes. A cross-correlation flow cytometric analysis of adult mouse Ly-2-L3T4-(CD8-CD4-) thymocytes using eight different surface markers. J Immunol. 1988 Mar 1;140(5):1461–1469. [PubMed] [Google Scholar]
  19. Wilson A., Petrie H. T., Scollay R., Shortman K. The acquisition of CD4 and CD8 during the differentiation of early thymocytes in short-term culture. Int Immunol. 1989;1(6):605–612. doi: 10.1093/intimm/1.6.605. [DOI] [PubMed] [Google Scholar]

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