A Study On The Participation Of Th17 Intervened By DCs That Overexpress SOCS1 In Immunoregulation Of COPD Mice

Objective:By culturing immature DCs(imDCs)that overexpressing SOCS1 in vitro,different types of DCs and different concentrations of DCs-SOCS1 were injected into COPD mice to observe the changes of Th17 cytokines in BALF and lung tissues.Methods:1.Bone marrow cells were washed from the tibia and femur of mice,and imDCs were induced by IL-4 and granulocyte-macrophage colony stimulating factors.2.The expression of SOCS1 on DCs-SOCS1 was detected by WB in the imDCs with overexpression of SOCS1,which were isolated and cultured on the fifth day without LPS stimulation3.On the seventh day of cell culture,LPS was added to induce maturation.The surface molecular markers CD80,CD83 and cell morphology were identified by flow cytometry and scanning electron microscopy respectively4.The 42 C57BL/6 mice were randomly divided into 7 groups,each group has 6,on day1 NS group,on day 1 imDCs(1 x 10~6)group,on day 1 smoking DCs-SOCS1(1 x 10~6)group,on day 7 DCs-SOCS1(1 x 10~6)group,on day 7 DCS-SOCS1(2 x 10~6)group,on day 7imDCs(1 x 10~6)group.The COPD mouse model was established by fumigation method.The mice were sacrificed within 24 hours of the last fumigation in 4 weeks(28 days)and BALF and lung tissue were collected.The changes of Th17-associated cytokines(IL-21)and IL-21/TGF-βin lung tissues and BALF were detected by ELISA.Results:1.Bone marrow-derived dendritic cells were successfully extracted,cultured and identified;2.SOCS1 lentivirus effectively transfected into DCs;3.The expression of SOCS1 in infected cells was significantly improved;4.imDCs or DCs-SOCS1 transitive immunity had an effect on the cytokine IL-21 secreted by Th17 and IL21/TGFβin the COPD mice BALF and lung tissues.(1)Compared with the control group,the IL-21 secretion decreased by injecting 1×10~6imDCs、1×10~6DCs-SOCS1(1×10~6)on the 1st and 7th day of fumigation(P<0.01);(2)Compared with control group,the concentration of IL-21 in BALF and lung tissue decreased by injecting 2×10~6DCs-SOCS1 on the 1st and 7th day of fumigation(P<0.01);(3)Compared with 1×10~6DCs-SOCS1 goup,IL-21 secretion decreased more obvious in 2x 10~6DCs-SOCS1group on day 1 and day 7(P<0.01);(4)Compared with the control group,there was a significant decrease of IL-21/TGFβin BALF and lung tissues by injecting 1×10~6imDCs、1×10~6DCs-SOCS1 and 2×10~6DCs-SOCS1 on the 1st and 7th day of fumigation(P<0.01);Conclusion:1.imDCs with overexpression of SOCS1 protein can be obtained by lentivirus transfection.2.The level of IL-21 secretion of Th17 was inhibited by imDCs,DCs-SOCS1,and the effect of DCs-SOCS1 was more obvious.3.There is concentration correlation on the inhibition of inflammatory factor release by DCs-SOCS1.4.DCs-SOCS1 could regulate the Th17/Treg imbalance.