Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Deletion of the CD4 silencer element supports a stochastic mechanism of thymocyte lineage commitment

Abstract

The mechanism of T cell lineage commitment remains controversial; to examine it we deleted the CD4-silencer element in the germ line of a mouse using a combination of gene targeting and Cre/LoxP-mediated recombination. We found that these mice were unable to extinguish CD4 expression either in immature thymocytes or mature CD8+ cytotoxic T cells (CTLs), which resulted in the development of major histocompatibility complex class II–restricted double-positive CTLs in the periphery. This finding strongly supports a stochastic over an instructive mechanism of coreceptor down-regulation.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Gene-targeting at the CD4 locus.
Figure 2: Expression of CD4 on lymph node T cells.
Figure 3: Mature DP thymocytes in CD4Δsil mice.
Figure 4: Mature DP T cells in CD4Δsil mice.
Figure 5: MHC class II–restriction of DP cells.
Figure 6: Mature DP thymocytes and lymph node T cells in CD4Δsil mice that lacked MHC class I.

References

  1. Jorgensen, J. L., Reay, P. A., Ehrich, E. W. & Davis, M. M. Molecular components of T-cell recognition. Annu. Rev. Immunol. 10, 835–873 (1992).

    Article  CAS  PubMed  Google Scholar 

  2. Robey, E. & Fowlkes, B. J. Selective events in T cell development. Annu. Rev. Immunol. 12, 675–705 (1994).

    Article  CAS  PubMed  Google Scholar 

  3. Marrack, P. & Kappler, J. Positive selection of thymocytes bearing αβ T cell receptors. Curr. Opin. Immunol. 9, 250–255 (1997).

    Article  CAS  PubMed  Google Scholar 

  4. von Boehmer, H. & Kisielow, P. Lymphocyte lineage commitment: instruction versus selection. Cell 73, 207–208 (1993).

    Article  Google Scholar 

  5. Davis, C. B. & Littman, D. R. Thymocyte lineage commitment: is it instructed or stochastic? Curr. Opin. Immunol. 6, 266–272 (1994).

    Article  CAS  PubMed  Google Scholar 

  6. Chan, S. H., Benoist, C. & Mathis, D. A challenge to the instructive model of positive selection. Immunol. Rev. 135, 119–131 (1993).

    Article  CAS  PubMed  Google Scholar 

  7. Chan, S. H., Cosgrove, D., Waltzinger, C., Benoist, C. & Mathis, D. Another view of the selective model of thymocyte selection. Cell 73, 225–236 (1993).

    Article  CAS  PubMed  Google Scholar 

  8. van Meerwijk, J. P. & Germain, R. N. Development of mature CD8+ thymocytes: selection rather than instruction? Science 261, 911–915 (1993).

    Article  CAS  PubMed  Google Scholar 

  9. Itano, A., Kioussis, D. & Robey, E. Stochastic component to development of class I major histocompatibility complex-specific T cells. Proc. Natl Acad. Sci. USA 91, 220–224 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Robey, E., Itano, A., Fanslow, W. C. & Fowlkes, B. J. Constitutive CD8 expression allows inefficient maturation of CD4+ helper T cells in class II major histocompatibility complex mutant mice. J. Exp. Med. 179, 1997–2004 (1994).

    Article  CAS  PubMed  Google Scholar 

  11. Baron, A., Hafen, K. & von Boehmer, H. A human CD4 transgene rescues CD4CD8+ cells in β 2-microglobulin-deficient mice. Eur. J. Immunol. 24, 1933–1936 (1994).

    Article  CAS  PubMed  Google Scholar 

  12. Corbella, P. et al. Functional commitment to helper T cell lineage precedes positive selection and is independent of T cell receptor MHC specificity. Immunity 1, 269–276 (1994).

    Article  CAS  PubMed  Google Scholar 

  13. Paterson, R. K. et al. Thymic development in human CD4 transgenic mice. Positive selection occurs after commitment to the CD8 lineage. J. Immunol. 153, 3491–3503 (1994).

    CAS  PubMed  Google Scholar 

  14. von Boehmer, H. The selection of the αβ heterodimeric T cell receptor for antigen. Immunol. Today 7, 333–336 (1986).

    Article  CAS  Google Scholar 

  15. Teh, H. S. et al. Thymic major histocompatibility complex antigens and the αβ T-cell receptor determine the CD4/CD8 phenotype of T cells. Nature 335, 229–233 (1988).

    Article  CAS  PubMed  Google Scholar 

  16. Robey, E. A. et al. Thymic selection in CD8 transgenic mice supports an instructive model for commitment to a CD4 or CD8 lineage. Cell 64, 99–107 (1991).

    Article  CAS  PubMed  Google Scholar 

  17. Borgulya, P., Kishi, H., Muller, U., Kirberg, J. & von, B. H. Development of the CD4 and CD8 lineage of T cells: instruction versus selection. EMBO J. 10, 913–918 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Davis, C. B., Killeen, N., Crooks, M. E., Raulet, D. & Littman, D. R. Evidence for a stochastic mechanism in the differentiation of mature subsets of T lymphocytes. Cell 73, 237–247 (1993).

    Article  CAS  PubMed  Google Scholar 

  19. Chan, S. H., Waltzinger, C., Baron, A., Benoist, C. & Mathis, D. Role of coreceptors in positive selection and lineage commitment. EMBO J. 13, 4482–4489 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Crump, A. L., Grusby, M. J., Glimcher, L. H. & Cantor, H. Thymocyte development in major histocompatibility complex-deficient mice: evidence for stochastic commitment to the CD4 and CD8 lineages. Proc. Natl Acad. Sci. USA 90, 10739–10743 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kydd, R., Lundberg, K., Vremec, D., Harris, A. W. & Shortman, K. Intermediate steps in thymic positive selection. Generation of CD48+ T cells in culture from CD4+8+, CD4int8+, and CD4+8int thymocytes with up-regulated levels of TCR- CD3. J. Immunol. 155, 3806–3814 (1995).

    CAS  PubMed  Google Scholar 

  22. Lundberg, K., Heath, W., Kontgen, F., Carbone, F. R. & Shortman, K. Intermediate steps in positive selection: differentiation of CD4+8int TCRint thymocytes into CD48+TCRhi thymocytes. J. Exp. Med. 181, 1643–1651 (1995).

    Article  CAS  PubMed  Google Scholar 

  23. Suzuki, H., Punt, J. A., Granger, L. G. & Singer, A. Asymmetric signaling requirements for thymocyte commitment to the CD4+ versus CD8+ T cell lineages: a new perspective on thymic commitment and selection. Immunity 2, 413–425 (1995).

    Article  CAS  PubMed  Google Scholar 

  24. Lucas, B. & Germain, R. N. Unexpectedly complex regulation of CD4/CD8 coreceptor expression supports a revised model for CD4+CD8+ thymocyte differentiation. Immunity 5, 461–477 (1996).

    Article  CAS  PubMed  Google Scholar 

  25. Benveniste, P., Knowles, G. & Cohen, A. CD8/CD4 lineage commitment occurs by an instructional/default process followed by positive selection. Eur. J. Immunol. 26, 461–471 (1996).

    Article  CAS  PubMed  Google Scholar 

  26. Yasutomo, K., Doyle, C., Miele, L. & Germain, R. N. The duration of antigen receptor signaling determines CD4+ versus CD8+ T-cell lineage fate. Nature 404, 506–510 (2000).

    Article  CAS  PubMed  Google Scholar 

  27. Hernandez-Hoyos, G., Sohn, S. J., Rothenberg, E. V. & Alberola-Ila, J. Lck activity controls CD4/CD8 T cell lineage commitment. Immunity 12, 313–322 (2000).

    Article  CAS  PubMed  Google Scholar 

  28. Legname, G. et al. Inducible expression of a p56Lck transgene reveals a central role for Lck in the differentiation of CD4 SP thymocytes. Immunity 12, 537–546 (2000).

    Article  CAS  PubMed  Google Scholar 

  29. Brugnera, E. et al. Coreceptor reversal in the thymus: signaled CD4+8+ thymocytes initially terminate CD8 transcription even when differentiating into CD8+ T cells. Immunity 13, 59–71 (2000).

    Article  CAS  PubMed  Google Scholar 

  30. Sternberg, N. & Hamilton, D. Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites. J. Mol. Biol. 150, 467–486 (1981).

    Article  CAS  PubMed  Google Scholar 

  31. Sawada, S., Scarborough, J. D., Killeen, N. & Littman, D. R. A lineage-specific transcriptional silencer regulates CD4 gene expression during T lymphocyte development. Cell 77, 917–929 (1994).

    Article  CAS  PubMed  Google Scholar 

  32. Siu, G., Wurster, A. L., Duncan, D. D., Soliman, T. M. & Hedrick, S. M. A transcriptional silencer controls the developmental expression of the CD4 gene. EMBO J. 13, 3570–3579 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. McCready, P. M., Hansen, R. K., Burke, S. L. & Sands, J. F. Multiple negative and positive cis-acting elements control the expression of the murine CD4 gene. Biochim. Biophys. Acta 1351, 181–191 (1997)

    Article  CAS  PubMed  Google Scholar 

  34. Rushton, J. J., Zorich, G. P., Stolc, V. and Neudorf, S. M. Characterization of a promoter within the first intron of the human CD4 gene. Eur. J. Biochem. 245, 768–773 (1997)

    Article  CAS  PubMed  Google Scholar 

  35. Liao, N. S., Maltzman, J. & Raulet, D. H. Positive selection determines T cell receptor Vβ14 gene usage by CD8+ T cells. J. Exp. Med. 170, 135–143 (1989).

    Article  CAS  PubMed  Google Scholar 

  36. Jameson, S. C., Kaye, J. & Gascoigne, N. R. A T cell receptor Vα region selectively expressed in CD4+ cells. J. Immunol. 145, 1324–1331 (1990).

    CAS  PubMed  Google Scholar 

  37. Oxenius, A. et al. Presentation of endogenous viral proteins in association with major histocompatibility complex class II: on the role of intracellular compartmentalization, invariant chain and the TAP transporter system. Eur. J. Immunol. 25, 3402–3411 (1995)

    Article  CAS  PubMed  Google Scholar 

  38. Bosselut, R., Feigenbaum, L., Sharrow, S. O. and Singer, A. Strength of signaling by CD4 and CD8 coreceptor tails determines the number but not the lineage direction of positively selected thymocytes. Immunity 14, 483–494 (2001).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank H. Waldmann, R. Lechler and R. Zinkernagel for advice and helpful discussions; J. Rossant for the D3 embryonic stem cell line; K. Rajewski for the Cre-Lox cassettes; and R. Sumner and C. Hetherington for help with some blastocyst injections. Supported by the Croucher Foundation (R. L) and the Wellcome Trust (K. T and A. R).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Amin Rahemtulla.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leung, R., Thomson, K., Gallimore, A. et al. Deletion of the CD4 silencer element supports a stochastic mechanism of thymocyte lineage commitment. Nat Immunol 2, 1167–1173 (2001). https://doi.org/10.1038/ni733

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni733

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing