Effects Of Prednisolone Combined With IL-2 Short-Term Treatment In Vitro On Treg Cells

Backgrounds and ObjectivesMany studies have shown that regulatory T cells(Tregs)are critical for limiting the effector T cells(Teffs)and maintaining autoimmune tolerance.Imbalances and dysfunction of Tregs in vivo can lead to immune-mediated diseases and tissue damage.In addition,Tregs are involved in a variety of immunotherapeutic mechanisms to reduce pathogenic immune responses.However,the effects of currently used anti-inflammatory drugs and immunomodulators on the function and homeostasis of Tregs are not fully understood.A large number of studies on interleukin 2(IL-2)and interleukin 2 receptor(IL2R)have revealed that it regulates immune tolerance by affecting the function and homeostasis of Tregs.The preservation or enhancement of Tregs functionality has become a key component of modern immunotherapeutic strategies.Low-dose IL-2 administration in vivo has been shown to be safe and effective in the expansion of Tregs,and studies have shown that glucocorticoids can enhance the expansion of Tregs cells in autoimmune patients.However,the direct effects of these immunomodulatory drugs on Tregs are still unknown.In the complex context of considering the etiology,pathogenesis and treatment of clinical diseases,the response between various drugs and the effects of drugs on immune function will be It is a factor affecting the disease,and it suggests that the clinical application is more cautious in solving the problem of abnormal immune function and immune response recovery.In this experiment,peripheral blood from healthy human was pretreated with prednisolone or prednisolone combined with IL-2.The cell ratio of Tregs,Th1 and Th17 cells was compared,and glucocorticoids and IL-2 were observed against Tregs,Th1 and Th17.Analysis cellular effects and the possible mechanisms of which glucocorticoids and IL-2 up-regulate or inhibit Tregs cells.MethodsThe peripheral blood of healthy people was taken and the lymphocytes were treated.The experiment was divided into prednisolone treatment group(GC group),prednisolone + high dose IL-2 treatment group(GC+IL-2hi group),prednisone + low-dose IL-2 treatment group(GC+IL-2low group)and control group(control group),after 4 days of treatment,multi-color analysis was used for staining.Flow cytometry was used to detect and analyze cell frequency changes of Treg(CD3+CD4+Foxp3+),Th1(CD3+CD4+foxp3-IFNγ+IL17-)and Th17(CD3+CD4+foxp3-IL17+IFNγ-),and the changes of the cell frequency of each group after Treg cells were grouped.Results1.Treg cell frequency: We find Treg cell frequency obviously lower in the control group than that in the GC group(p<0.001),and Treg cell frequency greatly increased in the GC+IL-2hi group and the GC+IL-2low group(p value 0.001,0.0001,respectively);Compared with the GC group,the GC+IL-2hi group and the GC+IL-2low group were greatly elevated(p<0.001).2.Th1 cell frequency: Compared with control group,Th1 cell frequency of GC group,GC+IL-2hi group and GC+IL-2low group were hugely lower(p<0.001);the comparable results of other groups were not statistically significant.3.Th17 cell frequency: Compared with control group,Th17 cell frequency of GC group,GC+IL-2hi group and GC+IL-2low were all decreased(p<0.05);the comparable results of other groups were not statistically significant.4.Treg cells subgroups:(1)Active Tregs cells frequency: Compared with the control group,active Tregs cell frequency of GC group decreased(p<0.001),but it increased in GC+IL-2hi group(p<0.05),especially greatly increased in GC+IL-2low group(p<0.001).Compared with the GC group,active Tregs cell frequency of GC+IL-2hi group and the GC+IL-2low group were substantially increased(p<0.001);the frequency in GC+IL-2low group substantially higher than the IL-2hi group(p < 0.001).(2)Resting Tregs cells frequency: compared with control group,resting Tregs cell frequency of GC+IL-2hi group increased(p<0.05);compared with GC group,the frequency in GC+IL-2low group greatly decreased(p<0.001);the cell frequency of the IL-2low group was substantially lower than that of the IL-2hi group(p<0.001).(3)Frequency of none Tregs cells: Compared with control group,none Tregs cells frequency of GC group increased(p<0.05),GC+IL-2hi group decreased(p<0.05),GC+IL-2low group greatly decreased(p<0.001).Compared with the GC group,the frequency of GC+IL-2hi group and the GC+IL-2low group were greatly decreased(p<0.001);the GC+IL-2hi group and the GC+IL-2low group were compared,the frequency of IL-2low group substantially lower than the IL-2hi group(p < 0.001).5.The average fluorescence intensity of CD25: Compared with the control group,the average fluorescence intensity of GC+IL-2hi group and the GC+IL-2low group were greatly increased(p<0.001);compared with the GC group,the average fluorescence intensity of GC+IL-2hi group and the GC+ IL-2low group was significantly elevated(p < 0.001).6.The average fluorescence intensity of CD39: Compared with the control group,the average fluorescence intensity of CD39 of GC group was greatly lower(p<0.001),and the average fluorescence intensity of CD39 of GC+IL-2hi group and the GC+IL-2low group were greatly higher(p<0.001);compared with the GC group,GC+IL-2hi group and GC+IL-2low group were greatly increased(p<0.001).Conclusions1,Short-term treatment of prednisolone greatly inhibits Treg cells,mainly inhibits the active Treg subgroup with immunomodulatory activity,and changes the balance of Th1/Th17 and Treg cells.2.IL-2 has a greatly proliferative effects on Treg cells and can reverse the Treg cell inhibitory effect caused by prednisolone.