IMPAIRED FUNCTION OF MESENCHYMAL STEM CELLS FROM IMMUNE THROMBOCYTOPENIA PATIENTS IN INDUCING REGULATORY DENDRITIC CELL DIFFERENTIATION VIA NOTCH2/JAGGED-1 SIGNALLING PATHWAY
(Abstract release date: 05/19/16)
EHA Library. Su Y. 06/12/16; 135318; S824
Prof. Yan Su
Contributions
Contributions
Abstract
Abstract: S824
Type: Oral Presentation
Presentation during EHA21: On Sunday, June 12, 2016 from 08:15 - 08:30
Location: Room H4
Background
Immune thrombocytopenia(ITP) is characterized by platelet destruction and megakaryocyte dysfunction due to the breakdown of self-tolerance. Mesenchymal stem cells(MSCs) play a pivotal role in immune tolerance. Bone marrow MSCs(BMMSCs) residing in bone marrow stroma (BMS)have been shown to induce regulatory dendritic cell(regDC) differentiation from CD34+ haematopoietic progenitor cells (HPCs). MSCs from ITP patients (ITP-MSCs) exhibit increased senescence and apoptosis, but whether ITP-MSCs have an impaired ability to instruct regDCs remains unknown. Notch is a major factor mediating the interaction between HPCs and BMS.BMS has been reported to prevent the terminal differentiation of DCs through the Notch ligand Jagged-1. Moreover, Jagged-1 has been shown to be involved in the immunosuppressive effects of MSCs on T cells and DCs. It has shown that Notch signalling is important for the modulation of MSCs.
Aims
To investigate whether ITP-MSCs displayed impaired function in inducing regulatory dendritic cell differentiation via notch2/jagged-1 signalling pathway.
Methods
CD34+HPCs were differentiated in the absence or presence of ITP-MSCs or control-MSCs, and then immature DCs were harvested and induced to mature with LPS in the absence of MSCs. MGG staining, flow cytometry and ELISA were performed to characterize the DCs. Mature DCs were then cocultured with CD4+T cells, and proliferation, differentiation, and the anergic and regulatory features of T cells were assessed by flow cytometry. The differential expression of notch receptors, notch intracellular domain(NICD),hes-1, and Jagged-1, was assessed by RT-PCR, flow cytometry and western blot analysis.NICD2-expressing lentivirus,Notch2-targeting shRNA lentivirus, exogenous Jagged-1, and anti-Jagged-1 Ab were used to analyse the underlying mechanism(s).
Results
ITP-MSCs showed impaired function in inhibiting morphological and phenotypic development, proliferation, interleukin-12(IL-12) secretion, and in enhancing endocytosis and interleukin-10(IL-10) production of DCs. DCs that were differentiated in the presence of ITP-MSCs(ITP-MSC-DCs) exhibited a dramatically impaired ability to inhibit allogeneic CD4+ T cell proliferation and to change Th2 polarization. The ITP-MSC-DCs had a deficient capacity to induce anergic and regulatory T cells.Jagged-1 expression and Notch2 signalling pathway activation were down-regulated in ITP-MSCs. The application of exogenous Jagged-1 and anti-Jagged-1 Ab revealed an involvement of Jagged-1 in the development of regDCs. And NICD2 overexpression of ITP-MSCs displayed elevated Jagged-1 expression and an enhanced ability to induce regDCs, which was suppressed after treatment with anti-Jagged-1 Ab. The attenuated function of control-MSCs following Notch2 knockdown was reversed by exogenous Jagged-1. Thus, the Notch2 signalling pathway may modulate regDC differentiation in a Jagged-1-dependent manner. Following pre-treatment with aspirin, both ITP-MSCs and control-MSCs exhibited an up-regulation of the Notch2/Jagged-1 signalling pathway and an enhanced ability to induce regDCs.
Conclusion
ITP-MSCs show defects in the induction of regDC development, which may play a role in the pathogenesis of ITP. The Notch2/Jagged-1 signalling pathway is involved in the impaired function of ITP-MSCs, and agents targeting this pathway may have great potential for the treatment of ITP.
Session topic: Platelet disorders 2
Keyword(s): Dendritic cell, Immune thrombocytopenia (ITP), Mesenchymal stem cell, Notch signaling
Type: Oral Presentation
Presentation during EHA21: On Sunday, June 12, 2016 from 08:15 - 08:30
Location: Room H4
Background
Immune thrombocytopenia(ITP) is characterized by platelet destruction and megakaryocyte dysfunction due to the breakdown of self-tolerance. Mesenchymal stem cells(MSCs) play a pivotal role in immune tolerance. Bone marrow MSCs(BMMSCs) residing in bone marrow stroma (BMS)have been shown to induce regulatory dendritic cell(regDC) differentiation from CD34+ haematopoietic progenitor cells (HPCs). MSCs from ITP patients (ITP-MSCs) exhibit increased senescence and apoptosis, but whether ITP-MSCs have an impaired ability to instruct regDCs remains unknown. Notch is a major factor mediating the interaction between HPCs and BMS.BMS has been reported to prevent the terminal differentiation of DCs through the Notch ligand Jagged-1. Moreover, Jagged-1 has been shown to be involved in the immunosuppressive effects of MSCs on T cells and DCs. It has shown that Notch signalling is important for the modulation of MSCs.
Aims
To investigate whether ITP-MSCs displayed impaired function in inducing regulatory dendritic cell differentiation via notch2/jagged-1 signalling pathway.
Methods
CD34+HPCs were differentiated in the absence or presence of ITP-MSCs or control-MSCs, and then immature DCs were harvested and induced to mature with LPS in the absence of MSCs. MGG staining, flow cytometry and ELISA were performed to characterize the DCs. Mature DCs were then cocultured with CD4+T cells, and proliferation, differentiation, and the anergic and regulatory features of T cells were assessed by flow cytometry. The differential expression of notch receptors, notch intracellular domain(NICD),hes-1, and Jagged-1, was assessed by RT-PCR, flow cytometry and western blot analysis.NICD2-expressing lentivirus,Notch2-targeting shRNA lentivirus, exogenous Jagged-1, and anti-Jagged-1 Ab were used to analyse the underlying mechanism(s).
Results
ITP-MSCs showed impaired function in inhibiting morphological and phenotypic development, proliferation, interleukin-12(IL-12) secretion, and in enhancing endocytosis and interleukin-10(IL-10) production of DCs. DCs that were differentiated in the presence of ITP-MSCs(ITP-MSC-DCs) exhibited a dramatically impaired ability to inhibit allogeneic CD4+ T cell proliferation and to change Th2 polarization. The ITP-MSC-DCs had a deficient capacity to induce anergic and regulatory T cells.Jagged-1 expression and Notch2 signalling pathway activation were down-regulated in ITP-MSCs. The application of exogenous Jagged-1 and anti-Jagged-1 Ab revealed an involvement of Jagged-1 in the development of regDCs. And NICD2 overexpression of ITP-MSCs displayed elevated Jagged-1 expression and an enhanced ability to induce regDCs, which was suppressed after treatment with anti-Jagged-1 Ab. The attenuated function of control-MSCs following Notch2 knockdown was reversed by exogenous Jagged-1. Thus, the Notch2 signalling pathway may modulate regDC differentiation in a Jagged-1-dependent manner. Following pre-treatment with aspirin, both ITP-MSCs and control-MSCs exhibited an up-regulation of the Notch2/Jagged-1 signalling pathway and an enhanced ability to induce regDCs.
Conclusion
ITP-MSCs show defects in the induction of regDC development, which may play a role in the pathogenesis of ITP. The Notch2/Jagged-1 signalling pathway is involved in the impaired function of ITP-MSCs, and agents targeting this pathway may have great potential for the treatment of ITP.
Session topic: Platelet disorders 2
Keyword(s): Dendritic cell, Immune thrombocytopenia (ITP), Mesenchymal stem cell, Notch signaling
Abstract: S824
Type: Oral Presentation
Presentation during EHA21: On Sunday, June 12, 2016 from 08:15 - 08:30
Location: Room H4
Background
Immune thrombocytopenia(ITP) is characterized by platelet destruction and megakaryocyte dysfunction due to the breakdown of self-tolerance. Mesenchymal stem cells(MSCs) play a pivotal role in immune tolerance. Bone marrow MSCs(BMMSCs) residing in bone marrow stroma (BMS)have been shown to induce regulatory dendritic cell(regDC) differentiation from CD34+ haematopoietic progenitor cells (HPCs). MSCs from ITP patients (ITP-MSCs) exhibit increased senescence and apoptosis, but whether ITP-MSCs have an impaired ability to instruct regDCs remains unknown. Notch is a major factor mediating the interaction between HPCs and BMS.BMS has been reported to prevent the terminal differentiation of DCs through the Notch ligand Jagged-1. Moreover, Jagged-1 has been shown to be involved in the immunosuppressive effects of MSCs on T cells and DCs. It has shown that Notch signalling is important for the modulation of MSCs.
Aims
To investigate whether ITP-MSCs displayed impaired function in inducing regulatory dendritic cell differentiation via notch2/jagged-1 signalling pathway.
Methods
CD34+HPCs were differentiated in the absence or presence of ITP-MSCs or control-MSCs, and then immature DCs were harvested and induced to mature with LPS in the absence of MSCs. MGG staining, flow cytometry and ELISA were performed to characterize the DCs. Mature DCs were then cocultured with CD4+T cells, and proliferation, differentiation, and the anergic and regulatory features of T cells were assessed by flow cytometry. The differential expression of notch receptors, notch intracellular domain(NICD),hes-1, and Jagged-1, was assessed by RT-PCR, flow cytometry and western blot analysis.NICD2-expressing lentivirus,Notch2-targeting shRNA lentivirus, exogenous Jagged-1, and anti-Jagged-1 Ab were used to analyse the underlying mechanism(s).
Results
ITP-MSCs showed impaired function in inhibiting morphological and phenotypic development, proliferation, interleukin-12(IL-12) secretion, and in enhancing endocytosis and interleukin-10(IL-10) production of DCs. DCs that were differentiated in the presence of ITP-MSCs(ITP-MSC-DCs) exhibited a dramatically impaired ability to inhibit allogeneic CD4+ T cell proliferation and to change Th2 polarization. The ITP-MSC-DCs had a deficient capacity to induce anergic and regulatory T cells.Jagged-1 expression and Notch2 signalling pathway activation were down-regulated in ITP-MSCs. The application of exogenous Jagged-1 and anti-Jagged-1 Ab revealed an involvement of Jagged-1 in the development of regDCs. And NICD2 overexpression of ITP-MSCs displayed elevated Jagged-1 expression and an enhanced ability to induce regDCs, which was suppressed after treatment with anti-Jagged-1 Ab. The attenuated function of control-MSCs following Notch2 knockdown was reversed by exogenous Jagged-1. Thus, the Notch2 signalling pathway may modulate regDC differentiation in a Jagged-1-dependent manner. Following pre-treatment with aspirin, both ITP-MSCs and control-MSCs exhibited an up-regulation of the Notch2/Jagged-1 signalling pathway and an enhanced ability to induce regDCs.
Conclusion
ITP-MSCs show defects in the induction of regDC development, which may play a role in the pathogenesis of ITP. The Notch2/Jagged-1 signalling pathway is involved in the impaired function of ITP-MSCs, and agents targeting this pathway may have great potential for the treatment of ITP.
Session topic: Platelet disorders 2
Keyword(s): Dendritic cell, Immune thrombocytopenia (ITP), Mesenchymal stem cell, Notch signaling
Type: Oral Presentation
Presentation during EHA21: On Sunday, June 12, 2016 from 08:15 - 08:30
Location: Room H4
Background
Immune thrombocytopenia(ITP) is characterized by platelet destruction and megakaryocyte dysfunction due to the breakdown of self-tolerance. Mesenchymal stem cells(MSCs) play a pivotal role in immune tolerance. Bone marrow MSCs(BMMSCs) residing in bone marrow stroma (BMS)have been shown to induce regulatory dendritic cell(regDC) differentiation from CD34+ haematopoietic progenitor cells (HPCs). MSCs from ITP patients (ITP-MSCs) exhibit increased senescence and apoptosis, but whether ITP-MSCs have an impaired ability to instruct regDCs remains unknown. Notch is a major factor mediating the interaction between HPCs and BMS.BMS has been reported to prevent the terminal differentiation of DCs through the Notch ligand Jagged-1. Moreover, Jagged-1 has been shown to be involved in the immunosuppressive effects of MSCs on T cells and DCs. It has shown that Notch signalling is important for the modulation of MSCs.
Aims
To investigate whether ITP-MSCs displayed impaired function in inducing regulatory dendritic cell differentiation via notch2/jagged-1 signalling pathway.
Methods
CD34+HPCs were differentiated in the absence or presence of ITP-MSCs or control-MSCs, and then immature DCs were harvested and induced to mature with LPS in the absence of MSCs. MGG staining, flow cytometry and ELISA were performed to characterize the DCs. Mature DCs were then cocultured with CD4+T cells, and proliferation, differentiation, and the anergic and regulatory features of T cells were assessed by flow cytometry. The differential expression of notch receptors, notch intracellular domain(NICD),hes-1, and Jagged-1, was assessed by RT-PCR, flow cytometry and western blot analysis.NICD2-expressing lentivirus,Notch2-targeting shRNA lentivirus, exogenous Jagged-1, and anti-Jagged-1 Ab were used to analyse the underlying mechanism(s).
Results
ITP-MSCs showed impaired function in inhibiting morphological and phenotypic development, proliferation, interleukin-12(IL-12) secretion, and in enhancing endocytosis and interleukin-10(IL-10) production of DCs. DCs that were differentiated in the presence of ITP-MSCs(ITP-MSC-DCs) exhibited a dramatically impaired ability to inhibit allogeneic CD4+ T cell proliferation and to change Th2 polarization. The ITP-MSC-DCs had a deficient capacity to induce anergic and regulatory T cells.Jagged-1 expression and Notch2 signalling pathway activation were down-regulated in ITP-MSCs. The application of exogenous Jagged-1 and anti-Jagged-1 Ab revealed an involvement of Jagged-1 in the development of regDCs. And NICD2 overexpression of ITP-MSCs displayed elevated Jagged-1 expression and an enhanced ability to induce regDCs, which was suppressed after treatment with anti-Jagged-1 Ab. The attenuated function of control-MSCs following Notch2 knockdown was reversed by exogenous Jagged-1. Thus, the Notch2 signalling pathway may modulate regDC differentiation in a Jagged-1-dependent manner. Following pre-treatment with aspirin, both ITP-MSCs and control-MSCs exhibited an up-regulation of the Notch2/Jagged-1 signalling pathway and an enhanced ability to induce regDCs.
Conclusion
ITP-MSCs show defects in the induction of regDC development, which may play a role in the pathogenesis of ITP. The Notch2/Jagged-1 signalling pathway is involved in the impaired function of ITP-MSCs, and agents targeting this pathway may have great potential for the treatment of ITP.
Session topic: Platelet disorders 2
Keyword(s): Dendritic cell, Immune thrombocytopenia (ITP), Mesenchymal stem cell, Notch signaling
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