Submitted on August 15, 2007
Accepted on June 9, 2008
Differentiation of a murine intestinal epithelial cell line (MIE) towards the M cell lineage
Takashi Kanaya1, Kohtaro Miyazawa2, Ikuro Takakura1, Wataru Itani1, Kouichi Watanabe1, Shyuichi Ohwada1, Haruki Kitazawa3, Michael T Rose4, Huw R McConochie4, Hideyuki Okano5, Takahiro Yamaguchi1, and Hisashi Aso1*
1 Graduate School of Agricultural Science, Tohoku University, Cellular Biology Laboratory, Sendai, Miyagi, Japan
2 Graduate School of Agricultural Science, Tohoku University, Cellular Biology Laboratory, Sendai, Miyagi, Japan; Department of Surgery, Yale Medical School, Section of Neuropathology, New Haven, Connecticut, United States
3 Graduate School of Agricultural Science, Tohoku University, Laboratory of Animal Products Chemistry, Sendai, Miyagi, Japan
4 University of Wales, Institute of Rural Sciences, United Kingdom
5 Keio University, School of Medicine, Department of Physiology, Tokyo, Japan
* To whom correspondence should be addressed. E-mail: asosan{at}bios.tohoku.ac.jp.
M cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer's patches (PP). These cells can transport antigens and microorganisms into underlying lymphoid tissues. Despite the important role of M cells in mucosal immune responses, the origin and mechanisms of differentiation as well as cell death of M cells remain unclear. In order to clarify the mechanism of M cell differentiation, we established a novel murine intestinal epithelial cell line (MIE) from the C57BL/6 mouse. MIE cells grow rapidly and have a cobblestone morphology, which is a typical feature of intestinal epithelial cells. Additionally, they express cytokeratin, villin, cell-cell junctional proteins, and alkaline phosphatase activity, and can form microvilli. Their expression of Musashi-1 antigen indicates that they may be close to intestinal stem cells or transit-amplifying cells. MIE cells are able to differentiate into the M cell lineage following co-culture with intestinal lymphocytes, but not with Peyer's patch lymphocytes (PPL). However, PPL co-stimulated with anti-CD3/CD28 mAbs caused MIE cells to display typical features of M cells, such as transcytosis activity, the disorganization of microvilli and the expression of M cell markers. This transcytosis activity of MIE cells was not induced by T cells isolated from PPL co-stimulated with the same mAbs, and was reduced by the depletion of the T cell population from PPL. A mixture of T cells treated with mAbs and B cells both from PPL led MIE cells to differentiate into M cells. We report here that MIE cells have the potential ability to differentiate into M cells, and that this differentiation required activated T cells and B cells.
