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. 1992 Sep 1;176(3):845–853. doi: 10.1084/jem.176.3.845

Studies on T cell maturation on defined thymic stromal cell populations in vitro

PMCID: PMC2119352  PMID: 1512547

Abstract

We describe an in vitro system in which positive selection of developing T cells takes place on defined stromal cell preparations, which include major histocompatibility complex class II+ epithelial cells but exclude cells of bone marrow origin. In this system, maturation of double-positive T cell receptor negative (TCR-), CD4+8+ thymocytes into single-positive TCR+, CD4+ and CD8+ cells takes place together with the development of functional competence. As in vivo, this maturation is associated with the upregulation of TCR levels as cells progress from double-positive to single-positive status. We also show that class II+ epithelial cells in these cultures are less efficient than dendritic cells in mediating the deletion (negative selection) of V beta 8+ cells by the superantigen staphylococcal enterotoxin B. For the first time, this approach provides a model in which the cellular interactions involved in both positive and negative selection can be studied under controlled in vitro conditions.

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Selected References

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  1. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  2. Benoist C., Mathis D. Positive selection of the T cell repertoire: where and when does it occur? Cell. 1989 Sep 22;58(6):1027–1033. doi: 10.1016/0092-8674(89)90501-1. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bill J., Palmer E. Positive selection of CD4+ T cells mediated by MHC class II-bearing stromal cell in the thymic cortex. Nature. 1989 Oct 19;341(6243):649–651. doi: 10.1038/341649a0. [DOI] [PubMed] [Google Scholar]
  5. Breel M., Mebius R. E., Kraal G. Dendritic cells of the mouse recognized by two monoclonal antibodies. Eur J Immunol. 1987 Nov;17(11):1555–1559. doi: 10.1002/eji.1830171105. [DOI] [PubMed] [Google Scholar]
  6. Carding S. R., Hayday A. C., Bottomly K. Cytokines in T-cell development. Immunol Today. 1991 Jul;12(7):239–245. doi: 10.1016/0167-5699(91)90037-T. [DOI] [PubMed] [Google Scholar]
  7. Ceredig R. Intrathymic proliferation of perinatal mouse alpha beta and gamma delta T cell receptor-expressing mature T cells. Int Immunol. 1990;2(9):859–867. doi: 10.1093/intimm/2.9.859. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Houssaint E., Flajnik M. The role of thymic epithelium in the acquisition of tolerance. Immunol Today. 1990 Oct;11(10):357–360. doi: 10.1016/0167-5699(90)90141-u. [DOI] [PubMed] [Google Scholar]
  10. Hugo P., Boyd R. L., Waanders G. A., Petrie H. T., Scollay R. Timing of deletion of autoreactive V beta 6+ cells and down-modulation of either CD4 or CD8 on phenotypically distinct CD4+8+ subsets of thymocytes expressing intermediate or high levels of T cell receptor. Int Immunol. 1991 Mar;3(3):265–272. doi: 10.1093/intimm/3.3.265. [DOI] [PubMed] [Google Scholar]
  11. Hünig T., Mitnacht R. T cell receptor-mediated selection of functional rat CD8 T cells from defined immature thymocyte precursors in short-term suspension culture. J Exp Med. 1991 Mar 1;173(3):561–568. doi: 10.1084/jem.173.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jenkinson E. J., Kingston R., Owen J. J. Newly generated thymocytes are not refractory to deletion when the alpha/beta component of the T cell receptor is engaged by the superantigen staphylococcal enterotoxin B. Eur J Immunol. 1990 Nov;20(11):2517–2520. doi: 10.1002/eji.1830201125. [DOI] [PubMed] [Google Scholar]
  13. Jenkinson E. J., Kingston R., Smith C. A., Williams G. T., Owen J. J. Antigen-induced apoptosis in developing T cells: a mechanism for negative selection of the T cell receptor repertoire. Eur J Immunol. 1989 Nov;19(11):2175–2177. doi: 10.1002/eji.1830191132. [DOI] [PubMed] [Google Scholar]
  14. Kingston R., Jenkinson E. J., Owen J. J. Characterization of stromal cell populations in the developing thymus of normal and nude mice. Eur J Immunol. 1984 Nov;14(11):1052–1056. doi: 10.1002/eji.1830141117. [DOI] [PubMed] [Google Scholar]
  15. Kisielow P., von Boehmer H. Negative and positive selection of immature thymocytes: timing and the role of the ligand for alpha beta T cell receptor. Semin Immunol. 1990 Jan;2(1):35–44. [PubMed] [Google Scholar]
  16. Larsson L., Timms E., Blight K., Restall D. E., Jat P. S., Fisher A. G. Characterization of murine thymic stromal-cell lines immortalized by temperature-sensitive simian virus 40 large T or adenovirus 5 E1a. Dev Immunol. 1991;1(4):279–293. doi: 10.1155/1991/14636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lorenz R. G., Allen P. M. Thymic cortical epithelial cells lack full capacity for antigen presentation. Nature. 1989 Aug 17;340(6234):557–559. doi: 10.1038/340557a0. [DOI] [PubMed] [Google Scholar]
  18. Mazda O., Watanabe Y., Gyotoku J., Katsura Y. Requirement of dendritic cells and B cells in the clonal deletion of Mls-reactive T cells in the thymus. J Exp Med. 1991 Mar 1;173(3):539–547. doi: 10.1084/jem.173.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Palacios R., Studer S., Samaridis J., Pelkonen J. Thymic epithelial cells induce in vitro differentiation of PRO-T lymphocyte clones into TCR alpha,beta/T3+ and TCR gamma,delta/T3+ cells. EMBO J. 1989 Dec 20;8(13):4053–4063. doi: 10.1002/j.1460-2075.1989.tb08589.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ramsdell F., Lantz T., Fowlkes B. J. A nondeletional mechanism of thymic self tolerance. Science. 1989 Nov 24;246(4933):1038–1041. doi: 10.1126/science.2511629. [DOI] [PubMed] [Google Scholar]
  21. Scollay R., Shortman K. Thymocyte subpopulations: an experimental review, including flow cytometric cross-correlations between the major murine thymocyte markers. Thymus. 1983 Sep;5(5-6):245–295. [PubMed] [Google Scholar]
  22. Tatsumi Y., Kumanogoh A., Saitoh M., Mizushima Y., Kimura K., Suzuki S., Yagi H., Horiuchi A., Ogata M., Hamaoka T. Differentiation of thymocytes from CD3-CD4-CD8- through CD3-CD4-CD8+ into more mature stages induced by a thymic stromal cell clone. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2750–2754. doi: 10.1073/pnas.87.7.2750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Van Vliet E., Jenkinson E. J., Kingston R., Owen J. J., Van Ewijk W. Stromal cell types in the developing thymus of the normal and nude mouse embryo. Eur J Immunol. 1985 Jul;15(7):675–681. doi: 10.1002/eji.1830150707. [DOI] [PubMed] [Google Scholar]
  24. White J., Herman A., Pullen A. M., Kubo R., Kappler J. W., Marrack P. The V beta-specific superantigen staphylococcal enterotoxin B: stimulation of mature T cells and clonal deletion in neonatal mice. Cell. 1989 Jan 13;56(1):27–35. doi: 10.1016/0092-8674(89)90980-x. [DOI] [PubMed] [Google Scholar]
  25. 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]

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