This observation underlines the need to be cautious to extrapolate in vitro studies with Tregs to in vivo situations. Besides the issue of level of FOXP3 expression, duration of expression may be an important facet determining the function of

induced Tregs. The reduced effectiveness of the induced FOXP3 T cells may be time-dependent as earlier in vitro studies report that continuous levels of FOXP3 are required to convert naive T cells into Tregs with full effectiveness 6. In this setting of systemic inflammation, 24 h seems to be too short to procure the full molecular and transcriptional changes necessary for suppression. On the other hand, it does seem to be sufficient to inhibit the cell from dividing after TCR stimulation in vitro. Accordingly, FOXP3 may act as an intrinsic regulator during inflammation, preventing collateral damage by temporarily silencing activated T DNA Methyltransferas inhibitor cells. In conclusion, during systemic inflammation due to cardiac surgery in children, FOXP3+ T cells lose suppressive capacity. While these cells are anergic to TCR stimulation, the transiently increased expressed FOXP3 is not capable of taking on a suppressive function. Furthermore, the inflammatory milieu in which Tregs exert their action after cardiac surgery inhibits

their suppressive activity. This study illustrates the functionality of FOXP3+ T cells in a human model of inflammation and underlines the requirement of more human in vivo systems to understand the properties and potential of induced FOXP3+ Tregs in human disease. Children admitted to our hospital 5-Fluoracil datasheet for surgical repair of either a VSD or an ASD were enrolled in this study. Patients were excluded from the study if at the time of admission they had received steroids within 2 wk before surgery, had signs of infection or had a documented immunodeficiency. Informed consent was obtained from the parents of children Thiamet G participating in the study. The medical ethics committee approved this study (METC 03/049-K, UMC Utrecht, The Netherlands). General anesthesia was always implemented using a standard technique

involving high-dose sufentanil, midazolam, pancuronium, dopamine and milrinone. All patients were given a single dose of dexamethason (1 mg/kg) after induction of anesthesia. Non-pulsatile CPB was used, the standard pump flow rate was 2.8 L/m2/min. Combined alpha and pH stat management of acid–base status was used during CPB. The cardioplegia procedure was standardized using St. Thomas’ solution. After weaning from CPB, all patients remained intubated and ventilated and were admitted to the pediatric intensive care for further management. All patients were treated by the same surgical team. Blood samples were obtained from a central venous catheter at the following time points: immediately after insertion during anesthetic induction (T1), at the end of the CPB (T2) and at 4 h (T3), 24 h (T4) and 48 h after surgery (T5).