The Interaction Of Chemokine CCL20 And Th17 Cells In The Pathogenesis Of IgANephropathy

Objective:Detection of the expression level of chemokines CCL20 in patients with IgAN and the expression of its receptor CCR6 in peripheral immune cells, and further exploration the interaction between CCL20/CCR6 and Th17 cells in IgAN,in order toprovide an excitong new way for the pathogenesis of IgAN.Methods:1、Incubating human glomerular mesangial cells with IgAN patients serum for 24 hours, using RT-PCR technology to screen high expression of chemokines.Additonaly, using Jacalin-Agrose affinity chromatography to extract the Ig A1 in IgAN patient serum and healthy people’s, then respectively incubated with HMC for 24 hours, and then detected of two groups of CCL20 m RNA expression level by RT-PCR.Furthermore,we used the method of ELISA to detect serum chemokine expression level in the two groups of IgAN and healthy people.2、Using transwell chemokines experiment and flow cytometry analysis to analyze the expression level of the chemotaxis of CCL20 cells and its receptor in what kind of cells.3、Usingdyeing intracellular factor experiments and cytokine expression spectrum experiment to detect the relationship between CCR6~+ cells and Th17 cells.4 、 Application of immunohistochemistry and immunofluorescence technique to respectively detect the expression of CCL20, T cells and Th17 cells in the kidneys of IgAN patients, other kidney disease tissue and tissue adjacent to carcinoma.Results:1、The chemokine of CCL20 expression increased after we incubated HMC with IgAN patients serum.In addition, both the two groups of serum Ig A1 incubation with HMC, and singly detectedthe serum of IgAN patients and healthy controls, the results were the expression of CCL20 levels in patients with IgAN group wassignificantly higher than healthy control group.2、Chemokines CCL20 was priority for chemotaxis peripheral blood T cell(mature T cells),and its receptor CCR6 expression level and the average fluorescence intensity were higher in mature T cells than na?ve T cells.3、CCR6~+ T cells wrer mainly Th17 cells which secretion of IL-17, and these Th17 cells secreted a large number of inflammatory cytokines to cause kidney damage or further damage.4、Using immunohistochemical method,we can detected CCL20 in patients with IgAN kidney organization, while not in control group;Using immunofluorescence method, we can also detected CD3~+CCR6~+IL-17~+ cells, but not in the control group.Conclusion:In summary, high expression of CCL20 in IgAN cause CCR6~+ T cells(mainly Th17 cells) high expression level,and then Th17 cells secreted lots of inflammatory factors and cytokines cause kidney further damage. Thus, block CCL20/CCR6 could become a new therapeutic method for IgAN.