Objective:To investigate the specific stimulation of the B cell epitope peptides of Mycobacterium tuberculosis antigen (Mtb-Ag) on human peripheral γδ T cell proliferation.Methods: We selected the sequences of B cell epitope peptide from Mtb-Ag that were reported in literature and T cell epitope peptide that recently identified in this laboratory to synthesize six peptides of B cell epitopes (BP1-BP6) and two peptides of γδ T cell epitopes (TP14,TP15).The 24-well culture plates were coated with these peptides.The PBMCs were isolated from peripheral blood of healthy individuals and stained with CFSE,followed cultured for 12 days in the IL-2 containing medium.Mtb heat resistant antigen ( Mtb-HAg ) group as positive control and IL-2 only group as negative control.The percentages and proliferation index of γδ T cells were determined by flow cytometry.Results: By using Wilcoxon signed rank test for paired comparison of negative control group,the percentages of γδ T cells in cultured PBMCs with BP2,BP4 and TP14, TP15 and Mtb-HAg increased significantly in 14 samples (P<0.05);and the proliferation index of γδT cell in cultured PBMCs with BP2,BP4,BP5,BP6 and TP14,TP15 increased significantly in 7 samples (P<0.05).Conclusion: Taken together,the B cell epitope peptides from Mtb Antigens are capable of stimulating the γδ T cell proliferation specifically in vitro.Although there was individual difference inγδT cell proliferative response to B cell epitope peptides,these results strongly suggest the B cell epitope peptides also can specifically trigger the TCR ofγδT cells.

Cultivation of murine thymic epithelium has been established.Murine thymic epi-thelial cultural supernatant showed activity increasing both spontaneous ~3H-TdR inco-rporation and ConA-stimulated proliferation of thymocytes by the exponent of 1.5-2at dilution 1:8 to 1:32,the peak of the activity was present at 10-20 or 15 days ofcultivation.However,the thymic epithelial supernatant from cultures at 6-7 daysshowed activity suppressing both proliferations of thymocytes at dilution 1:8-1:16.Theresult showed thymic epithelial supernatant had no 1L-2 activity.

Objective To investigate the clinical significance of video-assisted thoracoscopic thymectomy. Methods Eighteen patients with thymic diseases or myasthenia gravis (MG) received video-assisted thoracoscopic surgery (VATS) of either total thymectomy (7 patients) or extended thymectomy (11 patients with MG) from September 2002 to June 2004. Results Thoracoscopic thymectomy was accomplished in 17 patients and a conversion to open mini-incision (7 cm) operation was needed in 1 patient. Postoperative temporary mechanical ventilation (

Objective To investigate the proliferative response of peripheral blood mononuclear cells(PBMC) and its depressant effect on small cell lung cancer(SCLC)cells(H446) in patients with paraneoplastic neurological syndrome(PNS).Methods PBMC of 7 patients with PNS and 6 patients with SCLC were cultured with interleukin(IL)-2 in vitro,then cultured separately or mixed with H446 respectively.The proliferation index(PI,the ratio of cellular score which had proliferated and that had not proliferated) of H446,PBMC,CD4+,CD8+ T cell and the ratio of CD4+,CD8+,CD4+/CD8+ were analyzed with flow cytometry and compared with normal control group.Results Compared with cultured separately,the PI of H446,PBMC,CD4+,CD8+T cell in PBMC of PNS and SCLC groups cultured with H446 were not significantly different.Stimulated by IL-2,the ratio of CD4+ T cell in PNS patients [(76.54 ? 3.96)%]was higher than that in normal control group[(51.75 ? 17.3)%](P

Objective:To investigate involvement of the Ras/Erk signal pathway in activation of human ??T cells stimulated by Mycobacterium tuberculosis low molecular peptide antigen(Mtb-Ag).Methods:PBMC were isolated from peripheral blood of healthy subjects and simulated in vitro with Mtb-Ag,CD3mAb or PMA and inomysin(IM).CD69 expression of total T cells and ??T cells were measured by flow cytometry at different hours after stimulation, and the number of expanded ??T cells were counted after 10 days of culture. PD98059,a specific inhibitor for Erk pathway was used to treat PBMC for 30 min before stimulation.Results:The proportions of CD69 expressing cells in total T cells and ??T cells were same as 99% at 6 h after stimulation of PBMC with PMA +IM and similar about 55% at 24 h after stimulation with CD3 mAb,whereas those at 24 h stimulation of PBMC with Mtb-Ag were 75.2% and 16.0%,respectively.CD69 expression and proliferation of ??T cells activated by Mtb-Ag were markedly inhibited by pretreatment of PBMC with PD98059.Conclusion:Mtb-Ag is a specific stimulator for activation of ??T cells, and Ras/Erk signal transduction pathway involves in the activation of ??T cells.

0.05),but the proliferation rate(P

Objective:To establish the model of apoptosis of activated human ??T cells induced by restimulating with Mycobacterium tuberculosis antigen(Mtb-Ag). To investigate the roles of p38 and phosphatidylinositol 3 kinase(PI3K) pathways in the apoptosis of activated human ??T cells induced by restimualting wih Mtb-Ag.Methods:Mtb-Ag activated human T cells(MtbAT) were cultured for 15 days to 25 days and restimulated with three concentrations of Mtb-Ag for 24 hours, and the apoptosis of ??T cells were measured by flowcytometry(FCM) using Annexin-V-FITC/PI staining. Mtb-AT were restimulating with Mtb-Ag(10 ?g/ml) for 3, 6, 12 and 24 hours, the apoptosis of ??T cells were detected. Mtb-AT cells were pretreated with SB203580(an inhibitor for p38 pathway), or LY294002(an inhibitor for PI3K pathway) for 60 minutes, and restimulating with Mtb-Ag for 3 hours, the apoptosis of ??T cells were detected.Results:Both 10.0 and 20.0 ?g/ml Mtb-Ag significantly induced the apoptosis of ??T cells(P0.05). Compared with control, the apoptosis of ??T cells could be significantly induced by restimulating MtbAT with Mtb-Ag(10.0 ?g/ml) for 3, 6, 12 and 24 hours(P0.05) in the percentages of apoptosis of ??T cells restimulated by Mtb-Ag(10.0 ?g/ml) between for 3 hours and for 24 hours, the percentages of apoptosis of the latter is higher than the former about 7.55%. The apoptosis of ??T cells induced by restimualting wih Mtb-Ag could be inhibited by SB203580(80.0 ?mol/L) or LY294002(10.0 ?mol/L), the inhibition rate of apoptosis was 91.6% and 43.1%, respectively.Conclusion:We established the model of apoptosis of activated human ??T cells by means of using Mtb-Ag(10.0 ?g/ml) to restimulate activated ??T cells for 3 hours. The test of inhibitors of signalling molecule suggested the signalling pathways including p38 and PI3K, participated in the apoptosis of activated human ??T cells restimulated by Mtb-Ag.

Objective To observe the role of human hepatocarcinoma HepG2 cell culture supernatant on T lymphocyte of the activation, proliferation, IL-2 secretion and inducing apoptosis in vitro. To explore the mechanism of immune escape in tumor.Methods Peripheral blood mononuclear cells (PBMC) were cultured with different concentration of tumor cell culture supernatant. Cell proliferation was examined with standard cytometry and MTT. The apoptotic rate and immune phenotype were analyzed by FACS. The level of IL-2 was determined with ELISA.Results Adding HepG2 supernatant, there was no significant influence in activation of lymphocyte, but the proliferation rat and the secretion of IL-2 were obviously decreased. The lymphocyte apoptosis rate was raised.Conclusion The tumor cell may produce a certain soluble protein, thus escape from the immune surveillance of tumor-bearing host.

Objective: To study the antibacterial activity in vitro of Sanhuang Shaoshangling on several common bacteria after changing the preparation process. Methods:Using the new technology of ethanol reflux extraction and water extraction, Sanhuang Sha-oshangling was prepared. A micro-broth dilution method was used to determine the minimum inhibitory concentration ( MIC) of San-huang Shaoshangling for escherichia coli, pseudomonas aeruginosa, staphylococcus aureus and type B paratyphoid salmonella after chan-ging the preparation process. The antibacterial effect of Sanhuang Shaoshangling prepared by the new and the old technology was com-pared, and the inhibition zone was detected by a paper strip method. Results:The antibacterial effect of Sanhuang Shaoshangling pre-pared by the new technology on escherichia coli, pseudomonas aeruginosa, staphylococcus aureus and type B paratyphoid salmonella was better than that of the products prepared by the old technology with the minimum inhibitory concentration ( MIC) of 16 μg·ml-1 , 16μg·ml-1 , 8 μg·ml-1 and 16 μg·ml-1 , respectively, while that of the product prepared by the old technology was 32 μg·ml-1 , 64 μg·ml-1 , 16 μg·ml-1 and 64 μg·ml-1 , respectively. Conclusion: The antibacterial effect of Sanhuang Shaoshangling after changing the preparation process is more significant, which shows certain clinical significance.

Objective:To explore the role of CD28 in the activation of human peripheral blood ?? +T cells by Mycobacterium tuberculosis (Mtb) low molecular peptide antigen in vitro Methods:Mtb antigen and anti CD28 monoclonal antibody (mAb) were used as signal 2 to stimulate the r? + T cell from PBMC the expression of CD28 molecule on ?? +T cells, proliferation rate of ?? +T cells and expression of CD69 molecule on activated ?? +T cells were analyzed by using flow cytometry Results:CD28 molecule was expressed on 50% of ?? +T cells Neither Anti CD28 nor Mtb antigen alone, but both presence, could stimulate ?? +T cells activation and proliferation CD69 molecule was expressed on activated ?? + T cells.Conclusion:The CD28 molecule could provide the costimulatory signal in the full activation of ?? +T cells by Mtb low molecular peptide antigen CD69 molecule was also an early activation marker of ?? +T cells