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AIM: To investigate the regulatory function of bone marrow-derived mesenchymal stem cells (MSCs) on T helper 17 cells (Th17) and regulatory T cells (Treg) in peripheral blood of severe asthmatic children . METHODS:MSCs were isolated , cultured and identified in vitro.MSCs digested with mitomycin were cocultured with T lymphocytes (TLC) at different ratios (1∶1, 1∶2, 1∶10 and 1∶20) from severe asthmatic children for 72 h.The prolifera-tion of TLC was measured by CCK-8 method.In the coculture system of the 1∶2 ratio and the single TLC system , the super-natant levels of interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) were measured by ELISA.The mRNA expression of retinoic acid-related orphan nuclear receptor C (RORC) and forkhead box protein 3 (Foxp3) in TLC was de-tected by qRT-PCR.RESULTS:After cocultured with MSCs , the proliferation of TLC decreased significantly in a dose-dependent manner (P0.05).CONCLUSION: MSCs suppresses Th17 polarization of naive peripheral blood CD 4 +T cells and matures Th17 cells secreting IL-17, which may ef-fectively revise Th17/Treg imbalance of asthma .
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[Objective] This study was designed to determine Th1, Th2 cell numbers and investigate T-bet mRNA, GATA-3 mRNA expression of spleen MNC in a mufine asthmatic model which intended to understand effect of airway T-bet plasmid gene transfer on Th differentiation. [ Methods] A mouse asthmatic model was established by sensitization with ovalbumin (OVA). Thirty-two C57BL/6 mice were divided into four groups (8 mice in each group): the normal control group (group A ), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), the pcDNA3-T-bet group (group D). All animals were sensitized and challenged with OVA, except group A normal saline was applied. The group C was intranasally administered 50 μg pcDNA3 plasmid at 24 h before intranasal challenges, and the 50 μg pcDNA3-T-bet plasmid for the mice of group D. We investigated Th1 and Th2 cell numbers by FACS and T-bet, GATA-3mRNA expression of spleen mononuclear cells (MNC) by semi-quantitative PCR in the four groups. [Result] Th1 percent in spleen MNC of pcDNA3-T-bet treated mice was significantly increased ([2.29±1.551% vs. [1.93±1.141%, P<0.05), while Th2 percent was significantly decreased ([0.93±0.64]% vs. [1.63±0.59]%), compared with that of the asthmatic control group mice by FACS. Spleen MNC was detected a high level of T-bet mRNA expression (0.53±0.027 vs. 0.28±0.035, P<0.05) and a low level of GATA-3 mRNA expression (0.24±0.022 vs. 0.58±0.038, P<0.05) after pcDNA3-T-bet treatment by RT-PCR. There was no significant change between the pcDNA3 plasmid group and the asthmatic model group. [Conclusion] The intranasal transfer of pcDNA3-T-bet plasmid was effective in modulating the imbalance of Th1/Th2 in mice asthma model, which provides a novel therapeutic strategy for transferring transcriptional factor in allergic asthma.
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[Objective] To investigate the changes of CD4~+ CD25~+ regulatory T cells (Tr) in peripheral blood and their relation with their body mass index (BMI) of children with acute attack asthma. [Methods] Peripheral blood was obtained from 70 children with acute attack asthma, 30 remission children, and 50 normal control children. Then 70 children with acute attack asthma, were divided by normal weight group (40 cases) and overweight group (30 cases). The levels of CD4~+CD25~+Tr of the patients were tested by flow cytometry (FCM), and their BMI were calculated. [ Results] The levels of CD4~+ CD25~+ Tr [(6.17± 1.72)%] in acute attack group were lower than that in remission group [(7.56±1.48)%] or that in the control group [(7.13± 1.48)%] (P<0.05), but no difference between that in the remission and that in the control (P>0.05). The CD4~+CD25~+Tr of asthmatic children with normal weight [(6.34±1.71)%] was higher than that of asthmatic children with overweight [(4.74±1.20)%] (P<0.05). There was a remarkably negative correlation between the level of CD4~+ CD25~+ Tr of asthmatic children [(6.17±1.72)%] and the BMI (16.00±2.14) (r_p=-0.814, P<0.05). [Conclusion] The levels of CD4~+ CD25~+Tr are remarkable decrease in attack asthmatic children, and more decrease in overweight patients. There is remarkably negative correlation between the levels of CD4~+CD25~+ Tr in peripheral blood of attack asthmatic children and their BMI.
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AIM: To investigate the effect of glucocorticoid inhalation on the levels of CD4~+CD25~+ regulatory T cells in peripheral blood of asthmatic children. METHODS: Glucocorticoid inhalator was inhaled by 70 children with attack asthma. The levels of CD4~+CD25~+ Tr in peripheral blood of asthmatic children were tested by flow cytometry (FCM). RESULTS: The CD4~+CD25~+ Tr levels in peripheral blood of asthmatic children were (5.62% ± 1.29% ) and (7.05% ± 1.61%) before and after of regulated glucocorticoid inhalation, respectively (P<0.01). The Tr levels were (7.56% ± 1.88% ) , (7.09% ± 1.23% ) and (6.11% ± 1.96% ) in the complete control group, part control group and poor control group, respectively ( P < 0.05 ). The Tr level in formal treatment group (7.05% ±1.61%) was higher than that in irregular treatment group ( 5.91 % ± 1.76% ), P < 0.01.CONCLUSION: The level of CD4~+CD25~+ Tr is remarkable increased by regulated glucocorticoid inhalation, and the level of Tr can reflect the effects of glucocorticoid inhalation.
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AIM: To investigate the effect of T-bet plasmid gene transfer to airway on allergen induced airway inflammation in a murine asthmatic model. METHODS: A mouse asthma model was established by sensitization with ovalbumin (OVA). Forty C57BL/6 mice were divided into 4 groups (10 mice in each group): the normal control group (group A), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), and the pcDNA3-T-bet group (group D). The animals in group B, C and D were sensitized and challenged with OVA. The animals in group A were applied with normal saline. pcDNA3 plasmid at dose of 50 μg was intranasally administered at 24 h before intranasal challenges to the mice in group C, and the 50 μg pcDNA3-T-bet plasmid for the mice in group D. Bronchial alveolar lavage fluid (BALF) was collected and lung tissues were resected at 48 h after OVA challenge for later assay. RESULTS: After administration with pcDNA3-T-bet plasmid, high level of T-bet expression at 48 h was detected in the lung tissue by Western blotting. In pcDNA3-T-bet treated asthmatic models, histological evaluation revealed the significant suppression of eosinophil peribronchial and perivascular infiltration, and reduction of epithelial damage. The numbers of eosinophils, neutrophils and lymphocytes in BALF from pcDNA3-T-bet treated mice were significantly reduced compared to those in asthmatic control group (P<0.05). The level of IL-4 in BALF was significantly decreased in pcDNA3-T-bet group compared to that in asthmatic control group (P<0.05), while the level of IFN-γ in BALF was significantly increased in pcDNA3-T-bet group. No significant change of inflammation cells and cytokines in pcDNA3 plasmid group and asthmatic control group was observed (P>0.05). CONCLUSION: Intranasal pcDNA3-T-bet plasmid transfer inhibits asthmatic airway inflammation in the murine asthmatic model, suggesting a new therapeutic strategy for allergic asthma.
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In this study,IL-4 and IL-12 levels in plasma and peripheral blood mononuclear cells(PBMC) as well as serum IgE level were prospectively assessed with double-antibody sandwich ELISA technique in children with asthma during the attack group (30 cases)and the interval group(12 cases).The results observed revealed that serum IgE level and IL-4 level in plasma and PBMC after PHA and LPS provocation during both of the attack stage and the interval stage were found to be evidently higher than those of the normal control group (P＜0.01).Otherwise,IL-12 level during both stages was much lower than that of the normal control(P＜0.01).In other hand,there were found to have a significant difference in all these 3 data between attack and interval stages.Thus, the conclusion indicates that there might be an imbalance of IL-4,IL-12 and IgE level in children with asthma during both of the attack and the interval stages.
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AIM and METHODS: To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group. RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13. CONCLUSION: Measles vaccine could down-regulate IL-13 level, therefore decrease total IgE synthesis, but not affect IL-12 level in asthmatic children.