Enrichment of regulatory CD4(+)CD25(+) T cells by inhibition of phospholipase D signaling.
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.
Singh
Nagendra
N
Immunotherapy Center, Institute of Molecular Medicine and Genetics, Department of Medicine, Medical College of Georgia, 1120 15th Street, Augusta, Georgia 30912, USA.
Seki
Yoichi
Y
Takami
Mariko
M
Baban
Babak
B
Chandler
Phil R
PR
Khosravi
Davood
D
Zheng
Xiangjian
X
Takezaki
Mayuko
M
Lee
Jeffrey R
JR
Mellor
Andrew L
AL
Bollag
Wendy B
WB
Iwashima
Makio
M
D015704:CD4 Antigens / Q000276:immunology*; D018791:CD4 Lymphocyte Count; D002469:Cell Separation / Q000379:methods*; D010739:Phospholipase D / Q000276:immunology*; D015375:Receptors, Interleukin-2 / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology*
D015704:CD4 Antigens; D015375:Receptors, Interleukin-2; D010739:Phospholipase D