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Regulation of T cell receptor β allelic exclusion at a level beyond accessibility

Nature Immunology volume 6, pages 189–197 (2005)Cite this article

Abstract

Allelic exclusion of Vβ-to-DJβ recombination depends on asynchronous rearrangement of alleles of the gene encoding T cell receptor β in double-negative thymocytes and feedback inhibition that is maintained in double-positive thymocytes. Feedback is thought to be enforced through downregulation of Vβ accessibility. In an attempt to override this negative regulation, we introduced the enhancer of the gene encoding T cell receptor α into the Vβ gene cluster downstream of Vβ12. In double-negative thymocytes, the introduced enhancer had no measurable effect on accessibility, but Vβ12 rearrangement was stimulated and Vβ12 allelic exclusion was partially subverted. In contrast, double-positive thymocytes showed increased Vβ transcription and accessibility, but feedback inhibition of Vβ-to-DJβ recombination remained intact. Our results indicate additional regulatory constraints on Vβ-to-DJβ recombination that operate beyond the accessibility barrier.

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Figure 1: Tcrb locus and gene targeting.
Figure 2: Germline transcription in EαKI thymocytes.
Figure 3: Tcrb locus histone modifications in EαKI thymocytes.
Figure 4: Nucleosome positioning at the Vβ12 RSS in EαKI thymocytes.
Figure 5: Restriction endonuclease accessibility of Vβ gene segments in EαKI thymocytes.
Figure 6: Regulation of VDJβ rearrangement in EαKI thymocytes.
Figure 7: Vβ expression in EαKI mice.
Figure 8: Dual Vβ expression in EαKI mice.

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GenBank/EMBL/DDBJ

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Acknowledgements

We thank L. Martinek of the Duke University Cancer Center Flow Cytometry Facility for help with cell sorting and analysis; T. Wehrly and H. Boutrid for technical assistance; and Y. Zhuang, Y.-W. He and G. Kelsoe for critical review of the manuscript. Supported by the National Institutes of Health (AI49934 to M.S.K. and NIAID-T32 AI052077 to H.D.K.) and National Science Foundation (A.J.).

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Authors and Affiliations

  1. The Department of Immunology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Annette Jackson, Hrisavgi D Kondilis & Michael S Krangel

  2. Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, 63110, Missouri, USA

    Bernard Khor & Barry P Sleckman

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Correspondence to Michael S Krangel.

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Supplementary information

Supplementary Fig. 1

DNase Ihypersensitivity of Eαi in DN and DP thymocytes of EαKI mice. (PDF 119 kb)

Supplementary Fig. 2

Strategy for restriction endonuclease accessibility assay. (PDF 45 kb)

Supplementary Table 1

Oligonucleotides used as PCR primers or radiolabeled probes. (PDF 44 kb)

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Jackson, A., Kondilis, H., Khor, B. et al. Regulation of T cell receptor β allelic exclusion at a level beyond accessibility. Nat Immunol 6, 189–197 (2005). https://doi.org/10.1038/ni1157

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