The Research On The Role And Mechanism Of Tim-3for The Imbalance Of Th17/Treg In Henoch-Scholein Purpura Patients

BackgroudHenoch-Scholein purpura (HSP) is the most common autoimmune small vessel vasculitis disease for children and adolescents. The pathogen and pathogenesis still remain unclear. The people who are in the situation where their risk of HSP is high are the children and adolesecents among the year of5-15. Adults and infants are not susceptible of HSP. One of the main characterics of this disease is non-thrombocytopenic purpura. The joints, gastrointestinal tract, kidney, brain, lungs may be involved, and scrotum could also be involved.The induced reason of this disease may be infection, vaccination, drug and food. Although HSP is considered to be benigh and selfconfinement, it could affect many organs. Then the disease would delay for a long time before it was cured. The anaphylactic purpura nephritis perhaps caused the patients to take medicine constantly. Some patients perhaps had the kidney failure which would affect the treatment and the patients’living qualities. The replase appeared in one third of the patients. One of the reasons which affect the treatment and the protracted course of this disease is the kidney lesion. So it is impotant to undersdand the pathogenesis, seek a good treatment to cure HSP and enhance the patients’living qualities for the clinical pediatrics doctors. In recent years, some researchers utilize the advanced biology techniques to analyze and explore the pathogenisis of HSP from the molecule and gene lay. Combining amount of epidemiological data, they also want to find the effective cure methods. Although they acquired some achievements, there are still many questions about the pathogenisis and diagnosis to be explicit. It is significant to the diagnosis and prognosis of HSP.Nowadays, some researches have reported the immune dysfunctions of the body were relative to the pathogenesis. Disharmony of T cells, humoral immunologic derangement, diacrisis of the cytokines, abnormal inflammatory response, unusual curor and fibrinolysis pathogenesis and gene susceptibility may be all involved in HSP onset. The basic pathological features are polymorphonuclear cells, lymphocyte cells and eosinophil cells infiltrated aroud the small vessels. IgA compounds, alexin and other immunological compounds deposited on the wall of the small vessels are also the chareactrics of HSP. The serum levels of IgA and some nonspecific proinfiammatory factors such as tumor necrosis factor-a (TNF), interleukin-6(IL-6) raised in the acute HSP acute patients. The above foundings implied inflammation, immunologic derangement and abnormal immune tolerance played an important role for the HSP onset. Sugiyama et al. reported one case of anaphylactic purpura nephritis who was cured by cutting off tonsil and using prednisone. Someya T et al. reported that the patients who are not susceptible for the steroid hormone are remitted effectively by using immunosuppressor. More and more studies indicated that the inflammation spread of HSP could be controlled by immunoregulation.The acknowledeged typical immunologic derangement feature is the antibodies which were produced by B cells clonal activation aggravated the inflammatory response of HSP. But T cells dysfunction and the relative cytokines ratio imbalance were paid more attention by researchers. Some researches have reported that CD4+T cells ratio imbalance and dysfunction may be the important resons for the abnormal immune tolerance of HSP.Thl7cells are the new Th subset which is different with Thl and Th2recently and it is defined expressing-IL-17CD4+T cells. TGF-β and IL-6is the necessary cytokines for the Thl7cells differentiation. IL-23is very important for the amplification and maintain of Th17. Th17cells have played an important role for the onset, development and outcome of the autoimmune disease and chronic inflammation. Nowadays, it has found that the levels of IL-17in serum and tissue enhanced in many autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, sysmetic lupus erythematosus and asthma. Meanwhile, the mice of IL-17deficiency or dealing with the IL-17receptor antagonist resisted the adjuvant induced arthritis and experimental autoimmune celebrospinal meningitis. The above results indicated that IL-17may be involved in the onset of autoimmune disease of human and animal. Many researchers began to focus on the relationship between Th17cells and the pathogenesis of HSP. Many studies found there are Th17cells activation phenomenons in the acute HSP children, and this maybe implied Th17cells involved in the pathogenesis. Li YY el al. reported compared with the healthy control, the frequency of Th17cells and Th2cells in the peripheral blood were higher in acute HSP children. Then they indicated Th17cells were relative with HSP onset closely. Jen HY et al. also studied the similar contents and their results showed that the frequency of Th17cells in peripheral blood elevated and the serum levels of IL-17rised significantly. Then they suggensted Th17cells perhaps take part in the inflammatory responses of HSP. Jun Y el al. found that the expressions of ROR-γt which is the special transcription factor of Th17cells were higher in acute HSP acute patients than in healthy control and their results implied Th17cells existed overactivation phenomenons in acute HSP acute patients.11.-18is a new proinflammatory cytokine which could amply the secretion of IL-6and IL-8,while IL-6and IL-8had the significant correlation with the secretion of IL-17, the differentiation ofTh17cells and the onset of HSP.In the organism, there are many cells which could inhibit and regulate the inflammatory and immune reaction such as dendritic cells,B cells and C4+CD25+regulatory T cells. The regulatory roles of Treg cells are significant. Treg cells play the important roles for the immune tolerance of the organism’ self-antigenic and the resistance of immune reactions. Foxp3gene is not only its special sign, but also is important for the activation and regulatory function of Treg cells. Some reports showed that mice model of deficiency of Foxp3gene could suffer the fatal inflammatory reaction which could be inhibited by transferring Treg cells to the mice model. The mechanism of Treg cells to regulate the immune response belonged amount of types, for example Treg cells could accumulate amonge the DCs to inhibit the activation and propagating of the effector T cells. During the inflammatory and immune response, Treg cells could gather the lesion tissue to play its function. Some Chinese researchers found the abnormal quantity and dysfunction of Treg cells in HSP acute patients recently. Qiang W et al. reported that the frequency of Treg cells in peripheral blood decreased in acute HSP acute patients. Jun Y et al. also found the similar research results. They found that the frequency of Treg cells was significantly lower in acute HSP acute patients than in healthy control. Li YY et al. found the different results which revealed the frequency of Treg cells in perapheral blood had no significant difference between the acute HSP acute patients and the healthy control.In conclusion, although the origin of Treg cells and Thl7cells is CD4+T cells, the imbalance of Thl7/Treg existed in HSP acute patients. The imbalance of Th17/Treg may be related to the HSP onset, the disease severity. It could be the detection index for the HSP severity. It has a great significance to confirm the above opinion for probing the pathogenesis and seeking the effective treatment.Tim-3is newly molecule which could regulate T cells. Through combining with Galectin-9, it could downregulate Thl cells and Th17cells reaction and play the important role in inflammatory and immune response. The mRNA and protein of Tim-3, Galectin-9, T-bet were abnormal in autoimmune diseases patients and animal models such as non-obese diabetes and experimental autoimmune celebrospinal meningitis which all belonged to Thl type diseases. The expression of Tim-3in CD4+T cells became the research hot spot recently. Jones et al. reported the expression of Tim-3in peripheral blood rised in human immunodeficiency virus infectious patients. Ying Liu et al. found the expression of Tim-3mRNA rised in acute rheumatoid arthritis patients. Activating the access of Tim-3/Galectin-9to down-regulate Th17cells had been paid more attention by some researchers. Nowadays, researchers began to keep a watch to the relationship between Tim-3/Th17/Treg and HSP.It has been found that using cytokines or receptor antagonism protein could benefit the gene therapeutic. But the function and the therapeutic effect of the singal cytokine are limited. Focusing on the upstream cytokines and proteins which involved in the pathogenesis became the new research tendacy.In this study, Th17cells, Treg cells, Tim-3and other relative cytokines had been detected in different groups. The aim of this research is to discuss the role of Th17/Treg and Tim-3in the pathogenesis. The results maybe could benefit to making clear the pathenesis of immune tolerance and maintain, expounding the cytokines confusion phenomenon. The results maybe also could provide the new thesis and tactics for the HSP treatment. Objective: The aim of this study is to identify the expression features of Thl7/Treg in peripheral blood in HSP acute patients and the relativity among the expression features of Th17/Treg, clinical manifestations and some laboratory inspections. Another aim of this study is to probe the role and cinical signifieience of Th17/Treg in the pathogenesis of HSP.Methods:23acute HSP acute patients and18healthy controls took part in this study. The frequencies of Thl7and Treg in peripheral blood were detected by How cytometry. The serum levels of IL-17, IL-10and IL-18were examined by enzyme linked immunosorbent assay. The changes of Th17/Treg ratio were analyzed. The relationships among the imbalance of Th l7/Treg, the onset and clinical manifestation were also analyzed. Results:1、The frequency of Th17cells in peripheral blood was higher in HSP acute patients than in healthy controls.2、The frequency of CD3+CD8-IL-21+T cells in peripheral blood was higher in HSP acute patients than in healthy controls.3、The frequency of Treg cells in peripheral blood was lower in HSP acute patients than in healthy controls.4、The ratio of Th17/Treg in peripheral blood was higher in HSP acute patients than in healthy controls.5、The serum levels of IL-17were higher significantly in HSP acute patients than in healthy controls.6、The serum levels of IL-10were lower significantly in acute HSP acute patients than in healthy controls.7、The serum levels of IL-18were higher in acute HSP acute patients than in healthy controls.8、The ratio of Th17/Treg is positive related to the erythrocyte sedimentation rate for acute HSP acute patients.9、The ratio of Th17/Treg is positive related to the numbers of lesion systems for acute HSP acute patients.10、The ratio of Th17/Treg is positive related to the kidney lesions for HSP acute patients.11、The ratio of Th17/Treg has no correlation with the levels of anti-streptolysin O (ASO) and completment3(C3).Conclusions:1、The frequencies of Th17and Treg in peripheral blood were abnormal in acute HSP acute patients. The concentrations of IL-17and IL-10were also unusual in serum in acute HSP acute patients. Thl7cells、Treg cells、IL-17、IL-10and IL-18maybe involve in the pathogenesis.2、The imbalance of Thl7/Treg existed in acute HSP patients. The imbalance of Th17/Treg had significant correlation with the disease severity and kidney lesions. The ratio of Th17/Treg maybe could become the forecast index to measure the HSP severity and kidney lesions. Objective: The aim of this study is to identify the expression of Tim-3in CD4+T cells in MSP acute patients. The correlations among the detected indexes, the clinical manifestation and laboratory inspections were analyzed. The end aim of this part research is to probe the role and clinical significance of Tim-3in the pathogenesis.Methods:18HSP acute patients and15health cases took part in this research. The frequencies of CD4+Tim-3+T cells in peripheral blood were detected by Flow cytometry. The expressions of CD4+Tim-3+Tcells were analyzed. The relationships among the detected indexes, the onset and clinical manifestation were also analyzed.Results:1、The frequencies of CE4+Tim-3+Tcells were lower in acute HSP patients than in healthy controls.2、 The frequencies of CD4+Tim-3+T cells were negative related with ESR in HSP acute patients3、The frequencies of CD4+Tim-3+Tcells had no correlation with rash duration in days, kidney lesions. ASO. C’3and the numbers with lesion systems in acute HSP patients.Conclusions:1、The expression of Tim-3in CD4+T cells in acute HSP acute patients were abnormal. They had correlatDion with the disease severity and some clinical indexes.2、The passage of Tim-3maybe involve the onset of HSP. It perhaps could become the reference index to measuer HSP activity and severity. Objective:The aim of this study is to identify the expression of Thl7+Tim-3+T cells in the peripheral blood in acute HSP patients and to observe the expression of IL-17in peripheral blood mononuclear cells by blocking the road of Tim-3.Methods:15acute patients and10health children took part in this research. The frequencies of Thl7+Tim-3+Tcells in peripheral blood were detected by Flow cytometry. The levels of IL-17cells were detected by enzyme linked immunosorbent assay when the peripheral blood mononuclear cells were dealed with Tim-3monoclonal antibody. The relationships among the detected indexes, the onset and clinical manifestation were also analyzed to identify the role of Tim-3/Thl7/Treg in the pathogenesis.Results:1、The frequencies of Thl7+Tim-3+Tcells were low in healthy controls. It had no expression of Tim-3on Th17cells in the peripheral blood in acute HSP patients.2、Compared with the homotype contrast, the secretion levels of IL-17of the peripheral blood mononuclear cells did not rise by the blocked Tim-3passage in the acute HSP patients.3、Compared with the homotype contrast, the secretion levels of IL-17of the peripheral blood mononuclear cells rised in healthy controls.4、After the Tim-3passage was blocked, the secretion of IL-17of the peripheral blood mononuclear cells in acute HSP patients was higher than it in healthy controls.Conclusions:1、The expression of Tim-3in Th17cells in peripheral blood mononuclear cells in acute HSP acute patients were abnormal. This may be one of the main reasons which leaded Th17cells to be involved in the pathogenisis of HSP.2、During the HSP acute onset, the adjusting roles of Tim-3passsage for Th17cells probably were deficent. If we want to adjust the numbers and function of Th17cells through Tim-3passage in HSP acute onset, we should explore more motheds and consider more causes.