Gene expression profile of isolated splenic NK cells from mice treated with ptaquiloside and/or selenium. Mus musculus

To better understand the immunosuppressor mechanism of ptaquiloside in splenic NK cells and the reversion of this effect by selenium, we have employed whole genome microarray expression profile to identify genes associated with immunosuppression. Among 89 genes induced by ptaquiloside treatment in splenic NK cells only two genes (Mt1 and Mt2) were identified as related with its immunosuppressor effect. Moreover this augmented expression of Mt1 and Mt2 was totally abrogated by selenium co-treatment. These results were confirmed by flow cytometry in splenic cells harvested from other six mice and treated in vitro for 1 hour with ptaquiloside and/or selenium. Overall design: Twenty mice were separated randomly into four groups as Control (water), Pt (ptaquiloside 5.3 mg/kg), PtSe (ptaquiloside 5.3 mg/kg and selenium 1.3 mg/kg) and Se (selenium 1.3 mg/kg) and were treated daily by gavage for 14 days. After treatment, untouched NK cells were isolated using the NK cell isolation kit, LS columns, and QuadroMACS cell separator system (Miltenyi Biotec, Inc.) to perform RNA isolation and whole-genome gene expression profile. Thereby, five independent experiments were performed per group using different donors for each experiment. To confirm the increase of metallothionein protein induced by ptaquiloside, splenic cells were harvested from other six mice and treated in vitro for 1 hour with ptaquiloside [4.4 µg/mL] and/or selenium [0.1 mM] and analyzed by flow cytometry.