Effects And Associated Mechanism Of Overexpression Of ICAM-1Mesenchymal Stem Cells In Experimental Autoimmune Thyroiditis

Objective:1. To investigate the immunoregulatory effect of overexpression Intercellular adhesion moleculesl(ICAM-1) in MSCs (C3H10T1/2-MIGR1-ICAM-1/MSCs) got by our prophase work.2. Experimental autoimmune thyroiditis (EAT) mice modle was carried out in C57BL/6mice using Porcine Thyroglobulin (PTg) and Freund’s adjuvant.3. To study the Therapeutical effect and mechanism of C3H10T1/2-MIGR1-ICAM-1/MSC on EAT.Methods:1. Using LTT and MLR,then assay the numerical value of cpm to detect C3H10T1/2-MIGR1-ICAM-1/MSCs immunologtic function in vitro.2. The6weeks old female C57BL/6mice were immunized with PTg and CFA/IFA at day0and day14to establish the model of EAT. The concentration of TgAb, TPOAb and TMAb in blood serum was tested at day28. Meanwhile, the thyroid tissue were isolated and then embedded in paraffin and stained with hematoxylin and eosin (H&E) for histology examination.3. In mouse EAT model, MSCs were injected into the EAT model mice at the early stage.Mice were separated into6groups named B-MSCs treated group, C3H10T1/2MSCs treated group, C3H10T1/2-MIGR1/MSCs treated group, C3H10T1/2-MIGR1-ICAM-1/MSCs treated group.Different MSCs were injected into the corresponding group at day0,7,14,21. The syno-saline was given in normal control group and EAT model group. At day28, the blood serum and spleen cells were collected to assess the level of TT3、TT4、autoantibodies and the inflammatory factors separately. Thyroid tissue were isolated and then embedded in paraffin and stained with hematoxylin and eosin (H&E) for histology examination.Results:1. Experimental result of LTT and MLR in vitro demonstrated that B-MSCs, C3H10T1/2MSCs, C3H10T1/2-MIGR1/MSCs, C3H10T1/2-MIGR1-ICAM-1/MSCs were capable of suppressing T cells expansion in a dose-dependent manner. There was a dramatic reduction in the T cells proliferation response in overexpression ICAM-1MSCs coculture system comparing to the other MSCs groups.2. Compare with normal control group, thyroid tissue in EAT model group showed inflammatory response and inflammatory infiltration; the level of autoantibodies were significantly increased. These data suggested that EAT mice model was established successfully using PTg and FA.3. We examined the in vivo therapeutical effect of transplanted different MSCs, compare with EAT modle group, thyroid tissue results showed that decreased thyroid follicular damage and invasion of inflammatory cells by MSCs treated groups, C3H10T1/2-MIGR1-ICAM-1/MSCs has obviously inhibitory activity. Comparing with EAT modle group, the level of autoantibodies and IFN-y, IL-1β were significantly decreased in different MSCs treated groups. While the anti-inflammatory cytokine IL-10was increased in different MSCs treated group, Autoantibodies and IFN-y, IL-1β, IL-10had significanter change in C3H10T1/2-MIGR1-ICAM-l/MSCs treated group.Conclusion:1. Experimental result of LTT and MLR in vitro demonstrated that C3H10T1/2-MIGR1-ICAM-1/MSCs was capable of suppressing T cells expansion significantly in a dose-dependent manner.2. EAT mice model was established successfully using PTg and FA.3. C3H10T1/2-MIGRl-ICAM-1/MSCs could enhance the therapeutical effect to EAT and restore the homeostasis compare with other three MSCs treated groups.