Objective: This study analyzed the T lymphocytes and Th1/Th2 type cytokine profile shift in the peripheral blood ofpatients with recurrent genital herpes (RGH). Methods: Immunofluorescent staining or cell surface anti...Objective: This study analyzed the T lymphocytes and Th1/Th2 type cytokine profile shift in the peripheral blood ofpatients with recurrent genital herpes (RGH). Methods: Immunofluorescent staining or cell surface antigenand intracellular cytokines(IL-2、IL-4、IL-12、IFN-r)inperipheral blood from 20 RGH patients and 10 controls wereanalyzed using flow cytometric techniques. Results: RGH patients had signiflcantly lower levels ofCD3^+T cells, CD4^+T cells and CD4^+ T/ CD8^+ T cells ratiocompared to control levels (P<0.001), and IL-2-producing,IFN-r-producing and IL-12-producing T cells were increasedin RGH patients (CD4^+T: P<0.001, CD8^+T: P<0.05respectively), whereas IL-4-producing T cells were increased inRGH patients compared to controls (CD4^+T: P<0.05; CD8^+T:P<0.001 respectively). Conclusions: RGH patients have T lymphocyte subsetvariations and Th1/Th2 cytokine changes. The increase in Th2cells Th1/Th2 imbalance may have important implications forRGH pathogenesis.展开更多
Objective： To evaluate whether radiofrequency ablation （RFA） might have an influence on immune status in hepatocellular carcinoma （HCC） patients. Methods： We measured the T lymphocytes, B lymphocyte and NK cell...Objective： To evaluate whether radiofrequency ablation （RFA） might have an influence on immune status in hepatocellular carcinoma （HCC） patients. Methods： We measured the T lymphocytes, B lymphocyte and NK cells, and determined the population of Thl, Th2, Tcl and Tc2 of peripheral blood samples taken from 26 HCC patients before and after RFA. Results： The proportion of Typel cells （Thl and Tcl） and NK cells were significantly increased after RFA, especially in patients of the following subgroups： male, age〉55 years, pathological grade Ⅰ-Ⅱ tumor, clinical stage Ⅰ-Ⅱ or Child-Pugh A and B. Conclusion： TypeⅠ cells and NK cells in HCC patients were increased in a short period after RFA.展开更多
To investigate the effects of overall alkali of a traditional Chinese medicine “Tongbiling” (brucine and strychnine alkaloids in main) on the cytokines expression in Th1 and Th2 cells in the synovial fluid of patien...To investigate the effects of overall alkali of a traditional Chinese medicine “Tongbiling” (brucine and strychnine alkaloids in main) on the cytokines expression in Th1 and Th2 cells in the synovial fluid of patients with rheumatism arthritis and their signal pathway, the mononuclear cells in the synovial fluid (SFMC) of patients were isolated by Ficoll-Hypaque gradient centrifugation, and the CD3 + CD69 + and CD3 + HLA-DR antigen were analyzed by flow cytometry in comparison with those of the peripheral blood. The rest of cells were cultured after resuspension with RPMI 1640 culture medium. Phorbol 12,13-dibutyrate (PDB) and ionomycin were added successively into the culture with various concentration of overall alkali Tongbiling (TBL). After 4 h of cultivation, the expression of IFN-γ and IL-4 in CD3 + cells were analyzed by flow cytometry. The influence of overall alkali TBL (100?mg/L) on the intracellular calcium was investigated after Fluo-3/AM labeling and stimulation with PDB and ionomycin at 1, 2, 4 and 10?min, and the influence of TBL on the expression of CD3 + CD69 + cells were determined with stimulation of PDB for 24?h in the whole blood lymphocytes culture. It was found that the percentage of T cells bearing CD69 was significantly up-regulated (77%), while that of T cells bearing HLA-DR was 44% in the synovial mononucleated cells. After PDB and ionomycin stimulation, the expression of IFN-γ in CD3 + cells were up-regulated, but there was no change on the expression of IL-4 in CD3 + cells, indicating that ratio of Th1/Th2 was significantly increased and Th cells differentiate to Th1 cells in mainly. Four concentrations of overall alkaloid of TBL (200?mg/L, 100?mg/L, 50?mg/L, 25?mg/L) could down-regulated the expression of IFN-γ in CD3 + cells and the Th1/Th2 ratio obviously, but all the concentrations of the overall alkaloids had no effect on the expression of IL-4 in CD3 + cells. 100?mg/L concentration of the overall alkaloid did not down-regulate the intracellular calcium level. Each concentration of the overall alkaloid could down-regulated the expression CD69 obviously on the PDB-activated mouse T cells. It concluded from the above observations that the overall alkaloid of TBL could relieve the inflammatory and immune damages by suppressing the expression of Th1 type cytokines and Th1 cell differentiation, regulating the imbalance of Th1/Th2 cells and inhibiting the early activation of the T lymphocytes bearing CD69. There was no remarkable influence on the intracellular calcium signaling transduction pathway. The inhibitory effected on T cells to express IFN-γ might be due to the suppression of PKC-MAPK signaling pathway. From the standpoint of traditional Chinese medicine, this might be due to the regulation of “Yin” and “Yang” imbalance of joints to modify the pathological status in rheumatoid arthritis. This study provided an experimental basis for the application of overall alkaloids of TBL in the treatment of rheumatoid arthritis.展开更多
Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunc...Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.展开更多
This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4＋ helper T lymphocytes ...This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4＋ helper T lymphocytes （Th1/Th2） and human leukocyte antigen （HLA）-DR expression on CD14＋ monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha （TNF-α）, interleukin-6 （IL-6）, interleukin-10 （IL-10） and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein （CRP） were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14＋ monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.展开更多
To determine whether RNA interference （RNAi） could block cytotoxic T-lymphocyte antigen 4 （CTLA4） in human lymphocytes in vitro and promote IFN-T and IL-2 secretions, three small interfering RNAs （siRNAs） were s...To determine whether RNA interference （RNAi） could block cytotoxic T-lymphocyte antigen 4 （CTLA4） in human lymphocytes in vitro and promote IFN-T and IL-2 secretions, three small interfering RNAs （siRNAs） were selected based on target specificity sequences of human CTLA4 and transfected into human lymphocytes of chronic HBV patients. As a result, the expression of human CTLA4 mRNA was efficiently suppressed by all the three siRNAs. Compared with negative control （siRNA-co）, siRNA-1 inhibited the expression of CTLA4 most efficiently and was used in the further study. The expressions of IFN-γ and IL-2 were upregulated and the level of IL-4 was almost unchanged in lymphocytes transfected with siRNA-1 compared with the blank control. These results indicated that siRNA-1 led to IFN-γ and IL-2 secretions, which is a main response of Th1/Th2. In a conclusion, RNAi significantly suppressed the expression of human CTLA4 mRNA in human lymphocytes in vitro, and could induce Th1/Th2 response. It could be a new therapeutic strategy for chronic HBV infection.展开更多
Objective： To study the principle of clearing Fei （肺）, cooling blood, and detoxification as well as nourishing yin and moisening Fei （abbr. as CCD-NM） in regulating the levels of peripheral T-lymphocyte subsets ...Objective： To study the principle of clearing Fei （肺）, cooling blood, and detoxification as well as nourishing yin and moisening Fei （abbr. as CCD-NM） in regulating the levels of peripheral T-lymphocyte subsets Th and Tc cells to explore its mechanism for lowering the incidence of infection in patients with systemic lupus erythematosus iSLE）. Methods： Sixty SLE patients without complicated infection were assigned to the treatment group and the control group, 30 in each group. The control group was treated with Western medicine alone, while the treatment group was treated with the same program of Western medicine, but additionally administered with either Langchuang No.1 （狼疮 Ⅰ号） or 2 （狼疮 Ⅱ 号）, serial concerted Chinese recipes, applied respectively in patients in the active stage or in the resting stage. The total time of treatment for both groups was 1 year. Further, a healthy control group was set up with 20 healthy subjects. The expressions of Thl, Th2, and Tcl and Tc2 cells in peripheral blood were detected and compared with those in the healthy control group. Results： （1） As compared with the healthy control group, ratios of Thl/Th2 and Tcl/Tc2 in SLE patients, whether complicated with infection or not, were significantly lower （P〈0.05 or P〈0.01）. （2） Comparison between patients with complications and those uncomplicated with infection showed that the two ratios and Thl expression were lower and Tc2 was higher in the former than those in the latter （all P〈0.05）. （3） Ratios of Thl/Th2 and Tcl/Tc2 increased after treatment in patients of both the treatment group and the control group （P〈0.05 and P〈0.01）, but the changes in the treatment group were more significant （P〈0.05）. Conclusion： The principle of CCD-NM could regulate the Th and Tc subsets toward equilibrium in SLE patients, which might be one of the mechanisms of action for alleviating complicated infection.展开更多
文摘Objective: This study analyzed the T lymphocytes and Th1/Th2 type cytokine profile shift in the peripheral blood ofpatients with recurrent genital herpes (RGH). Methods: Immunofluorescent staining or cell surface antigenand intracellular cytokines(IL-2、IL-4、IL-12、IFN-r)inperipheral blood from 20 RGH patients and 10 controls wereanalyzed using flow cytometric techniques. Results: RGH patients had signiflcantly lower levels ofCD3^+T cells, CD4^+T cells and CD4^+ T/ CD8^+ T cells ratiocompared to control levels (P<0.001), and IL-2-producing,IFN-r-producing and IL-12-producing T cells were increasedin RGH patients (CD4^+T: P<0.001, CD8^+T: P<0.05respectively), whereas IL-4-producing T cells were increased inRGH patients compared to controls (CD4^+T: P<0.05; CD8^+T:P<0.001 respectively). Conclusions: RGH patients have T lymphocyte subsetvariations and Th1/Th2 cytokine changes. The increase in Th2cells Th1/Th2 imbalance may have important implications forRGH pathogenesis.
文摘Objective： To evaluate whether radiofrequency ablation （RFA） might have an influence on immune status in hepatocellular carcinoma （HCC） patients. Methods： We measured the T lymphocytes, B lymphocyte and NK cells, and determined the population of Thl, Th2, Tcl and Tc2 of peripheral blood samples taken from 26 HCC patients before and after RFA. Results： The proportion of Typel cells （Thl and Tcl） and NK cells were significantly increased after RFA, especially in patients of the following subgroups： male, age〉55 years, pathological grade Ⅰ-Ⅱ tumor, clinical stage Ⅰ-Ⅱ or Child-Pugh A and B. Conclusion： TypeⅠ cells and NK cells in HCC patients were increased in a short period after RFA.
文摘To investigate the effects of overall alkali of a traditional Chinese medicine “Tongbiling” (brucine and strychnine alkaloids in main) on the cytokines expression in Th1 and Th2 cells in the synovial fluid of patients with rheumatism arthritis and their signal pathway, the mononuclear cells in the synovial fluid (SFMC) of patients were isolated by Ficoll-Hypaque gradient centrifugation, and the CD3 + CD69 + and CD3 + HLA-DR antigen were analyzed by flow cytometry in comparison with those of the peripheral blood. The rest of cells were cultured after resuspension with RPMI 1640 culture medium. Phorbol 12,13-dibutyrate (PDB) and ionomycin were added successively into the culture with various concentration of overall alkali Tongbiling (TBL). After 4 h of cultivation, the expression of IFN-γ and IL-4 in CD3 + cells were analyzed by flow cytometry. The influence of overall alkali TBL (100?mg/L) on the intracellular calcium was investigated after Fluo-3/AM labeling and stimulation with PDB and ionomycin at 1, 2, 4 and 10?min, and the influence of TBL on the expression of CD3 + CD69 + cells were determined with stimulation of PDB for 24?h in the whole blood lymphocytes culture. It was found that the percentage of T cells bearing CD69 was significantly up-regulated (77%), while that of T cells bearing HLA-DR was 44% in the synovial mononucleated cells. After PDB and ionomycin stimulation, the expression of IFN-γ in CD3 + cells were up-regulated, but there was no change on the expression of IL-4 in CD3 + cells, indicating that ratio of Th1/Th2 was significantly increased and Th cells differentiate to Th1 cells in mainly. Four concentrations of overall alkaloid of TBL (200?mg/L, 100?mg/L, 50?mg/L, 25?mg/L) could down-regulated the expression of IFN-γ in CD3 + cells and the Th1/Th2 ratio obviously, but all the concentrations of the overall alkaloids had no effect on the expression of IL-4 in CD3 + cells. 100?mg/L concentration of the overall alkaloid did not down-regulate the intracellular calcium level. Each concentration of the overall alkaloid could down-regulated the expression CD69 obviously on the PDB-activated mouse T cells. It concluded from the above observations that the overall alkaloid of TBL could relieve the inflammatory and immune damages by suppressing the expression of Th1 type cytokines and Th1 cell differentiation, regulating the imbalance of Th1/Th2 cells and inhibiting the early activation of the T lymphocytes bearing CD69. There was no remarkable influence on the intracellular calcium signaling transduction pathway. The inhibitory effected on T cells to express IFN-γ might be due to the suppression of PKC-MAPK signaling pathway. From the standpoint of traditional Chinese medicine, this might be due to the regulation of “Yin” and “Yang” imbalance of joints to modify the pathological status in rheumatoid arthritis. This study provided an experimental basis for the application of overall alkaloids of TBL in the treatment of rheumatoid arthritis.
基金This study was supported by the National Natural Science Foundation of China(No.30672178)National Basic Research Program of China(No.2005CB522602)the National Natural Science Outstanding Youth Foundation of China(No.30125020).
文摘Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.
基金supported by a grant from the National Natural Sciences Foundation of China(No.30700869)
文摘This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4＋ helper T lymphocytes （Th1/Th2） and human leukocyte antigen （HLA）-DR expression on CD14＋ monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha （TNF-α）, interleukin-6 （IL-6）, interleukin-10 （IL-10） and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein （CRP） were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14＋ monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.
文摘To determine whether RNA interference （RNAi） could block cytotoxic T-lymphocyte antigen 4 （CTLA4） in human lymphocytes in vitro and promote IFN-T and IL-2 secretions, three small interfering RNAs （siRNAs） were selected based on target specificity sequences of human CTLA4 and transfected into human lymphocytes of chronic HBV patients. As a result, the expression of human CTLA4 mRNA was efficiently suppressed by all the three siRNAs. Compared with negative control （siRNA-co）, siRNA-1 inhibited the expression of CTLA4 most efficiently and was used in the further study. The expressions of IFN-γ and IL-2 were upregulated and the level of IL-4 was almost unchanged in lymphocytes transfected with siRNA-1 compared with the blank control. These results indicated that siRNA-1 led to IFN-γ and IL-2 secretions, which is a main response of Th1/Th2. In a conclusion, RNAi significantly suppressed the expression of human CTLA4 mRNA in human lymphocytes in vitro, and could induce Th1/Th2 response. It could be a new therapeutic strategy for chronic HBV infection.
基金the Funds of Hangzhou Municipal Medical Key Subjects(No.2005633Q14)
文摘Objective： To study the principle of clearing Fei （肺）, cooling blood, and detoxification as well as nourishing yin and moisening Fei （abbr. as CCD-NM） in regulating the levels of peripheral T-lymphocyte subsets Th and Tc cells to explore its mechanism for lowering the incidence of infection in patients with systemic lupus erythematosus iSLE）. Methods： Sixty SLE patients without complicated infection were assigned to the treatment group and the control group, 30 in each group. The control group was treated with Western medicine alone, while the treatment group was treated with the same program of Western medicine, but additionally administered with either Langchuang No.1 （狼疮 Ⅰ号） or 2 （狼疮 Ⅱ 号）, serial concerted Chinese recipes, applied respectively in patients in the active stage or in the resting stage. The total time of treatment for both groups was 1 year. Further, a healthy control group was set up with 20 healthy subjects. The expressions of Thl, Th2, and Tcl and Tc2 cells in peripheral blood were detected and compared with those in the healthy control group. Results： （1） As compared with the healthy control group, ratios of Thl/Th2 and Tcl/Tc2 in SLE patients, whether complicated with infection or not, were significantly lower （P〈0.05 or P〈0.01）. （2） Comparison between patients with complications and those uncomplicated with infection showed that the two ratios and Thl expression were lower and Tc2 was higher in the former than those in the latter （all P〈0.05）. （3） Ratios of Thl/Th2 and Tcl/Tc2 increased after treatment in patients of both the treatment group and the control group （P〈0.05 and P〈0.01）, but the changes in the treatment group were more significant （P〈0.05）. Conclusion： The principle of CCD-NM could regulate the Th and Tc subsets toward equilibrium in SLE patients, which might be one of the mechanisms of action for alleviating complicated infection.
