Change And Significance Of Cytokines Of Th17 And Treg In BALF Of CS Exposure Induced Mice COPD Model After The Adoptive Immunotherapy Based On DCs Over Expressed SOCS1

Objective:By reinfusing dendritic cells(DCs)highly expressed suppressor of cytokine signaling protein 1(SOCS1)into mice and observing the changes of cytokines of T helper17 cells(Th17)and regulatory T cells(Treg)in bronchoalveolar lavage fluid(BALF)of mice with chronic obstructive pulmonary disease(COPD),further explore the pathogenesis of COPD,looking for therapeutic targets for COPD.Methods:Cultured DCs cells in vitro and identified the phenotype and morphology.We constructed lentivirus over-expressed suppressor of cytokine signaling 1(SOCS1)and transfected immature DCs(imDCs)for fifth days to obtain DCs that over expressed SOCS1,used Western blot to detect the expression of SOCS1 in DCs,and injected cells into mice by tail vein injection;42 C57BL/6 mice were randomly divided into 7 groups,each group 6,the injected group of NS,imDC 1×10~6,DC-SOCS1 1×10~6,and DC-SOCS12×10~6 for the first day of smoking,the injected group of NS,imDC 1×10~6,DC-SOCS11×10~6,and DC-SOCS1 2×10~6 for the seventh day of smoking.The COPD mice model was established by smoke fumigation,and seven groups of mice were killed after the last fourth weeks(28 days)after the last fumigation,then collected BALF in mice.The changes of cytokines about Th17 and Treg in BALF were detected by enzyme linked immunosorbent assay(ELISA).Results:1.Mice bone marrow derived DCs were cultured in vitro.The expression of CD80 and CD83 was highly expressed in DCs cells used flow cytometry after lipopolysaccharide(LPS)induction for the seventh days.Scanning electron microscopy showed that there were many folds and dendritic processes on DCs surface after seventh days of LPS induction.2.The expression of SOCS1 in the positive transfected group was higher than that negative transfected group and the blank group by Western bloting,and the difference was statistically significant.3.the content of interleukin 17(IL-17)and IL-23 in injected group of imDC 1×10~6,DC-SOCS1 1×10~6 and DC-SOCS1 2×10~6 for the first and seventh days decreased than that NS transfusion group,the difference was statistically significant(P<0.01);the content of IL-17 and IL-23 in injected group of DC-SOCS1 1×10~6 and DC-SOCS1 2×10~6for the first and seventh days decreased than that imDC 1×10~6 group,the difference was statistically significant(P<0.01);the content of IL-17 and IL-23 in injected group of DC-SOCS1 2×10~6 decreased than that DC-SOCS1 1×10~6 group,the difference was statistically significant(P<0.01);the content of IL-17 in DC-SOCS1 1×10~6 transfusion group for the seventh day more than that injected for the first day,but the difference was not statistically significant(P>0.05),the content of IL-23 increased,the difference was statistically significant(P<0.01);the content of IL-17 and IL-23 in DC-SOCS1 2×10~6 transfusion group for the seventh day increased than that injected for the first day,the difference was statistically significant(P<0.01).4.the content of IL-10 and TGF-?in injected group of imDC 1×10~6,DC-SOCS11×10~6and DC-SOCS1 2×10~6for the first and seventh days more than that NS transfusion group,the difference was statistically significant(P<0.01);the content of IL-10 and transforming growth factor-beta(TGF-?)in injected group of DC-SOCS1 1×10~6 and DC-SOCS1 2×10~6for the first and seventh days more than that imDC 1×10~6group,the difference was statistically significant(P<0.01);the content IL-10 and TGF-?in injected group of DC-SOCS1 2×10~6more than that DC-SOCS1 1×10~6group,the difference was statistically significant(P<0.01);the content of IL-10 and TGF-?in DC-SOCS1 1×10~6transfusion group for the seventh day decreased than that injected for the first day,the difference was statistically significant(P<0.05,P<0.01);the content of IL-10 and TGF-?in DC-SOCS1 2×10~6 transfusion group for the seventh day decreased than that injected for the first day,the difference was statistically significant(P<0.01).Conclusion:1.Transfused imDCs cells into mice could inhibit production of cytokines related about Th17.2.DCs transfection of SOCS1 inhibited the Th17 based immune response in COPD,and the effect was better than that of imDCs transfusion alone,which was related to the concentration and time.It suggested that appropriate concentration and early intervention might inhibit the Th17 based immune response in COPD earlier,and provide a new target for COPD treatment.