The Immune Regulation Effects Of AntagomiR-155on EAMG Mice

Objective:We established the EAMG model to observe the therapeutic effects of antagomiR-155on the antibody secreting and activation of B cell; to explore the effects and mechnism of antagomiR-155in EAMGMethods:C57BL/6mice (6-8week) were immunized with the fragment97-116of T-AchR to establish the animal model of EAMG. The immunization were repeated on day30and day60,then the mice were evaluated by clinical score of EAMG on day70.The mice were then divided into three groups, including PBS group、Control group and antagomiR155group. The control group were treated with the nonsesce fragments, antagomiR-155group were treated with antagomiR-155. On the5th day after the treatments, blood and spleen of the mice were collected. T cells and B cells were obtained by immunomagnetic beads,then the proliferation of cells marked with fluorescent dye CFSE were detected by folw cytometry. Also flow cytometry was used to identify the ratio of memory B cells and plasma B cells, also the expression of CD40、CD80、CD86on the surface of B cells. ELISA was used to dected AchR-specific antibodies total IgG and it’s isotypes. MiRNA qRT-PCR was used to detect the expression of transcription factors p38MAPK and C/EBPβ.Resrults:1.Comparing with the PBS group and control group,the levels of antibody of IgG and IgG1in blood were significantly reduced in antagomiR-155group (P<0.001), but there was no sigificant difference in the levels of IgG2a、IgG2b、IgG2、IgG3in all three groups(P>0.05).2. The ratio of plasma B, especially memory B cells cells were reduced in antagomiR-155group, comparing with the PBS group and control group(P=0.029,P<0.001). There was no sigificant difference in the ration of memory B cells and plasma B cells between BPS groups and control group.3. Comparing with the PBS group and control group,the expression of CD40and CD86were increased significantly in antagomiR-155group. There was no significant difference in the expression of CD80among each group.4. B cell proliferation was be impaired after stimulated by AchR and R97-116in antagomiR-155group than that of PBS group and control group,and the difference was obvious statistical significant. The proliferation of B cells were happended after the stimulation of ConA and there was no significant difference among each group. No proliferation of B cells were happended when them was treated with serum,also there was no significant difference among each group. 5. Comparing with the PBS group and control group,the miRNA expression of p38MAPK were reduced significantly in antagomiR-155group (P<0.001). There was no significant difference among each group on the expression of C/EBPβ(P>0.05)Conclusion:AntagomiR-155could regulate the immunological system of EAMG; it was involved and play a negative role in the process of B cell proliferation and antibody production。...