Enrichment of regulatory CD4+CD25+ T cells by inhibition of phospholipase D signaling

Nagendra Singh, Yoichi Seki, Mariko Takami, Babak Baban, Phil R. Chandler, Davood Khosravi, Xiangjian Zheng, Mayuko Takezaki, Jeffrey R. Lee, Andrew L. Mellor, Wendy B. Bollag, Makio Iwashima

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Antigen stimulation of lymphocytes induces upregulation of phospholipase D (PLD) activity, but the biological significance of PLD-mediated signaling in T cells has not been well established. Here we demonstrate that PLD signaling is essential for proliferation of mouse CD8+ T cells and CD4+CD25- T cells, but is not required for proliferation of CD4+ CD25+ regulatory T cells. We exploited this observation to develop an efficient method to enrich for regulatory T cells starting from preparations of total CD4+ T lymphocytes. Inhibition of PLD signaling blocked effector T-cell proliferation after T cell-antigen receptor (TCR) engagement, but had no significant effect on the proliferation of CD4+CD25+ T cells with regulatory functions. Consequently, cells expanded in vitro for one week by antigen receptor stimulation with PLD signal inhibition were markedly enriched for regulatory T cells.

Original languageEnglish (US)
Pages (from-to)629-636
Number of pages8
JournalNature Methods
Volume3
Issue number8
DOIs
StatePublished - Aug 1 2006

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Singh, N., Seki, Y., Takami, M., Baban, B., Chandler, P. R., Khosravi, D., Zheng, X., Takezaki, M., Lee, J. R., Mellor, A. L., Bollag, W. B., & Iwashima, M. (2006). Enrichment of regulatory CD4+CD25+ T cells by inhibition of phospholipase D signaling. Nature Methods, 3(8), 629-636. https://doi.org/10.1038/nmeth903