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Objective:To explore the relationship between CD244 and the phenotype and function of CD56bright NK cells of patients with active pulmonary tuberculosis.Methods: PBMCs were isolated from peripheral blood by density gradient centrifugation.The expression of CD244,CD94,NKG2D on the CD56bright NK cells from the active pulmonary tuberculosis patients and healthy controls was detected by flow cytometry.And then analyzed the relationship of the expression of CD244 with Tim3,CD27,CD62L,CCR7,IFN-γ and CD107a in CD56bright NK cells by flow cytometry.Results: The expression of CD244 on the CD56bright NK cells showed no significant difference between the patients with active pulmonary tuberculosis and healthy controls without MTB antigen.The expression of CD244 was significantly increased on CD56bright NK cells of patients with tuberculosis stimulated with MTB antigen.The expression of CD94 and NKG2D on CD56bright NK cells showed no difference between patients and healthy controls.The proportion of Tim3+ cells in CD244+CD56bright NK cells was significantly higher than CD244-CD56bright NK cells.While the expression of CD62L and IFN-γ decreased significantly in CD244+CD56bright NK cells.The expression of CD107a on CD56bright NK cells was not significantly different between CD244+ cells and CD244-cells.Conclusion: The expression of CD244 on CD56bright NK cells in patients with active pulmonary tuberculosis increased significantly,maybe inhibit IFN-γ co-work with Tim3.CD244 has nothing to do with degranulation of CD56bright NK cells.
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Objective To discover T cell reactive peptides of mycobacterium tuberculosis antigen Rv2608 for treatment or adjuvant treatment for tuberculosis.Methods The T cell epitopes of Rv2608 were predicted by bioinformatics.The long-chain peptides were screened with strong affinity with HLA-A0201,HLA-A1101,and HLA-A2402.The physical and chemical characters of the peptides were predicted.The stable and hydrophilic peptides for chemical synthesis were selected.The T cell reactivity of the peptides in patients with active tuberculosis was detected by ELISPOT.The results were compared with the clinical diagnosis kits TSPOT.TB.Results Four peptides of Rv2608 were predicted and screened,and they could be active T cells of patients with active TB.Rv2608p3 was the most efficient to prime the T cell response.Conclusions Software prediction was consistent with the result of ELISPOT.Rv2608p3 might be the candidate for therapeutic vaccine for tuberculosis.
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Objective To investigate the feature of Th1 cytokines induced by Mtb-specific antigen in patients with refractory pulmonary tuberculosis.Methods 28 patients with refractory pulmonary tuberculosis,67 patients with first-treated pulmonary tuberculosis,and 25 healthy controls with positive T.spot (LTBI group) were enrolled.IFN-γ,IL-2 and TNF-α in supernatants from PBMCs stimulated with ESAT-6 and CFP-10 were analyzed with Bender Flowcytomix on flow cytometry.Results The levels of the three cytokines were utmost high in patients with first-treated pulmonary tuberculosis.The lowest level of IFN-γ and IL-2 were induced in patients with refractory pulmonary tuberculosis,and were significantly lower than the LTBI group(Mann-Whitney U ＝ 105.5,162.5,P ＜ 0.01).Conclusions The immunotherapy with IFN-γ and IL-2 may play a role in treatment for refractory pulmonary tuberculosis but not for most of first-treated pulmonary tuberculosis.
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Objective To study the feature of MCP-1 in plasma of active pulmonary tuberculosis patients and the correlation of MCP-1 obtained from PBMCs stimulated with specific Mtb peptides with IFNγ.Methods 20 patients with active pulmonary tuberculosis and 16 healthy controls with positive PPD were enrolled.The concentration of MCP-1 and IFN-γwas analyzed with Bender Flowcytomix on flow cytometry.Results No statistical difference of MCP-1 concentration in plasma was found between TB patients and controls [(263.8 ± 25.31)pg/ml and(212.1 ± 18.04)pg/ml,P ＞ 0.05].But TB patients with continuous respiratory symptoms showed higher MCP-1 in plasma.The concentration of MCP-1 and IFN-γwas significantly elevated in PBMCs culture supernatants.It was significantly higher in TB patients than in controls [(21460 ±3376)pg/ml vs(10910 ±2141)pg/ml,P ＜0.01].There was no correlation between the concentration of MCP-1 and IFN-γ.Conclusions The concentration of MCP-1 in plasma may be related to the progress of the pulmonary tuberculosis.MCP-1 stimulated by Mtb-specific peptides may be one of the biomarkers for TB diagnosis.
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Objective To study the feature of MIG and IFN-γ obtained from PBMCs stimulated with Mtb specific antigens and the potential value in the differential diagnosis of active pulmonary tuberculosis from bacterial pneumonia and primary lung cancer. Methods 90 patients with active pulmonary tuberculosis and 31 patients with bacterial pneumonia and primary lung cancer were enrolled. MIG and IFN-γin supernatants from PBMCs stimulated with Mycobacterium tuberculosis-specific antigens were analyzed with Bender Flowcytomix on flow cytometry. The diagnostic values were established based on receiver operating characteristic curve analysis. Results PBMCs stimulated with Mtb-specific antigens produced significantly higher levels of MIG compared with IFN-γ The level of MIG in active pulmonary TB patients was significantly higher than in controls(3023.0 pg/ml vs 112.5 pg/ml, P ＜0.0001). The MIG and IFN-γtests were positive in 96. 8 and 86. 7% of the TB patients, the specificity was up to 94. 4 and 87. 1%. With combination of MIG and IFN-γtests, the positive rate increased among TB patients to 97. 8% without a significant decrease in specificity. Conclusions The responses of the MIG and IFN-γagainst to Mtb-specific antigens could be used to discriminate newly-treated active pulmonary tuberculosis fiom bacterial pneumonia and primary lung cancer. Combination of MIG and IFN-γ might be a simple and quick approach to diagnosis newly-treated active pulmonary tuberculosis.