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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 compare the mRNA expression of IRG1in PBMCs from patients with TB, individual with latent infection and healthy controls, and investigate its diagnostic value for Mycobacterium tuberculosis infection.Methods: The mRNA expression of IRG1 from PBMCs stimulated with Mtb-specific antigens was quantitatively detected by quantitative real-time PCR ( qPCR).Receiver-operating-characteristic ( ROC) curves were used to determine the cutoff points yielding the highest specificity and sensitivity,and discriminative ability was evaluated by the area under the ROC curve.Results: The mRNA expression of IRG1 in healthy controls was significantly lower than that in patients with tuberculosis and LTBI ( P0.01536 ) to identify Mycobacterium tuberculosis infection was 0.91, with 76.47% sensitivity, 96.30% specificity, LR 20.65,and 85.2% of cases correctly classified.Conclusion: IRG1 may be used as a biomarker for diagnosing Mycobacterium tuberculosis infection.
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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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SELEX is a newly developed biochemical technique,which filter out high specificity and high affinity ligand for the target molecules through the identification of aptamer combined with the target molecules.The specific aptamer was used in a variety of clinical applications,such as diagnosis of the disease,development of new therapeutic drugs and even directly applied to disease treatment.
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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.
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Objective To study the expression of CD27 and CD28 in antigen-specific CD4~+T cells in patients with pulmonary tuberculosis and healthy people, and understand the role of differentiated stages of CD4~+T cells in the pathogenesis of tuberculosis. Methods The expression of CD27 and CD28 was analyzed by CD4, CD154, CD27 and CD28 staining and flow cytometry. The distributions of CD27 and CD28 in antigen-specific CD4~+T cells were compared between patients with pulmonary tuberculosis and healthy controls. Results In patients of pulmonary tuberculosis, the frequencies of CD27 + CD28 + (early differentiated stage), CD27~- CD28~+ and CD27~+ CD28~- (intermediate differentiated stage), CD27~- CD28~-(fully differentiated stage) T cell subsets in antigen specific CD4~+T cells were (49. 55 ±6. 15)%, (26. 85 ±3. 87)% ,(7. 2 ± 1.37)% and ( 16. 35 ±3.97)%, respectively. In healthy controls, the frequencies of the four subsets in antigen-specific CD4~+T cells were ( 51.81 ± 4. 94 ) %, ( 29. 83 ± 5.33 ) %, ( 12. 65 ±4. 48)% and (5.7±2)%, respectively. The early differentiated CD4~+T cell was the major subset both in patients and healthy people, however, which had significant difference compared with the fully differentiated subset ( t = 2. 26, P < 0. 05 ). Conclusion The population frequency of the fully differentiated CD4~+T cells in patients with pulmonary tuberculosis was significantly higher than that in healthy people. This suggested that the differentiation degree of the antigen-specific CD4~+T cell might be related with pulmonary tuberculosis.
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Objective To study population frequencies of CD4+,CD154+ T cell subset in patients with pulmonary tuberculosis and controls with positive PPD reaction. Methods Flow cytometry was used to detect the CD4+,CD154+ T cell subset, the population frequen-cies in patients with pulmonary tuberculosis and controls were compared. Results The expression level of CD154 was higher when PE-la-beled CD154 antibody was added during stimulation period, compared with CD154 labeling after stimulation(1.51±0. 36/0. 40±0. 13, P <0.05). The CD154+ cells were not detectable in fresh isolated CD4+ T cells, but significantly increased after stimulation with specific anti-gens. The population of CD4+, CD154+ T cell subset was significantly reduced in patients with active pulmonary tuberculosis, compared with healthy controls with PPD positive reaction(0. 72±0. 32/1.65±0. 76, P <0. 01). Conclusions The population of CD4± ,CD154± T cell subset was significantly reduced in patients with active pulmonary tuberculosis, which indicated that it may play an important role in the de-velopment of tuberculosis.
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Objective To study the genes modulated during plasma cell development and their regulating network. Methods The changes of gene expression during differentiation from mature B cells to plasma cells were studied using cDNA microarray and RT PCR. Results The gene expression profile during plasma cell differentiation was established. During mature B cell plasma cell differentiation, numerous genes were modulated, including genes involving immunity, cell growth, survival, and signal transduction. The expressions of several molecules in the B cell antigen receptor (BCR) signaling pathways, such as Ig?, Ig?, Lyn, Syk, BLNK, SWAP 70, and SHP 1 were down regulated. The decreased expression of some of these molecules might be associated with the down regulation of the transcription factor BSAP. Conclusion During plasma cell differentiation, molecules involving BCR signaling pathways are down regulated.