Figure 4.

Expression of transcription factor, cytokine, and cytokine receptor mRNA in Ts1 cells re-cultured with alloantigen and either rIL-2, rIL-12p70, or both rIL-2 and rIL-12p70. Ts1 cells re-cultured with alloantigen and either rIL-12p70 alone or rIL-2 and rIL-12p70 had high expression of foxp3 suggesting cells in all cultures were Treg. il-12rβ2 levels were preserved in cultures with rIL-2 alone and rIL-12p70 alone but not those cultured with both rIL-2 and rIL-12p70. ifngr expression was enhanced with rIL-2, and was sustained with rIL-12p70 but not with both rIL-2 and rIL-12p70. il-5 expression was increased with rIL-12p70 but waned with rIL-2 and was lost with rIL-2 and rIL-12p70. Induction of t-bet, the Th1 transcription factor, and ifn-γ occurred after re-culture with rIL-12p70 alone, and not after re-culture with rIL-2 or both rIL-2 and rIL-12p70. Thus, Ts1 cells re-cultured with rIL-12p70 alone developed a Th1-like Treg phenotype, while retaining foxp3 expression and the CD4⁺CD25⁺Foxp3⁺ phenotype. Re-culture of Ts1 cells with rIL-2 retained the Ts1 phenotype. Re-culture of Ts1 cells with both rIL-2 and rIL-12p70 led to loss of the Ts1 phenotype and did not induce a Th1-like Treg phenotype.