Intestinal microfold cells (M cells) are an enigmatic lineage of intestinal epithelial cells that initiate mucosal immune responses through the uptake and transcytosis of luminal antigens. The mechanisms of M-cell differentiation are poorly understood, as the rarity of these cells has hampered analysis. Exogenous administration of the cytokine RANKL can synchronously activate M-cell differentiation in mice. Here we show the Ets transcription factor Spi-B was induced early during M-cell differentiation. Absence of Spi-B silenced the expression of various M-cell markers and prevented the differentiation of M cells in mice. The activation of T cells via an oral route was substantially impaired in the intestine of Spi-B-deficient (Spib−/−) mice. Our study demonstrates that commitment to the intestinal M-cell lineage requires Spi-B as a candidate master regulator.

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We thank Z. Guo, Y. Obata, Y. Oohara, Y. Fujimura, M. Ohmae, C. Uetake, Y. Usami and S. Kimura for technical support; T. Kawai for electron microscopy analysis; and H. Matsui (Kitasato University) for S. Typhimurium. Supported by RIKEN (T.Kan., D.T. and I.S.), the Kishimoto Foundation (K.Ho., I.S., H.H. and T.Kai.), the Ministry of Education, Culture, Sports, Science and Technology of Japan (21022049 to K.Ha.; 20060033, 21022048 and 21117003 to T.Kai.; and 20113003 to H.O.), the Japan Society for the Promotion of Science (22689017 to K.Ha.; 23790550 to T.Kan.; 21390155 to H.O.; and 20390146, 23390124 and 18590483 to T.Kai.), the Japan Science and Technology Agency (K.Ha.), The Japan Science Society (K.Ha.), The Takeda Science Foundation (H.O.), The Mitsubishi Foundation (H.O.), The Uehara Memorial Foundation (T.Kai.), the US National Institutes of Health (DK64730 to I.R.W.) and the Bill & Melinda Gates Foundation (OPP1006977 to I.R.W.).

Author information


  1. Laboratory for Epithelial Immunobiology, Research Center for Allergy and Immunology (RCAI), The Institute of Physical and Chemical Research (RIKEN), Kanagawa, Japan.

    • Takashi Kanaya
    • , Koji Hase
    • , Daisuke Takahashi
    • , Shinji Fukuda
    • , Kumiko Nakai
    • , Ayako Sakamoto
    • , Yuuki Kitahara
    • , Toshi Jinnohara
    •  & Hiroshi Ohno
  2. Graduate School of Nanobioscience, Yokohama City University, Kanagawa, Japan.

    • Takashi Kanaya
    • , Koji Hase
    • , Shinji Fukuda
    • , Toshi Jinnohara
    •  & Hiroshi Ohno
  3. Laboratory for Host Defense, Research Center for Allergy and Immunology, RIKEN, Kanagawa, Japan.

    • Katsuaki Hoshino
    • , Izumi Sasaki
    • , Hiroaki Hemmi
    •  & Tsuneyasu Kaisho
  4. Laboratory for Immune Regulation, World Premier International Research Center Initiative (WPI) Immunology Frontier Research Center, Osaka University, Osaka, Japan.

    • Katsuaki Hoshino
    • , Izumi Sasaki
    • , Hiroaki Hemmi
    •  & Tsuneyasu Kaisho
  5. Department of Allergy and Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan.

    • Izumi Sasaki
    •  & Tsuneyasu Kaisho
  6. Department of Pathology, Emory University School of Medicine, Atlanta, Georgia, USA.

    • Kathryn A Knoop
    • , Nachiket Kumar
    •  & Ifor R Williams
  7. Plant Science Center, RIKEN, Kanagawa, Japan.

    • Mayuko Sato
    •  & Kiminori Toyooka
  8. Department of Medicine and Clinical Oncology (K1), Graduate School of Medicine, Chiba University, Chiba, Japan.

    • Tatsuro Katsuno
    •  & Osamu Yokosuka
  9. Department of Immune Regulation, Graduate School of Medicine, Chiba University, Chiba, Japan.

    • Yuuki Kitahara
    •  & Hiroshi Ohno
  10. Center for Comparative Medicine, Department of Anatomy, Physiology & Cell Biology, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.

    • Stephen J McSorley


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I.R.W., K.Ha. and H.O. conceived of the study; T.Kan. designed and did the experiments, analyzed data and wrote the manuscript; K.Ha. contributed to adoptive-transfer experiments and data analysis; D.T. and Y.K. helped with flow cytometry; S.F. and T.J. helped with experiments involving infection with S. Typhimurium; K.N. and A.S. did expression analyses; K.Ho., I.S., H.H. and T.Kai. generated Spib−/− mice; K.A.K., N.K. and I.R.W. developed the protocol for treatment with RANKL; M.S. and K.T. helped with electron microscopy; O.Y. and T.Kat. provided human PP samples; S.J.M. provided SM1 mice; K.Ha. and I.R.W. edited the manuscript; and H.O. directed the research and edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Tsuneyasu Kaisho or Ifor R Williams or Hiroshi Ohno.

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