Ctla 4 ig

BALB/c mice were sensitized via the transfusion of allogeneic splenocytes from C57BL/6 mice that were separated by erythrocyte lysis (blood sample taken via the tail vein) on day-7. Cellular activity was found to be >99% via trypan blue staining and a total of 1×106 splenocytes were subsequently injected via the tail vein into BALB/c mice to establish the sensitized model. These mice were treated with IDO and iNOS inhibitors, respectively, on days −7 to 0 following intravenous injection with CTLA4Ig (Abcam, Cambridge, MA, USA) and anti-CD154 (500 µg/mouse, ab2391; eBioscience; Thermo Fisher Scientific, Inc., Waltham, MA, USA) on day −7.
The mice were divided into 4 groups. In the first group, each mouse was injected with 10 mg 1-methyl-DL-tryptophan (1-MT; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) i.p. The second group received 50 mg/kg AG (Sigma Aldrich; Merck KGaA) i.p., and the third group was treated with 1-MT+AG (as described above). The fourth group served as a negative control and these mice were injected with an equal volume of PBS. There were 10 mice in each group, and the injections were performed every other day (Fig. 1).