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

Singh N, Seki Y, Takami M, Baban B, Chandler PR, Khosravi D, Zheng X, Takezaki M, Lee JR, Mellor AL, Bollag WB, Iwashima M
Source: Nat Methods
Publication Date: (2006)
Issue: 3(8): 629-36
Research Area:
Immunotherapy / Hematology
Cells used in publication:
T cell, mouse - BALB/c
Species: mouse
Tissue Origin: blood
T cell, mouse - C57BL/6
Species: mouse
Tissue Origin: blood
T cell, mouse, stim
Species: mouse
Tissue Origin: blood
Nucleofector® I/II/2b
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.