In vitro co-culture systems for studying molecular basis of cellular interaction between Aire-expressing medullary thymic epithelial cells and fresh thymocytes

Yoshitaka Yamaguchi, Jun Kudo, Tetsuhiko Yoshida, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We previously established three mouse cell lines (Aire+ TEC1, Aire+ TEC2 and Aire+ DC) from the medullary thymic epithelial cells (mTECs) and dendritic cells (mDCs). These cells constitutively expressed ''autoimmune regulator (Aire) gene'' and they exhibited various features of self antigen-presenting cells (self-APCs) present in the thymic medullary region. Here, we confirmed our previous observation that Aire+ thymic epithelial cells adhere to fresh thymocytes and kill them by inducing apoptosis, thus potentially reproducing in vitro some aspects of the negative selection of T cells in vivo. In this system, a single Aire+ cell appeared able to kill ,30 thymocytes within 24 hrs. Moreover, we observed that ectopic expression of peripheral tissue-specific antigens (TSAs), and expression of several surface markers involved in mTEC development, increased as Aire+ cell density increases toward confluency. Thus, these Aire+ cells appear to behave like differentiating mTECs as if they pass through the developmental stages from intermediate state toward mature state. Surprisingly, an in vitro co-culture system consisting of Aire+ cells and fractionated sub-populations of fresh thymocytes implied the possible existence of two distinct subtypes of thymocytes (named as CD4+ killer and CD4- rescuer) that may determine the fate (dead or alive) of the differentiating Aire+ mTECs. Thus, our in vitro coculture system appears to mimic a part of ''in vivo thymic crosstalk''.

Original languageEnglish
Pages (from-to)1071-1082
Number of pages12
JournalBiology Open
Volume3
Issue number11
DOIs
Publication statusPublished - 2014 Nov 15

Fingerprint

thymocytes
coculture
Thymocytes
Coculture Techniques
epithelial cells
Epithelial Cells
cells
T-cells
antigen-presenting cells
Autoantigens
Antigen-Presenting Cells
dendritic cells
Regulator Genes
Crosstalk
regulator genes
Dendritic Cells
apoptosis
T-lymphocytes
Cell Count
Genes

Keywords

  • AIRE
  • Aire cell lines
  • Autoimmunity
  • Central selftolerance
  • Thymic crosstalk

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

In vitro co-culture systems for studying molecular basis of cellular interaction between Aire-expressing medullary thymic epithelial cells and fresh thymocytes. / Yamaguchi, Yoshitaka; Kudo, Jun; Yoshida, Tetsuhiko; Shimizu, Nobuyoshi.

In: Biology Open, Vol. 3, No. 11, 15.11.2014, p. 1071-1082.

Research output: Contribution to journalArticle

Yamaguchi, Yoshitaka ; Kudo, Jun ; Yoshida, Tetsuhiko ; Shimizu, Nobuyoshi. / In vitro co-culture systems for studying molecular basis of cellular interaction between Aire-expressing medullary thymic epithelial cells and fresh thymocytes. In: Biology Open. 2014 ; Vol. 3, No. 11. pp. 1071-1082.
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