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Medullary thymic epithelial cells expressing Aire represent a unique lineage derived from cells expressing claudin

Nature Immunology volume 8, pages 304–311 (2007)Cite this article

Abstract

The autoimmune regulator Aire is expressed in a small proportion of medullary thymic epithelial cells (mTECs) and is crucial in the induction of central T cell tolerance. The origin and development of Aire+ mTECs, however, are not well understood. Here we demonstrate that the tight-junction components claudin-3 and claudin-4 (Cld3,4) were 'preferentially' expressed in Aire+ mTECs. In early ontogeny, Cld3,4hi TECs derived from the most apical layer of the stratified thymic anlage first expressed known mTEC markers such as UEA-1 ligand and MTS10. We provide evidence that such Cld3,4hi UEA-1+ TECs represented the initial progenitors specified for Aire+ mTECs, whose development crucially required NF-κB-inducing kinase and the adaptor molecule TRAF6. Our results suggest that Aire+ mTECs represent terminally differentiated cells in a unique lineage arising during thymic organogenesis.

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Figure 1: Expression of tight-junction components in a small population of mTECs.
Figure 2: Cld3,4+ TECs express Aire and large amounts of MHC class II and costimulatory molecules.
Figure 3: Expression of Cld3,4 marks the origin and development of mTECs during ontogeny.
Figure 4: C-CPEhi UEA-1+ TECs develop earlier but proliferate more slowly during embryonic ontogeny than do C-CPElo UEA-1+ TECs.
Figure 5: C-CPEhi UEA-1+ TECs sorted at E13.5 'preferentially' generate Cld3,4+ MTS10− mTECs expressing Aire.
Figure 6: Cld3,4+ Aire+ mTECs arise from C-CPEhi UEA-1+ TECs at E13.5 with cell proliferation.
Figure 7: Embryonic development of Cld3,4+ mTECs is profoundly impaired in aly/aly and TRAF6-deficient mice.

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Acknowledgements

We thank S. Tsukita and M. Furuse for providing claudin-transfected L cells; Y. Horiguchi (Research Institute for Microbial Diseases, Osaka University) for the C-CPE plasmid; H. Kawamoto and K. Masuda for discussions; and W.T.V. Germeraad for proofreading the manuscript. Supported by the Ministry of Education, Culture, Science, Sports and Technology of the Japanese government and Shimizu Foundation for Immunology Research.

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Authors and Affiliations

  1. Department of Immunology and Cell Biology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto, 606-8501, Japan

    Yoko Hamazaki & Nagahiro Minato

  2. Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8501, Japan

    Harumi Fujita & Nagahiro Minato

  3. Medical Institute of Bioregulation, Kyushu University, Higashi-ku, Fukuoka, 812-8582, Japan

    Takashi Kobayashi

  4. Department of Pathology and Lab Medicine, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Yongwon Choi

  5. The Walter and Eliza Hall Institute of Medical Research, 3050, Victoria, Australia

    Hamish S Scott

  6. Institute for Enzyme Research, University of Tokushima, Tokushima, 770-8503, Japan

    Mitsuru Matsumoto

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Contributions

Y.H. principally contributed to general experiments and design; H.F. contributed to flow cytometry; T.K. and Y.C. contributed to TRAF6-deficient mice; H.S. contributed to rat monoclonal antibodies to mouse Aire; M.M. contributed to Aire-deficient mice and discussions; and N.M. provided the overall design of the work.

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Correspondence to Nagahiro Minato.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Specific binding of recombinant C-CPE to claudin-3, 4 on the epithelial cell surface. (PDF 341 kb)

Supplementary Fig. 2

Rare Cld3, 4− Aire+ cells express CD11c. (PDF 533 kb)

Supplementary Fig. 3

Phenotypes of the sorted fractions from the B6 thymic stroma cells at E13.5. (PDF 560 kb)

Supplementary Fig. 4

Anti-H-2Kb antibody specifically detects both B6 donor-derived mTECs and cTECs in the reconstituted thymus. (PDF 991 kb)

Supplementary Fig. 5

Transfer of the Cld3,4low TECs at E13.5 results in the generation of all types of TECs including Cld3,4high Aire+ mTECs. (PDF 1473 kb)

Supplementary Table 1

PCR primers. (PDF 20 kb)

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Hamazaki, Y., Fujita, H., Kobayashi, T. et al. Medullary thymic epithelial cells expressing Aire represent a unique lineage derived from cells expressing claudin. Nat Immunol 8, 304–311 (2007). https://doi.org/10.1038/ni1438

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