AIM To examine whether nuclear factor kappa B(NF-κB) activity regulates LIN28 B expression and their roles in leukemia stem cell(LSC)-like properties. METHODS We used pharmacological inhibitor and cell viability assa...AIM To examine whether nuclear factor kappa B(NF-κB) activity regulates LIN28 B expression and their roles in leukemia stem cell(LSC)-like properties. METHODS We used pharmacological inhibitor and cell viability assays to examine the relation between NF-κB and LIN28 B. Western blot and q RT-PCR was employed to determine their protein and m RNA levels. Luciferase reporter was constructed and applied to explore the transcriptional regulation of LIN28 B. We manipulated LIN28 B level in acute myeloid leukemia(AML) cells and investigated LSC-like properties with colony forming and serial replating assays. RESULTS This study revealed the relationship between NF-κB and LIN28 B in AML cells through drug inhibition and overexpression experiments. Notably,inhibition of NF-κB by pharmacological inhibitors reduced LIN28 B expression and decreased cell proliferation. We demonstrated that NF-κB binds to the-819 to-811 region of LIN28 B promoter,and transcriptionally regulates LIN28 B expression. LIN28 B protein was significantly elevated in NFκB1 transfected cells compared to vector control. Importantly,ectopic expression of LIN28 B partially rescued the self-renewal capacity impaired by pharmacological inhibition of NF-κB activity. CONCLUSION These results uncover a regulatory signaling,NF-κB/LIN28 B,which plays a pivotal role in leukemia stem cell-like properties and it could serve as a promising intervening target for effective treatment of AML disease.展开更多
Mammalian T-cell responses require synergism between the first signal and co-stimulatory signal.However,whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown.In the present ...Mammalian T-cell responses require synergism between the first signal and co-stimulatory signal.However,whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown.In the present study,we discovered that the Nile tilapia(Oreochromis niloticus)encodes key components of the LAT signalosome,namely,LAT,ITK,GRB2,VAV1,SLP-76,GADS,and PLC-γ1.These components are evolutionarily conserved,and CD3εmAb-induced T-cell activation markedly increased their expression.Additionally,at least ITK,GRB2,and VAV1 were found to interact with LAT for signalosome formation.Downstream of the first signal,the NF-κB,MAPK/ERK,and PI3K-AKT pathways were activated upon CD3εmAb stimulation.Furthermore,treatment of lymphocytes with CD28 mAbs triggered the AKT-mTORC1 pathway downstream of the co-stimulatory signal.Combined CD3εand CD28 mAb stimulation enhanced ERK1/2 and S6 phosphorylation and elevated NFAT1,c-Fos,IL-2,CD122,and CD44 expression,thereby signifying T-cell activation.Moreover,rather than relying on the first or co-stimulatory signal alone,both signals were required for T-cell proliferation.Full T-cell activation was accompanied by marked apoptosis and cytotoxic responses.These findings suggest that tilapia relies on dual signaling to maintain an optimal T-cell response,providing a novel perspective for understanding the evolution of the adaptive immune system.展开更多
Generally, TGF-βs are held to down- regulate the growth of immune cells and to inhibit the development of certain differentiated functions, such as the induction of LAK activity by IL-2. In the present study, the eff...Generally, TGF-βs are held to down- regulate the growth of immune cells and to inhibit the development of certain differentiated functions, such as the induction of LAK activity by IL-2. In the present study, the effects of TGF-β1 on the proliferation and cytotoxicity of human PBMC activated by anti-CD3 or and-CD3 plus anti-CD28 was investigated. The results demonstrated that TGF- β1 clearly inhibits the induction of cytotoxic ability in human PBMC stimulated via CD3 or CD3 and CD28 ( P<0. 01) , without significantly altering the proliferative response to these stimuli, at the tested doses of TGF-β1. Co-stimulation with IL-2 was hardly altered, suggesting that TGF-β1 action is affected by the nature of the costimulatory signals.展开更多
Summary： To investigate the expression and significance of CD28 and CTLA4 on T cells in bone marrow of aplastic anemia （AA） mice, in vitro bone marrow mononuclear cells （BMMNCs） were activated through being incub...Summary： To investigate the expression and significance of CD28 and CTLA4 on T cells in bone marrow of aplastic anemia （AA） mice, in vitro bone marrow mononuclear cells （BMMNCs） were activated through being incubated with PHA （15 μg/mL）. The expression of CD28 and CTLA4 on T cells incubated with or without PHA was detected by two-color flow cytometry. The expression of CD28 and CTLA4 was significantly increased after PHA stimulation. In the AA mice. the expression of CD28 with or without PHA stimulation was both higher than that in the normal mice （both P〈0.01）, but the expression of CTLA4 with or without PHA stimulation showed no significant difference in comparison to that in the normal mice （both P〉0.05）. In the AA mice, there were more activation and activated potential of T cells than the normal, and the abnormal expression of CD28 and CTLA4 may participate in immunological disorder mediated by T cells.展开更多
Objective:To study the changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke and their correlation with neuronal damage markers, inflammatory cytokines and plaque stability indicators.M...Objective:To study the changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke and their correlation with neuronal damage markers, inflammatory cytokines and plaque stability indicators.Methods: The patients who were diagnosed with acute ischemic stroke in our hospital between June 2014 and December 2017 were selected as the stroke group of the research, and healthy volunteers who received physical examination during the same period were selected as the control group. Peripheral blood was collected to determine the contents of CD4+CD25+ and CD4+CD28-T cells, and serum was collected to determine the contents of neuron damage markers, inflammatory cytokines and plaque stability indicators.Results: Peripheral blood CD4+CD25+ cell content as well as serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents of stroke group was lower than those of control group whereas peripheral blood CD4+CD28-T cell content as well as serum NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents was higher than those of control group;peripheral blood CD4+CD25+ cell content of stroke group was positively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and negatively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents;peripheral blood CD4+CD28-T cell content was negatively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and positively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents.Conclusion: The changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke can aggravate the neuron damage and promote the inflammatory response activation and plaque stability decline.展开更多
Previous research found that cerebral dopamine neurotrophic factor(CDNF)has a protective effect on brain dopaminergic neurons,and CDNF is regarded as a promising therapeutic agent for neurodegenerative diseases.Howeve...Previous research found that cerebral dopamine neurotrophic factor(CDNF)has a protective effect on brain dopaminergic neurons,and CDNF is regarded as a promising therapeutic agent for neurodegenerative diseases.However,the effects of CDNF on the proliferation,differentiation,and apoptosis of neural stem cells(NSCs),which are very sensitive to hypoxic environments,remain unknown.In this study,NSCs were extracted from the hippocampi of fetal rats and cultured with different concentrations of CDNF.The results showed that 200 nM CDNF was the optimal concentration for significantly increasing the viability of NSCs under non-hypoxic environmental conditions.Then,the cells were cultured with 200 nM CDNF under the hypoxic conditions of 90%N_2,5%CO_2,and 5%air for 6 hours.The results showed that CDNF significantly improved the viability of hypoxic NSCs and reduced apoptosis among hypoxic NSCs.The detection of markers showed that CDNF increased the differentiation of hypoxic NSCs into neurons and astrocytes.CDNF also reduced the expression level of Lin28 protein and increased the expression of Let-7 mRNA in NSCs,under hypoxic conditions.In conclusion,we determined that CDNF was able to reverse the adverse proliferation,differentiation,and apoptosis effects that normally affect NSCs in a hypoxic environment.Furthermore,the Lin28/Let-7 pathway may be involved in this regulated function of CDNF.The present study was approved by the Laboratory Animal Centre of Southeast University,China(approval No.20180924006)on September 24,2018.展开更多
Adipose mesenchymal stem cells(ADSCs)have protective effects against glutamate-induced excitotoxicity,but ADSCs are limited in use for treatment of optic nerve injury.Studies have shown that the extracellular vesicles...Adipose mesenchymal stem cells(ADSCs)have protective effects against glutamate-induced excitotoxicity,but ADSCs are limited in use for treatment of optic nerve injury.Studies have shown that the extracellular vesicles(EVs)secreted by ADSCs(ADSC-EVs)not only have the function of ADSCs,but also have unique advantages including non-immunogenicity,low probability of abnormal growth,and easy access to target cells.In the present study,we showed that intravitreal injection of ADSC-EVs substantially reduced glutamate-induced damage to retinal morphology and electroretinography.In addition,R28 cell pretreatment with ADSC-EVs before injury inhibited glutamate-induced overload of intracellular calcium,downregulation ofα-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor(AMPAR)subunit GluA2,and phosphorylation of GluA2 and protein kinase C alpha in vitro.A protein kinase C alpha agonist,12-O-tetradecanoylphorbol 13-acetate,inhibited the neuroprotective effects of ADSC-EVs on glutamate-induced R28 cells.These findings suggest that ADSCEVs ameliorate glutamate-induced excitotoxicity in the retina through inhibiting protein kinase C alpha activation.展开更多
基金Supported by the Singapore National Research Foundation and the Ministry of Education under the Research Center of Excellence Program to WJ Chng and NMRC Clinician-Scientist IRG Grant CNIG11nov38(Zhou J)supported by NMRC Clinician Scientist Investigator awardpartially supported by the RNA Biology Center at CSI Singapore,NUS,from funding by the Singapore Ministry of Education’s Tier 3 Grants,No.MOE2014-T3-1-006
文摘AIM To examine whether nuclear factor kappa B(NF-κB) activity regulates LIN28 B expression and their roles in leukemia stem cell(LSC)-like properties. METHODS We used pharmacological inhibitor and cell viability assays to examine the relation between NF-κB and LIN28 B. Western blot and q RT-PCR was employed to determine their protein and m RNA levels. Luciferase reporter was constructed and applied to explore the transcriptional regulation of LIN28 B. We manipulated LIN28 B level in acute myeloid leukemia(AML) cells and investigated LSC-like properties with colony forming and serial replating assays. RESULTS This study revealed the relationship between NF-κB and LIN28 B in AML cells through drug inhibition and overexpression experiments. Notably,inhibition of NF-κB by pharmacological inhibitors reduced LIN28 B expression and decreased cell proliferation. We demonstrated that NF-κB binds to the-819 to-811 region of LIN28 B promoter,and transcriptionally regulates LIN28 B expression. LIN28 B protein was significantly elevated in NFκB1 transfected cells compared to vector control. Importantly,ectopic expression of LIN28 B partially rescued the self-renewal capacity impaired by pharmacological inhibition of NF-κB activity. CONCLUSION These results uncover a regulatory signaling,NF-κB/LIN28 B,which plays a pivotal role in leukemia stem cell-like properties and it could serve as a promising intervening target for effective treatment of AML disease.
基金supported by the National Key Research and Development Program(2022YFD2400804)National Natural Science Foundation of China(32022086,31972822)Natural Science Foundation of Shanghai(20ZR1417500)。
文摘Mammalian T-cell responses require synergism between the first signal and co-stimulatory signal.However,whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown.In the present study,we discovered that the Nile tilapia(Oreochromis niloticus)encodes key components of the LAT signalosome,namely,LAT,ITK,GRB2,VAV1,SLP-76,GADS,and PLC-γ1.These components are evolutionarily conserved,and CD3εmAb-induced T-cell activation markedly increased their expression.Additionally,at least ITK,GRB2,and VAV1 were found to interact with LAT for signalosome formation.Downstream of the first signal,the NF-κB,MAPK/ERK,and PI3K-AKT pathways were activated upon CD3εmAb stimulation.Furthermore,treatment of lymphocytes with CD28 mAbs triggered the AKT-mTORC1 pathway downstream of the co-stimulatory signal.Combined CD3εand CD28 mAb stimulation enhanced ERK1/2 and S6 phosphorylation and elevated NFAT1,c-Fos,IL-2,CD122,and CD44 expression,thereby signifying T-cell activation.Moreover,rather than relying on the first or co-stimulatory signal alone,both signals were required for T-cell proliferation.Full T-cell activation was accompanied by marked apoptosis and cytotoxic responses.These findings suggest that tilapia relies on dual signaling to maintain an optimal T-cell response,providing a novel perspective for understanding the evolution of the adaptive immune system.
文摘Generally, TGF-βs are held to down- regulate the growth of immune cells and to inhibit the development of certain differentiated functions, such as the induction of LAK activity by IL-2. In the present study, the effects of TGF-β1 on the proliferation and cytotoxicity of human PBMC activated by anti-CD3 or and-CD3 plus anti-CD28 was investigated. The results demonstrated that TGF- β1 clearly inhibits the induction of cytotoxic ability in human PBMC stimulated via CD3 or CD3 and CD28 ( P<0. 01) , without significantly altering the proliferative response to these stimuli, at the tested doses of TGF-β1. Co-stimulation with IL-2 was hardly altered, suggesting that TGF-β1 action is affected by the nature of the costimulatory signals.
文摘Summary： To investigate the expression and significance of CD28 and CTLA4 on T cells in bone marrow of aplastic anemia （AA） mice, in vitro bone marrow mononuclear cells （BMMNCs） were activated through being incubated with PHA （15 μg/mL）. The expression of CD28 and CTLA4 on T cells incubated with or without PHA was detected by two-color flow cytometry. The expression of CD28 and CTLA4 was significantly increased after PHA stimulation. In the AA mice. the expression of CD28 with or without PHA stimulation was both higher than that in the normal mice （both P〈0.01）, but the expression of CTLA4 with or without PHA stimulation showed no significant difference in comparison to that in the normal mice （both P〉0.05）. In the AA mice, there were more activation and activated potential of T cells than the normal, and the abnormal expression of CD28 and CTLA4 may participate in immunological disorder mediated by T cells.
文摘Objective:To study the changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke and their correlation with neuronal damage markers, inflammatory cytokines and plaque stability indicators.Methods: The patients who were diagnosed with acute ischemic stroke in our hospital between June 2014 and December 2017 were selected as the stroke group of the research, and healthy volunteers who received physical examination during the same period were selected as the control group. Peripheral blood was collected to determine the contents of CD4+CD25+ and CD4+CD28-T cells, and serum was collected to determine the contents of neuron damage markers, inflammatory cytokines and plaque stability indicators.Results: Peripheral blood CD4+CD25+ cell content as well as serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents of stroke group was lower than those of control group whereas peripheral blood CD4+CD28-T cell content as well as serum NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents was higher than those of control group;peripheral blood CD4+CD25+ cell content of stroke group was positively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and negatively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents;peripheral blood CD4+CD28-T cell content was negatively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and positively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents.Conclusion: The changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke can aggravate the neuron damage and promote the inflammatory response activation and plaque stability decline.
基金supported by the National Natural Science Foundation of China,Nos.81471780 and 81671819(both to LKC)。
文摘Previous research found that cerebral dopamine neurotrophic factor(CDNF)has a protective effect on brain dopaminergic neurons,and CDNF is regarded as a promising therapeutic agent for neurodegenerative diseases.However,the effects of CDNF on the proliferation,differentiation,and apoptosis of neural stem cells(NSCs),which are very sensitive to hypoxic environments,remain unknown.In this study,NSCs were extracted from the hippocampi of fetal rats and cultured with different concentrations of CDNF.The results showed that 200 nM CDNF was the optimal concentration for significantly increasing the viability of NSCs under non-hypoxic environmental conditions.Then,the cells were cultured with 200 nM CDNF under the hypoxic conditions of 90%N_2,5%CO_2,and 5%air for 6 hours.The results showed that CDNF significantly improved the viability of hypoxic NSCs and reduced apoptosis among hypoxic NSCs.The detection of markers showed that CDNF increased the differentiation of hypoxic NSCs into neurons and astrocytes.CDNF also reduced the expression level of Lin28 protein and increased the expression of Let-7 mRNA in NSCs,under hypoxic conditions.In conclusion,we determined that CDNF was able to reverse the adverse proliferation,differentiation,and apoptosis effects that normally affect NSCs in a hypoxic environment.Furthermore,the Lin28/Let-7 pathway may be involved in this regulated function of CDNF.The present study was approved by the Laboratory Animal Centre of Southeast University,China(approval No.20180924006)on September 24,2018.
基金supported by the National Key R&D Program of China,No.2016YFC1201800(to JFH)the Key Research and Development Program of Hunan Province,Nos.2018SK2090(to JFH),2022SK2079(to JFH)+2 种基金the Natural Science Foundation of Hu nan Province,No.2021JJ30891(to DC)the Human Resource Bank Program of Hunan Province,No.2020TP3003(to JFH)the School-Enterprise Joint Program of Central South University,No.2021XQLH092(to TQD)。
文摘Adipose mesenchymal stem cells(ADSCs)have protective effects against glutamate-induced excitotoxicity,but ADSCs are limited in use for treatment of optic nerve injury.Studies have shown that the extracellular vesicles(EVs)secreted by ADSCs(ADSC-EVs)not only have the function of ADSCs,but also have unique advantages including non-immunogenicity,low probability of abnormal growth,and easy access to target cells.In the present study,we showed that intravitreal injection of ADSC-EVs substantially reduced glutamate-induced damage to retinal morphology and electroretinography.In addition,R28 cell pretreatment with ADSC-EVs before injury inhibited glutamate-induced overload of intracellular calcium,downregulation ofα-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor(AMPAR)subunit GluA2,and phosphorylation of GluA2 and protein kinase C alpha in vitro.A protein kinase C alpha agonist,12-O-tetradecanoylphorbol 13-acetate,inhibited the neuroprotective effects of ADSC-EVs on glutamate-induced R28 cells.These findings suggest that ADSCEVs ameliorate glutamate-induced excitotoxicity in the retina through inhibiting protein kinase C alpha activation.
