The Immune Effect Of Co-Inhibitory Molecule Tim-3/Galectin-9 In Pulmonary Tuberculosis

Objective:To study the changes of negative co-stimulatory molecule Tim-3/Galectin-9 and related cytokines in pulmonary tuberculosis,and to explore its immune regulation in the development and progression of tuberculosis.Method:This study included sixty-five patients who were diagnosed as pulmonary tuberculosis?PTB?,fifty-eight persons as healthy control?HC?.Collection the peripheral blood samples of healthy control group and pulmonary tuberculosis group for analysis by flow cytometry and real-time polymerase chain reaction?qRT-PCR?.Serum sample was for cytokine measurement by CBA and ELISA.The lung tissue samples collection used for immunohistochemistry analysis.?1?Flow cytometry was used to detect the percentage of Tim-3 on CD4⁺CD25⁺CD127^lowTreg in Peripheral blood.?2?Immuhistochemistry was used to observe the expression of Tim-3,Galectin-9 and Foxp3 in lung tissue.?3?qRT-PCR was used to detect the expression of Tim-3,Galectin-9 and related cytokines Foxp3,TGF-?,IL-10,IFN-?and TNF-?.?4?Cytometry Beads Assay was used to detect the serum levels of TGF-?,IL-10,IFN-?,TNF-?,IL-2 and IL-4.?5?ELISA was used to detect the serum levels of Galectin-9 and sTim-3.?6?The correlations between Tim-3,Galectin-9,Foxp3,TGF-?,IL-10,IFN-?and TNF-?of patients with pulmonary tuberculosis were analyzed by Pearson correlation test.?7?After Galectin-9 stimulation of Tim-3/Galectin-9 pathway in vitro,CBA detect the levels of IL-10 and TGF-?.?8?After anti-Tim-3 blockade Tim-3/Galectin-9 pathway in vitro,CBA detect the levels of IL-10and TGF-?.Result:?1?Flow cytometry results shown that compared with healthy control group,the ratio of Tim-3⁺CD4⁺CD25⁺CD127^lowT cells was significantly higher than that in healthy control group?P<0.01?.?2?Immuhistochemistry results shown that the expression of Tim-3,Galectin-9 and Foxp3 in pulmonary tuberculosis group was significantly higher than in healthy control group?P<0.01?.?3?Results of qRT-PCR demonstrated that,compared with healthy control group,the expression of Tim-3,Galectin-9,Foxp3,TGF-?and IL-10 mRNA in pulmonary tuberculosis group were significantly increased?P<0.05?,the expression of IFN-?and TNF-?mRNA decreased in pulmonary tuberculosis group compared with healthy control group?P<0.01?.?4?CBA results shown that compared with healthy control group,the serum levels of IL-10,TGF-?and IL-4 were significantly increased in pulmonary tuberculosis group?P<0.01?,the serum levels of IFN-?,TNF-?and IL-2 were decreased obviously in pulmonary tuberculosis group?P<0.05?.?5?ELISA results shown that compared with healthy control group,the serum level of Galectin-9 and sTim-3 was increased obviously in pulmonary tuberculosis group?P<0.01?.?6?Pearson analysis showed that there was a positively correlation between Tim-3,Galectin-9 and Foxp3,TGF-?,IL-10,while between Tim-3,Galectin-9 and IFN-?,TNF-?were negative correlated?P<0.05?.?7?According to the clinical characteristics of pulmonary tuberculosis patients,the levels of TGF-?in the cavitary group were significantly higher than those in the no-cavitary group?P<0.05?,the levels of IL-10 in the retreat group were significantly higher than those in the initial group?P<0.05?.?8?After Galectin-9 stimulation of Tim-3/Galectin-9 pathway,CBA results shown that the levels of IL-10 and TGF-?were significantly increased?P<0.01?.?9?Results of anti-Tim-3 blockade of Tim-3/Galectin-9 pathway,the levels of IL-10 and TGF-?were decreased obviously?P<0.05?.Conclusion:This study found that in the pulmonary tuberculosis group the expressions of negative costimulatory molecules Tim-3,Galectin-9,Tim-3⁺CD4⁺CD25⁺CD127^lowT cells and related immune moiecule?Foxp3,TGF-?and IL-10?were increased,Th1 cells related immune moiecule?IFN-?,TNF-?and IL-2?were decreased.Activate or block Tim-3/Galectin-9 pathway in vitro experiment,will influence the level of TGF-?and IL-10.It is suggested that Tim-3/Galectin-9 in pulmonary tuberculosis patients may promote the inhibitory effect of Treg cells,which is the reason of remove mycobacterium tuberculosis is difficult in vivo and mycobacterium tuberculosis escape the immune response.