Fig. 5

Inhibition of CDP-choline pathway by etomoxir prolonged the survival of the mAITL preclinical model. A Expression levels of CPT1a (inhibited by etomoxir) for the different T-cell populations (AITL patient Tfh cells (n = 8), healthy Tfh cells (n = 12), central memory T cells (Tcm) (n = 6), effector memory T cells (Tem) (n = 6), naive T cells (Tn) (n = 6), regulatory T cells (T reg) (n = 13) and stem memory T cell (Tscm) (n = 6) (mean ± SD, ***p < 0.001, ****p < 0.0001, ns: non-significant). B Splenic lymphoma cell from plck-GAPDH mice were injected intravenously into recipient NSG mice (n = 11), which were treated with vehicle (n = 5) of with the Cept1 inhibitor etomoxir (n = 6). Survival curves for both mice groups are shown in (C). Mice were sacrificed at humane endpoint or 140 days post transplant (*p < 0.05, Mantel-Cox test). D FACS analysis of percentage of CD4 + PD1.^high cells per total CD4 + T cells in the spleen of the indicated treatment groups at sacrifice; data are summarized in the histogram (mean ± SD, Vehicle. n = 5, Etomoxir n = 6), ***p < 0.001). E FACS analysis of percentage of GC B cells (GL-7 + CD95 +) on total B cells in the spleen of the indicated treatment groups at sacrifice; data are summarized in the histogram (mean ± SD, Vehicle n = 5, Etomoxir n = 6, ****p < 0.0001). F FACS analysis of CD4 + T cells stained for mitochondrial content by Mitotracker green (MTG) in the spleen of the indicated treatment groups at sacrifice; data are summarized in the histogram (mean ± SD, vehicle n = 5, Etomoxir n = 6, ****p < 0.0001). G FACS analysis of CD4 + T cells stained for ROS content by CellROX probe in the spleen of the indicated treatment groups at sacrifice; data are summarized in the histogram (mean ± SD, vehicle n = 5, etomoxir n = 6, ****p < 0.0001). H Splenocytes isolated from etomoxir or control-treated mAITL engrafted NSG mice were activated for 6 h with PMA/ionomycin in presence of golgi-stop, then surface stained for CD8 followed by intracellular staining for INFγ, perforin and granzyme B and analysed by FACS (mean ± SD, n = 4, *p < 0.05,**p < 0.01, ****p < 0.0001). (I) mAITL CD4 + T cells were treated with etomoxir at the indicated doses in the absence (-NAC) or presence of N-Acetyl-L-cysteine (+ NAC; 2 mM) for 96 h followed by FACS analysis for the CD4 + T-cell survival (mean ± SD, n = 3, ***p < 0.001, ****p < 0.0001, ns non significant)