The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible role...The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.展开更多
Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Inject...Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Injection in treatment of asthma. Methods： 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids （BALF） were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain （HE）. Flow cytometry was used to detect the CD4^＋CD25^＋ Tr ratio in peripheral blood mononuclear cells （PBMCs）. Results： Total cell, the percentage of lymphocytes, neutrophils and eosinophils （Eos） in BALF of Danshen Injection-treated group were lower than that in asthma group （P〈0.05, P〈0.01）. Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4^＋CD25^＋ Tr of asthma group was lower than that of control and Danshen Injection treated group （P〈0.05）. Conclusion： Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4^＋CD25^＋ Tr.展开更多
Objective:To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4^(+)T.Methods:1.PBMC was extracted from peripheral blood mononuclear cells of...Objective:To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4^(+)T.Methods:1.PBMC was extracted from peripheral blood mononuclear cells of children with asthma and normal children in the affiliated Children's Hospital of Soochow University,and RNA was extracted and reverse transcribed.The expression of miR-29b and B7H3mRNA was determined by real-time quantitative polymerase chain reaction(Q-PCR).The family history of asthma and history of allergic diseases were collected.2.THP-1 cells were induced into macrophages,miR-29b interference,miR-29b overexpression and normal control were induced by LV526,LV527 and NC virus infection.After 24 hours of culture,the cells were collected to detect the expression of STAT3 and B7H3 genes and proteins.3.It was verified that STAT3 was the target gene of miR-29b:after inoculating THP-1 cells and culturing with PMA with final concentration of 50ng/ml for 6 hours,the macrophages without PMA were cultured for 24 hours,then the macrophages infected by LV528,LV529 and NC virus were induced to form miR-29b interference,miR29b overexpression and normal control group.Luciferase analysis was performed at 48 hours to verify that STAT3 was the target gene of miR-29b.STAT3-3'UTR luciferase reporter gene plasmids were constructed and divided into three groups:"miR-29b+STAT3-3'UTR","miR-29b+STAT3-mut-3'UTR"and"miR-29b+luciferase empty load".4.Macrophages with different treatments were co-cultured with initial T cells for 3 days.The relative expressions of T-bet,GATA3 and ROR-γt were detected by Q-PCR.Result:1.The incidence of allergic disease in the acute attack group(68%)was higher than that in the other two groups(34.8%,33.3%),and the family history of asthma in the normal group(0%)was much lower than that in the other two groups(52%,60.9%).The difference was statistically significant(P<0.05).2.The expression of B7H3 in PBMC in acute attack group was higher than that in non-acute attack group and normal group.The expression of miR-29b in PBMC in normal group was significantly higher than that in non-acute attack group and acute attack group(P<0.0001).The expression of miR-29b in non-acute attack group was significantly higher than that in acute attack group(P=0.007).3.After silencing the expression of miR-29b,IL-4Rα,IL-4,IL-5,IL-13 and CD206 of macrophages increased significantly,while IFN-γdecreased,suggesting that miR-29b can promote the polarization of macrophages to M2.4.The overexpression of miR-29b,STAT3 and B7H3 gene and protein level in macrophages decreased,while the increase of miR-29b,STAT3 and B7H3 gene and protein expression was inhibited.5.There was a significant positive correlation between the expression of STAT3 and B7H3mRNA in macrophages(r=0.9737,P<0.0001).6.STAT3 is the target gene of miR-29b.7.Co-culture of macrophages with CD4^(+)T cells can promote the differentiation of primary T cells,namely Th 0 cells,into Th2,and the promoting effect of macrophages with downregulation of miR-29b is more obvious.Conclusion:The expression of miR-29b in PBMC of children with asthma is lower than that of normal children,while the expression of B7H3 is higher than that of normal children.It is speculated that miR-29b has a protective effect on children with asthma,while B7H3 aggravates the inflammatory response.Down-regulation of miR-29b,in macrophages can promote macrophages to M2 polarization,increase the expression of B7H3 and STAT3 in macrophages,make Th0 cells differentiate into Th2 cells,and aggravate the inflammatory response in patients with asthma.展开更多
CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance...CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.展开更多
Bronchial asthma is a chronic inflammatory disease of the airways that is characterized by lymphocyte, eosinophil, and mast cell infiltration of the submucosa along with mucous gland hyperplasia and subepithelial fibr...Bronchial asthma is a chronic inflammatory disease of the airways that is characterized by lymphocyte, eosinophil, and mast cell infiltration of the submucosa along with mucous gland hyperplasia and subepithelial fibrosis. The inflammatory response in asthma is tightly associated with airway hyperresponsiveness （AHR） to antigen-specific and nonspecific stimuli. Central to the process of airway inflammation in asthma is the T lymphocyte. Thus, T-cell recruitment and differentiation are critical elements in the evolution of the asthmatic state. Besides adhesion molecules such as selectins and integrins, chemokines are thought to be involved in the multi-step process of extravasation by the triggering of integrins and chemotactic attraction of cell subsets. The diversity of chemokines that potentially attract different cell populations makes them interesting candidates for the regulation of effector cell recruitment. Indeed, chemokines especially are inducible and up-regulated in inflammatory lesions,展开更多
基金This project was supported by a program of Science Project of Hubei Province (No.2003AA301C10).
文摘The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.
基金This This work was supported by a grant from the Science and Technology Foundation of Hubei Province (2003AA301C10)
文摘Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Injection in treatment of asthma. Methods： 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids （BALF） were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain （HE）. Flow cytometry was used to detect the CD4^＋CD25^＋ Tr ratio in peripheral blood mononuclear cells （PBMCs）. Results： Total cell, the percentage of lymphocytes, neutrophils and eosinophils （Eos） in BALF of Danshen Injection-treated group were lower than that in asthma group （P〈0.05, P〈0.01）. Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4^＋CD25^＋ Tr of asthma group was lower than that of control and Danshen Injection treated group （P〈0.05）. Conclusion： Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4^＋CD25^＋ Tr.
基金National Natural Science Foundation of China(No.81570016,81771676,81970027)。
文摘Objective:To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4^(+)T.Methods:1.PBMC was extracted from peripheral blood mononuclear cells of children with asthma and normal children in the affiliated Children's Hospital of Soochow University,and RNA was extracted and reverse transcribed.The expression of miR-29b and B7H3mRNA was determined by real-time quantitative polymerase chain reaction(Q-PCR).The family history of asthma and history of allergic diseases were collected.2.THP-1 cells were induced into macrophages,miR-29b interference,miR-29b overexpression and normal control were induced by LV526,LV527 and NC virus infection.After 24 hours of culture,the cells were collected to detect the expression of STAT3 and B7H3 genes and proteins.3.It was verified that STAT3 was the target gene of miR-29b:after inoculating THP-1 cells and culturing with PMA with final concentration of 50ng/ml for 6 hours,the macrophages without PMA were cultured for 24 hours,then the macrophages infected by LV528,LV529 and NC virus were induced to form miR-29b interference,miR29b overexpression and normal control group.Luciferase analysis was performed at 48 hours to verify that STAT3 was the target gene of miR-29b.STAT3-3'UTR luciferase reporter gene plasmids were constructed and divided into three groups:"miR-29b+STAT3-3'UTR","miR-29b+STAT3-mut-3'UTR"and"miR-29b+luciferase empty load".4.Macrophages with different treatments were co-cultured with initial T cells for 3 days.The relative expressions of T-bet,GATA3 and ROR-γt were detected by Q-PCR.Result:1.The incidence of allergic disease in the acute attack group(68%)was higher than that in the other two groups(34.8%,33.3%),and the family history of asthma in the normal group(0%)was much lower than that in the other two groups(52%,60.9%).The difference was statistically significant(P<0.05).2.The expression of B7H3 in PBMC in acute attack group was higher than that in non-acute attack group and normal group.The expression of miR-29b in PBMC in normal group was significantly higher than that in non-acute attack group and acute attack group(P<0.0001).The expression of miR-29b in non-acute attack group was significantly higher than that in acute attack group(P=0.007).3.After silencing the expression of miR-29b,IL-4Rα,IL-4,IL-5,IL-13 and CD206 of macrophages increased significantly,while IFN-γdecreased,suggesting that miR-29b can promote the polarization of macrophages to M2.4.The overexpression of miR-29b,STAT3 and B7H3 gene and protein level in macrophages decreased,while the increase of miR-29b,STAT3 and B7H3 gene and protein expression was inhibited.5.There was a significant positive correlation between the expression of STAT3 and B7H3mRNA in macrophages(r=0.9737,P<0.0001).6.STAT3 is the target gene of miR-29b.7.Co-culture of macrophages with CD4^(+)T cells can promote the differentiation of primary T cells,namely Th 0 cells,into Th2,and the promoting effect of macrophages with downregulation of miR-29b is more obvious.Conclusion:The expression of miR-29b in PBMC of children with asthma is lower than that of normal children,while the expression of B7H3 is higher than that of normal children.It is speculated that miR-29b has a protective effect on children with asthma,while B7H3 aggravates the inflammatory response.Down-regulation of miR-29b,in macrophages can promote macrophages to M2 polarization,increase the expression of B7H3 and STAT3 in macrophages,make Th0 cells differentiate into Th2 cells,and aggravate the inflammatory response in patients with asthma.
基金the National Research Foundation of Korea(NRF-2021M3A9D3026428)the Ministry of Science and ICT of Korea.J.Jeong is a recipient of funding from the Global Ph.D.Fellowship Program(NRF-2019H1A2A1076865)of the National Research Foundation of Korea.
文摘CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.
基金This study was supported in part by the National Natural Science Foundation of China (No. 30460051), and in part by the Program NCET-04-0835 for New Century Excellent Talents in Chinese Universities.
文摘Bronchial asthma is a chronic inflammatory disease of the airways that is characterized by lymphocyte, eosinophil, and mast cell infiltration of the submucosa along with mucous gland hyperplasia and subepithelial fibrosis. The inflammatory response in asthma is tightly associated with airway hyperresponsiveness （AHR） to antigen-specific and nonspecific stimuli. Central to the process of airway inflammation in asthma is the T lymphocyte. Thus, T-cell recruitment and differentiation are critical elements in the evolution of the asthmatic state. Besides adhesion molecules such as selectins and integrins, chemokines are thought to be involved in the multi-step process of extravasation by the triggering of integrins and chemotactic attraction of cell subsets. The diversity of chemokines that potentially attract different cell populations makes them interesting candidates for the regulation of effector cell recruitment. Indeed, chemokines especially are inducible and up-regulated in inflammatory lesions,
