RESUMEN
ObjectiveTo investigate the effect of Yiqi Jiedu prescription-containing serum on the proliferation of medullary thymic epithelial cells (mTEC) and regulatory T (Treg) cells in myasthenia gravis (MG) patients with thymus hyperplasia. MethodAccording to serological methods,35 SD rats were adaptively fed for one week and randomized into the low-,medium-, and high-dose Yiqi Jiedu prescription groups,control group, and prednisone group,with seven rats in each group, which were then gavaged with the corresponding drugs for one week for preparing the drug-containing serum. The effect of Yiqi Jiedu prescription-containing serum at different concentrations on the proliferation of mTEC and Treg cells were determined by cell counting kit-8 (CCK-8) assay. Besides, the effect of mTEC and Yiqi Jiedu prescription-containing serum on Treg cell proliferation were observed through co-culture. ResultThymocytes were cultured for a period of time. Their mean positive rate revealed by flow cytometry using mTEC characteristic marker Ulex europaeus agglutinin Ⅰ (UEAI) was 92.54%. Treg cells were sorted by magnetic beads. The purity of Treg cells after repeated magnetic bead sorting was as high as 92%. mTEC and Treg cells showed high positive expression rates,and their cell purity met the requirements of subsequent experiments. When the concentration of Yiqi Jiedu prescription-containing serum was 2.5%-15%,it exhibited an inhibitory effect against mTEC and Treg cells. When the concentration was equal to or greater than 20%,it promoted cell proliferation,which was further enhanced with the extension of action time. The results after 48 h of culture showed that compared with the control group,prednisone and low-dose Yiqi Jiedu prescription had no significant effect on the proliferation of these two kinds of cells,but the medium- and high-dose Yiqi Jiedu prescription remarkably reduced their proliferation inhibition rate (P<0.01). After co-culture with mTEC, the control group was not significantly different from the prednisone group and the low-dose Yiqi Jiedu prescription-containing serum group in the proliferation of Treg cells,while the medium- and high-dose Yiqi Jiedu prescription-containing serum groups significantly lowered the proliferation inhibition rate (P<0.01). ConclusionYiqi Jiedu prescription-containing serum affects the proliferation of mTEC and Treg cells in MG patients with thymus hyperplasia. Compared with the solely cultured Treg cells isolated from MG patients,the Treg cells co-cultured with mTEC exhibit enhanced proliferation in MG patients,suggesting that mTEC can regulate the proliferation of Treg cells. This effect becomes more obvious after the intervention with Yiqi Jiedu prescription-containing serum,indicating that intervention effect of Yiqi Jiedu prescription on Treg cells can be produced during its treatment of mTEC, which may be one of the mechanisms of Yiqi Jiedu prescription-containing serum in alleviating MG.
RESUMEN
Thyroid-associated ophthalmopathy(TAO)is an organ-specific autoimmune disease, which will cause a series of symptoms to significantly reduce the health level and life quality of patients. The pathogenesis of TAO has not been fully clarified. At present, there is a lack of unified and mature treatment scheme of it. Indeed, T-helper 17 lymphocyte(Th17)cells, regulatory T(Treg)cells and their imbalance are closely related to the immunological pathogenesis of TAO. It is currently believed that the cytokines secreted by Th17 cells can not only promote the inflammatory response of TAO and the fibrosis of orbital connective tissue, but also inhibit the adipogenic differentiation of TAO orbital connective tissue. In addition, Treg cells mainly exert immunosuppressive effect on TAO and delay the disease progression. At the same time, there is a dynamic balance relationship between Th17 and Treg cells, the imbalance of Th17/Treg cells can trigger the occurrence and development of TAO. This paper mainly expounds the influence mechanism of Th17, Treg cells and their balance on TAO, and analyzes the reasons for the differences between different research results, so as to provide some reference for the study of the pathogenesis and clinical treatment of TAO.
RESUMEN
Objective By using mouse orthotopic lung transplant model, we investigated the immune mechanisms of an-ti-CD3 induced lung allograft protection .Our study intends to further dissect the features of lung transplant immunology and to provide a novel therapeutic insight for the clinical application of anti-CD3 mAbs after lung transplantation.Methods Murine orthotopic allogeneic lung transplants were performed in C57BL/6 wild type(WT) mice using major histocompatibility complex (MHC) fully mismatched BALB/c donors.Syngeneic transplants were also performed in WT C57BL/6 mice using C57BL/6 donors.For immunosuppressive therapy, allograft recipients received 50g dose of anti-CD3 by intraperitoneal injection on days 2, 3, 4, 5, 6 and 9 post-operation(n=4).At day 10, histopathologic characteristics and rejection status of the pulmonary grafts were assessed.The severity of acute rejection was graded by the pathological score , and T cell and neutrophil infiltration in the pulmonary grafts was evaluated by immunohistochemical(IHC) staining for CD3 and myeloperoxidase(MPO) respective-ly.Real-time RT-PCR was performed for FoxP3, IL-17A and IFN-γexpression in the pulmonary grafts.The percentage of FoxP3+Treg in total CD4+T lymphocytes from the recipient spleens was analyzed by FACS.Results 10 days after transplan-tation, histopathologic examination demonstrated that there is no apparent acute rejection observed in the pulmonary isografts , whereas allografts from untreated recipients have marked inflammatory cell infiltration and pulmonary parenchyma lesion .IHC staining for CD3 and MPO showed that the allograft-infiltrating cells of perivascular layers are mainly T lymphocytes , and the cells around the small airways are mostly neutrophils .Anti-CD3 treatment significantly alleviated the acute rejection of pulmo-nary allografts, when compared with the untreated group.Real-time RT-PCR showed that the expression levels of IL-17A and IFN-γin allografts were markedly elevated compared to those in isografts, and anti-CD3 increased the expression of FoxP3, and reduced the expression of IL-17A and IFN-γin the pulmonary allografts.FACS analysis of splenocytes showed that the percent-age of Treg in total CD4+T lymphocytes increased significantly in the anti-CD3 treated allograft recipients, as compared with the isograft and untreated allograft recipients.Conclusion Anti-CD3 mAbs may alleviate acute rejection of the pulmonary al-lografts by promoting FoxP3 expression and Treg development.