Combination of RAPA plus IL-2 reduced production of IFN-γ and TNF-α. BALB/c mice (Thy1.2 and H-2k^d) were injected with 5 × 10⁶ TCD-BM cells from GFP transgenic C57BL/6 mice (Thy1.1 and H2-k^b), 5 × 10⁵ Tregs from wild-type C57BL/6 mice (Thy1.2 and H-2k^b) and 5 × 10⁵ CD25-depleted Tcons (CD4⁺CD8⁺CD25⁻ T cells) from congenic Thy1.1 C57BL/c mice (Thy1.1 and H2-k^b) after lethal irradiation with 800 cGy on day 0. Mice of each group received PBS, RAPA or IL-2 alone, or a combination of RAPA + IL-2 from day 0 to day 7 after BMT. Intracellular cytokines (A) were determined from splenocytes of each mice group, and serum was collected and cytokines (B) were analyzed in a multiplex assay system on day 7 after BMT. RAPA plus IL-2 resulted in the greatest reduction in the production of both IFN-γ and TNF-α assessed by intracellular cytokine production and reduction in the concentration of both of these cytokines in serum of animals. Results were shown as mean ± SD using horizontal column bar graphs (5 mice were included at each group). *P < .05, *** P < .001.