Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated ...Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated by PGE2 and TNF-alpha respectively. The markers of maturation and activation expressed on DCs, such as CD40, CD80, CD83, CD86, MHC-II, and the ability of antigen uptake of DCs were analyzed by flow cytometry. The proliferation of T cells co-cultured with DCs, the signaling pathways of PGE2-EP4-cAMP and TNF-alpha- TRADD-TRAF2-NF-KB in DCs were analyzed. Result Both PGE2 and TNF-alpha up-regulated the expressions of CD40, CD80, CD83, CD86, and MHC-II, decreased the antigen uptake of DCs, and DCs stimulated by PGE2 or TNF-alpha could increase T cell proliferation. CP-25 （ 10 -5, 10 -6 , 10 -7 mol/L ） decreased significantly the ex- pressions of CD40, CD80, CD83, CD86 and MHC- ]I , increased the antigen uptake of DCs, and suppressed T cell proliferation induced by DCs. PGE2 increased the expressions of EP4, NF-KB and down-regulated cAMP level of DCs. TNF-alpha could also up-regulate TNFR1, TRADD, TRAF2, and NF-KB expression of DCs. CP-25 （10^-5, 10^-6, 10^-7 mol/L） decreased the expressions of EP4 and NF-KB, increased cAMP level in DCs stimulated by PGE2. CP-25 （10^-5 10^-6 10^-7 mol/L） also could down-regulate significantly TNFR1 TRADD TRAF2 and NF-KB expression in DCs stimulated by TNF-alpha. Conclusion PGE2 and TNF-alpha could enhance DCs func- tions by mediating PGE2-EP4-cAMP pathway, TNF-alpha-TNFR1-TRADD-TRAF2-NF-KB pathway respectively. CP-25 might inhibit the function of DCs through regulating PGE2-EP4-cAMP and TNF-alpha-TNFR1-TRADD- TRAF2-NF-KB pathways.展开更多
AIM: To study the effect of ginsenoside Rgl and Rhl on the anti - tumor activity of dendritic cells (DC). METHODS: Effect of Rgl and Rhl on the production of IL- 12 p40 protein was detected by ELISA, and the IL- 12 p4...AIM: To study the effect of ginsenoside Rgl and Rhl on the anti - tumor activity of dendritic cells (DC). METHODS: Effect of Rgl and Rhl on the production of IL- 12 p40 protein was detected by ELISA, and the IL- 12 p40 mRNA level of DC was monitored by RT- PCR. Anti - tumor activity of DC- LPAK was detemnined by neutral red staining assay. RESULTS: The results of ELLSA showed that Rgl and Rhl significantly enhanced the production of IL- 12 p40 of DC. Rgl at 1 mg/L and Rhl at 100 mg/L upregulated the IL- 12 p40 mRNA level. Rgl and Rhl enhanced the anti - tumor ability of DC, induced lyrnphokine and PHA activated killer (DC-LPAK) on human papillate tumor cell line. Each dose of Rgl can obviously accelerate the eytotoxity to L929 at the E: T ratio of 5 : 1 (P<0.05,0.01 ), while only Rhl 10 mg/L enhanced the eytotoxity ability of DC- LPAK (P < 0.05). CONCLUSION: Rgl and Rhl enhanced the production of IL-12 p40. This effect may be mediated by the increase in the mRNA level. As a result, Rg1 and Rhl oromote the ability of DC to stimulate the cytotoxitie aetieity of DC - LPAK.展开更多
Dendritic cells （DCs） are bone marrow-derived professional antigen presenting cells （APCs）, they are crucial for initiation of both innate and adaptive immune responses. In this study, chicken bone marrow （chBM）...Dendritic cells （DCs） are bone marrow-derived professional antigen presenting cells （APCs）, they are crucial for initiation of both innate and adaptive immune responses. In this study, chicken bone marrow （chBM） cells were cultured in medium with recombinant chicken granulocyte-macrophage colony stimulating factor （rGM-CSF） and recombinant chicken interleukin-4 （rIL-4） for 7 days, displayed the typical morphology of DCs. These immature chicken bone marrow-derived DCs （chBM-DCs） showed signifcant up-regulation of the putative CD11c and of major histocompatibility complex class II （MHC II）, but CD40 and CD86 co-stimulatory molecules were almost no up-regulated. However, maturation with lipopolysaccharide （LPS）, surface expression of CD40, CD86 was greatly increased. The phagocytosis of chBM-DCs was assessed by neutral red, and the phagocytosis decreased after stimulation. In mixed lymphocyte responses （MLR）, stimulated chBM-DCs were more effective to T-cell stimulators than non-stimulated chBM-DCs. In addition, mRNA expression levels of IL-1β, IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α, CXCLi1 and CXCLi2 were assessed by real-time qPCR （qRT-PCR）, and the results showed cultured chBM-DCs could be matured to a T helper cell type 1 （Th1）-promoting phenotype by LPS stimulation.展开更多
A series of shape-persistent polyphenylene dendritic C_(60)derivatives as the electron transport materials were designed and synthesized via a catalyst-free Diels-Alder[4+2]cycloaddition reaction.These increasing hype...A series of shape-persistent polyphenylene dendritic C_(60)derivatives as the electron transport materials were designed and synthesized via a catalyst-free Diels-Alder[4+2]cycloaddition reaction.These increasing hyperbranched scaffolds could effectively enhance the solubility;notably,both first and second generation dendrimers,C_(60)-G1 and C_(60)-G2,demonstrated more than 5 times higher solubilities than pristine C_(60).Furthermore,both simulated and experimental data proved their promising solution-processabilities as electron-transporting layers(ETLs)for perovskite solar cells.As a result,the planar p-i-n structural perovskite solar cell could achieve a maximum power conversion efficiency of 14.7%with C_(60)-G2.展开更多
Dendritic cells(DCs) are professinal antigen-presenting cells with the ability to initiate primary Tcell responses. While it iswell known that inflammatory stimuli induce the functional maturation of immature DCs, whe...Dendritic cells(DCs) are professinal antigen-presenting cells with the ability to initiate primary Tcell responses. While it iswell known that inflammatory stimuli induce the functional maturation of immature DCs, whether adhesion molecule selectins regulate DCmaturation is poorly understood. Using anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb) that blocks the adhesionof P-, E-, and L-selectin, we demonstrate herein that selectins play important role in stimulating functional maturation of immature DCs.Immature DCs are generated from human cord blood CD34+hematopoietic stem/progenitor cells that were cultured in the presence of stemcell factor, Fms-like tyrosine-kinase-3 ligand, granulocyte-macrophage colony stimulating factor, and transform growth factor-β1. Whenstimulated with tumor necrosis factor-alpha(TNF-α), immature DCs differentiated into mature DCs, producing increased levels of costim-ulatory molecules and interleukin (IL)-12 and obtaining the ability to potently activate naive Tcells. Interestingly, in contrast to matureDCs derived from TNF-α-induced immature DC cultures without PsL-EGFmAb, immature DCs treated with PsL-EGFmAb for7 days werecompletely blocked their maturation, as evidenced by decreased expression of costimulatory molecules CD80, CD86, and CD83, inhibi-ted production of IL-12, and inability to activate naive Tcellsin vitro. Thus, blockade of selectins using PsL-EGFmAb will prove to bea valuable tool for the study of the molecuar mechanisms of DC maturation, as well as for the prevention and treatment of DC-mediated au-toimmunity.展开更多
Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cel...Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cells (DCs), has potent adjuvant effects that polarize responses toward Th1. With a calculated molecular weight of 54.8 kDa, Hsp70L1 is smaller in size than Hsp70 but resembles it both structurally and functionally. Hsp70L1 shares common receptors on DCs with Hsp70 and can interact with DCs, promoting DC maturation and stimulating secretion of the proinflammatory cytokines interleukin 12p70 (IL-12p70), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and the chemokines IP-10, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and normal T cell expressed and secreted (RANTES). The induction of interferon-gamma-inducible protein 10 (IP-10) secretion by Hsp70L1 is not shared by Hsp70, and other functional differences include more potent stimulation of DC IL-12p70, CC-chemokine, and CCR7 and CXCR4 expression by Hsp70L1. Immunization of mice with the hybrid peptide Hsp70L1-ovalbumin(OVA)(257-264) induces an OVA(257-264)-specific Th1 response and cytotoxic T lymphocyte (CTL) that results in significant inhibition of E.G7-OVA tumor growth. The ability of Hsp70L1 to activate DCs indicates its potential as a novel adjuvant for use with peptide immunizations; the Hsp70L1 antigen peptide hybrid may serve as a more effective vaccine for the control of cancer and infectious diseases.展开更多
Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbr...Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.展开更多
Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles o...Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.展开更多
Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.In view of their exceptional ability to present antigen and to interact...Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.In view of their exceptional ability to present antigen and to interact with T cells,DC play distinct roles in linking innate and adaptive immune responses and thus become logical targets for cancer immunotherapy.Evidences show that tumor-derived cytokines could impair DCs’biomechanics properties,which lead to inefficacy of DCs-based immune therapies.Our previous studies found that IL-10,as one of the widespread suppressive cytokines from tumor microenvironment(TME),could deteriorate DCs’motility and biomechanics properties while the underlying mechanism is unknown.In this study,CD14+monocytes were induced to differentiate into mature dendritic cells after isolation in vitro using recombinant cytokines IL-4,GM-CSF,LPS,and IFN-γ.And we also have compared the proteomic changes of mDCs treated by IL-10 and control group via two-dimensional electrophoresis combine with MALDI-TOF/TOF MS.Then we analyzed the function of differentially expressed proteins through bioinformatics methods include GO analysis that clarified the biological functions of differential proteins and KEGG analysis which enriched signal pathways of differential proteins to explore the molecular mechanism of IL-10 which has inhibitory effect on mDCs.The results showed that IL-10 significantly affected the morphology of mDCs,especially reducing the number and length of filopodia.Different expressed proteins were analyzed by two-dimensional electrophoresis combined with bioinformatics analysis to enrich for glycolytic signaling pathway,HIF-1 signaling pathway and cytoskeletal binding protein expression changes.The results of two-dimensional electrophoresis were verified by Western blot,and the results showed that the data were reliable.In addition,the intracellular ROS levels were significantly higher in mDCs treated with IL-10,validating the previously enriched HIF-1 signaling pathway.In summary,it indicated that IL-10 may interfered with the oxidative metabolic process,glycolytic metabolism,and expression of cytoskeleton-related proteins in mDCs,and disturbances in these physiological processes resulted in reduced biomechanics properties and motility of mDCs and subsequently impaired their immune functions,making DC-based tumor vaccines less effective than which we desired.Our study reveals alterations in the physiological metabolism of mDCs under IL-10 treatment from the proteome,which lays the foundation for further exploration of the altered state of mDCs in the tumor microenvironment.展开更多
Implant materials,as foreign objects to host,can cause various degrees of inflammation in most cases.The inflammation is triggered by a series of immune responses and directly impacts the tissue regeneration process,w...Implant materials,as foreign objects to host,can cause various degrees of inflammation in most cases.The inflammation is triggered by a series of immune responses and directly impacts the tissue regeneration process,which determines the outcome of tissue repair.The immune responses are complex process involving numerous immune cells and can be divide into innate immune and adaptive immune responses.Once materials are implanted,innate immune responses are activated under the mediation of several immune cells(e.g.neutrophils and macrophages),meanwhile immature dendritic cells(imDCs)are recruited to the implant sites to recognize,internalize and process antigens.Upon antigen uptake,imDCs gradually differentiate into mature dendritic cells(mDCs)and migrate to secondary lymph nodes.In the lymph nodes,mDCs present processed antigen peptides to naive T lymphocytes and activate their antigen specific proliferation,resulting in initiation of adaptive immune responses.Due to their key position in the immune system,serving to bridge innate and adaptive immunity,DCs are crucial to guiding and modulating the immune responses caused by implanted materials.Therefore,figuring out the response of DCs to implanted materials and the exact role of DCs in tissue healing processes will provide deeper insight for the rational design of biomaterials.Previous studies on the effects of implants on immune functions of DCs are mainly focused on physical and chemical properties of the materials(e.g.released chemical composition,surface chemistry,substrate stiffness and surface topography).All these factors will change the microenvironment of the tissue around implant materials,which affect the immune functions of DCs.However,the change of microenvironment not only directly derives from the physical and chemical properties of the material(intrinsic),but also indirectly results from the remodeled extracellular matrix(ECM)caused by implanted materials.When blood or tissue fluid contact with materials after implantation,proteins(e.g.fibrin and collagen)will absorb and deposit on the surface of implants,leading to a provisionally stable matrix with microporous fibrous-liked network structure.It means that the remodeled ECM can provide adhesion sites for recruited DCs and form spatial confinement.DCs,as a kind of cells that are extremely sensitive to mechanical stimuli,theoretically,can response to the mechanical stimuli coming from spatial confinement of remodeled ECM,which may lead to a series of modulations in their cell morphologies and immune functions.Then,the remodeled ECM is a non-negligible mechanical cue.However,to the best of our knowledge,there is a lack of a simple and effective model to establish the relationship between the immune functions of DCs and remodeled ECM.Most studies on the responses of DCs to implanted materials are still based on suspension culture model,which is the normal status of DCs in vitro culture systems.In addition,the processes by which DC exerts immune functions(both endocytosis and antigen presentation)are dynamically physical interaction.It means that the changes of DCs’immune functions are highly correlated with the changes of their biomechanical characteristics caused by remodeled ECM.In this work,we have found that the ECM was remodeled by a large amount of fibrin matrix deposited on the surface of implants in the early stage of the inflammations following implantation.Thus,we used non-toxic salmon fibrin hydrogels with microporous fibrous-liked network structure to mimic the deposited fibrin matrix.Then,human monocyte-derived DCs were cultured on the surface and inside of the fibrin hydrogels to mimic the different spatial confinement states of fibrin matrix.Our results indicated that cell morphologies and cytoskeleton structures of DCs were regulated by the spatial confinement of fibrin hydrogels,resulting in generating mechanical stimuli for DCs.Furthermore,we have found that the biomechanical characteristics and the immune functions of both imDCs and mDC were also modulated.Considering the changes in surface markers,secreted cytokines and biomechanical characteristics of DCs,it indicates that the tendency and magnitude of modulations were highly associated with the spatial confinement of fibrin hydrogels.This model demonstrated that mechanical stimuli deriving from spatial confinement of deposited fibrin matrix is an important factor for regulating the biomechanical characteristics and immune functions of DCs.展开更多
OBJECTIVE Ginsenoside metabolite compound K(CK)is a degradation product of ginsenoside in the intestine by bacteria.The anti-inflammatory and immunomodulatory activities of CK have been reported.This study investigate...OBJECTIVE Ginsenoside metabolite compound K(CK)is a degradation product of ginsenoside in the intestine by bacteria.The anti-inflammatory and immunomodulatory activities of CK have been reported.This study investigated whether CK exerted its immunoregulatory effect through modulation of dendritic cells(DCs)function.METHODS In vivo,severity of collegen-induced arthritis(CIA),T cells and DCs subsets,phenotype of DC were assayed by flow cytometry,CCL19 and CCL21 level in lymph nodes assayed by ELISA.In vitro,bone marrow-derived DCs from normal mice were matured with lipopolysaccharide and treated with CK for 48 h.In vivo,bone marrow-derived DCs were generated from CIA mice before and 2 weeks into CK treatment.DCs were analyzed for migration,phenotype and T-cell stimulatory capacity.RESULTS CK alleviated the severity of CIA,decreased pD Cs and mo-DCs,increased na?ve T cells in CIA mice lymph nodes,and suppressed CCL21 expression in lymph nodes.CK suppressed DCs migration induced by CCL21 and T cells-stimulatory capability of DC,down-regulated LPS-induced expression of CD80,CD86,MHCII and CCR7 on DCs.CONCLUSION This study elucidated the novel immunomodulatory property of CK via impairing function of DCs in priming T cells activation.These results provide an interesting novel insight into the potential mechanism by which CK contribute to the restoration of immunoregulation in autoimmune conditions.展开更多
Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migra...Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.展开更多
文摘Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated by PGE2 and TNF-alpha respectively. The markers of maturation and activation expressed on DCs, such as CD40, CD80, CD83, CD86, MHC-II, and the ability of antigen uptake of DCs were analyzed by flow cytometry. The proliferation of T cells co-cultured with DCs, the signaling pathways of PGE2-EP4-cAMP and TNF-alpha- TRADD-TRAF2-NF-KB in DCs were analyzed. Result Both PGE2 and TNF-alpha up-regulated the expressions of CD40, CD80, CD83, CD86, and MHC-II, decreased the antigen uptake of DCs, and DCs stimulated by PGE2 or TNF-alpha could increase T cell proliferation. CP-25 （ 10 -5, 10 -6 , 10 -7 mol/L ） decreased significantly the ex- pressions of CD40, CD80, CD83, CD86 and MHC- ]I , increased the antigen uptake of DCs, and suppressed T cell proliferation induced by DCs. PGE2 increased the expressions of EP4, NF-KB and down-regulated cAMP level of DCs. TNF-alpha could also up-regulate TNFR1, TRADD, TRAF2, and NF-KB expression of DCs. CP-25 （10^-5, 10^-6, 10^-7 mol/L） decreased the expressions of EP4 and NF-KB, increased cAMP level in DCs stimulated by PGE2. CP-25 （10^-5 10^-6 10^-7 mol/L） also could down-regulate significantly TNFR1 TRADD TRAF2 and NF-KB expression in DCs stimulated by TNF-alpha. Conclusion PGE2 and TNF-alpha could enhance DCs func- tions by mediating PGE2-EP4-cAMP pathway, TNF-alpha-TNFR1-TRADD-TRAF2-NF-KB pathway respectively. CP-25 might inhibit the function of DCs through regulating PGE2-EP4-cAMP and TNF-alpha-TNFR1-TRADD- TRAF2-NF-KB pathways.
文摘AIM: To study the effect of ginsenoside Rgl and Rhl on the anti - tumor activity of dendritic cells (DC). METHODS: Effect of Rgl and Rhl on the production of IL- 12 p40 protein was detected by ELISA, and the IL- 12 p40 mRNA level of DC was monitored by RT- PCR. Anti - tumor activity of DC- LPAK was detemnined by neutral red staining assay. RESULTS: The results of ELLSA showed that Rgl and Rhl significantly enhanced the production of IL- 12 p40 of DC. Rgl at 1 mg/L and Rhl at 100 mg/L upregulated the IL- 12 p40 mRNA level. Rgl and Rhl enhanced the anti - tumor ability of DC, induced lyrnphokine and PHA activated killer (DC-LPAK) on human papillate tumor cell line. Each dose of Rgl can obviously accelerate the eytotoxity to L929 at the E: T ratio of 5 : 1 (P<0.05,0.01 ), while only Rhl 10 mg/L enhanced the eytotoxity ability of DC- LPAK (P < 0.05). CONCLUSION: Rgl and Rhl enhanced the production of IL-12 p40. This effect may be mediated by the increase in the mRNA level. As a result, Rg1 and Rhl oromote the ability of DC to stimulate the cytotoxitie aetieity of DC - LPAK.
基金Supported by the National Technology and Research Project(2015BAD12B01-4)
文摘Dendritic cells （DCs） are bone marrow-derived professional antigen presenting cells （APCs）, they are crucial for initiation of both innate and adaptive immune responses. In this study, chicken bone marrow （chBM） cells were cultured in medium with recombinant chicken granulocyte-macrophage colony stimulating factor （rGM-CSF） and recombinant chicken interleukin-4 （rIL-4） for 7 days, displayed the typical morphology of DCs. These immature chicken bone marrow-derived DCs （chBM-DCs） showed signifcant up-regulation of the putative CD11c and of major histocompatibility complex class II （MHC II）, but CD40 and CD86 co-stimulatory molecules were almost no up-regulated. However, maturation with lipopolysaccharide （LPS）, surface expression of CD40, CD86 was greatly increased. The phagocytosis of chBM-DCs was assessed by neutral red, and the phagocytosis decreased after stimulation. In mixed lymphocyte responses （MLR）, stimulated chBM-DCs were more effective to T-cell stimulators than non-stimulated chBM-DCs. In addition, mRNA expression levels of IL-1β, IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α, CXCLi1 and CXCLi2 were assessed by real-time qPCR （qRT-PCR）, and the results showed cultured chBM-DCs could be matured to a T helper cell type 1 （Th1）-promoting phenotype by LPS stimulation.
基金Projects(2017YFE0131900,2017YFB0404500)supported by National Key Research and Development Program of ChinaProjects(91833306,91733302,62075094)supported by the National Natural Science Foundation of China+1 种基金Project(202003N4004)supported by the Ningbo Natural Science Foundation,ChinaProject(2020GXLH-Z-014)supported by the Joint Research Funds of Department of Science&Technology of Shaanxi Province and Northwestern Polytechnical University,China。
文摘A series of shape-persistent polyphenylene dendritic C_(60)derivatives as the electron transport materials were designed and synthesized via a catalyst-free Diels-Alder[4+2]cycloaddition reaction.These increasing hyperbranched scaffolds could effectively enhance the solubility;notably,both first and second generation dendrimers,C_(60)-G1 and C_(60)-G2,demonstrated more than 5 times higher solubilities than pristine C_(60).Furthermore,both simulated and experimental data proved their promising solution-processabilities as electron-transporting layers(ETLs)for perovskite solar cells.As a result,the planar p-i-n structural perovskite solar cell could achieve a maximum power conversion efficiency of 14.7%with C_(60)-G2.
文摘Dendritic cells(DCs) are professinal antigen-presenting cells with the ability to initiate primary Tcell responses. While it iswell known that inflammatory stimuli induce the functional maturation of immature DCs, whether adhesion molecule selectins regulate DCmaturation is poorly understood. Using anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb) that blocks the adhesionof P-, E-, and L-selectin, we demonstrate herein that selectins play important role in stimulating functional maturation of immature DCs.Immature DCs are generated from human cord blood CD34+hematopoietic stem/progenitor cells that were cultured in the presence of stemcell factor, Fms-like tyrosine-kinase-3 ligand, granulocyte-macrophage colony stimulating factor, and transform growth factor-β1. Whenstimulated with tumor necrosis factor-alpha(TNF-α), immature DCs differentiated into mature DCs, producing increased levels of costim-ulatory molecules and interleukin (IL)-12 and obtaining the ability to potently activate naive Tcells. Interestingly, in contrast to matureDCs derived from TNF-α-induced immature DC cultures without PsL-EGFmAb, immature DCs treated with PsL-EGFmAb for7 days werecompletely blocked their maturation, as evidenced by decreased expression of costimulatory molecules CD80, CD86, and CD83, inhibi-ted production of IL-12, and inability to activate naive Tcellsin vitro. Thus, blockade of selectins using PsL-EGFmAb will prove to bea valuable tool for the study of the molecuar mechanisms of DC maturation, as well as for the prevention and treatment of DC-mediated au-toimmunity.
文摘Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cells (DCs), has potent adjuvant effects that polarize responses toward Th1. With a calculated molecular weight of 54.8 kDa, Hsp70L1 is smaller in size than Hsp70 but resembles it both structurally and functionally. Hsp70L1 shares common receptors on DCs with Hsp70 and can interact with DCs, promoting DC maturation and stimulating secretion of the proinflammatory cytokines interleukin 12p70 (IL-12p70), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and the chemokines IP-10, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and normal T cell expressed and secreted (RANTES). The induction of interferon-gamma-inducible protein 10 (IP-10) secretion by Hsp70L1 is not shared by Hsp70, and other functional differences include more potent stimulation of DC IL-12p70, CC-chemokine, and CCR7 and CXCR4 expression by Hsp70L1. Immunization of mice with the hybrid peptide Hsp70L1-ovalbumin(OVA)(257-264) induces an OVA(257-264)-specific Th1 response and cytotoxic T lymphocyte (CTL) that results in significant inhibition of E.G7-OVA tumor growth. The ability of Hsp70L1 to activate DCs indicates its potential as a novel adjuvant for use with peptide immunizations; the Hsp70L1 antigen peptide hybrid may serve as a more effective vaccine for the control of cancer and infectious diseases.
文摘Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.
基金funded by the National Natural Science Foundation of China ( 31660258,31771014, 31860262,31570938,31260227)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412, 2016-5676,2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04 )the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.
基金funded by the National Natural Science Foundation of China ( 31660258,31771014, 31860262,11762006)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412,2016-5676, 2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04)the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.In view of their exceptional ability to present antigen and to interact with T cells,DC play distinct roles in linking innate and adaptive immune responses and thus become logical targets for cancer immunotherapy.Evidences show that tumor-derived cytokines could impair DCs’biomechanics properties,which lead to inefficacy of DCs-based immune therapies.Our previous studies found that IL-10,as one of the widespread suppressive cytokines from tumor microenvironment(TME),could deteriorate DCs’motility and biomechanics properties while the underlying mechanism is unknown.In this study,CD14+monocytes were induced to differentiate into mature dendritic cells after isolation in vitro using recombinant cytokines IL-4,GM-CSF,LPS,and IFN-γ.And we also have compared the proteomic changes of mDCs treated by IL-10 and control group via two-dimensional electrophoresis combine with MALDI-TOF/TOF MS.Then we analyzed the function of differentially expressed proteins through bioinformatics methods include GO analysis that clarified the biological functions of differential proteins and KEGG analysis which enriched signal pathways of differential proteins to explore the molecular mechanism of IL-10 which has inhibitory effect on mDCs.The results showed that IL-10 significantly affected the morphology of mDCs,especially reducing the number and length of filopodia.Different expressed proteins were analyzed by two-dimensional electrophoresis combined with bioinformatics analysis to enrich for glycolytic signaling pathway,HIF-1 signaling pathway and cytoskeletal binding protein expression changes.The results of two-dimensional electrophoresis were verified by Western blot,and the results showed that the data were reliable.In addition,the intracellular ROS levels were significantly higher in mDCs treated with IL-10,validating the previously enriched HIF-1 signaling pathway.In summary,it indicated that IL-10 may interfered with the oxidative metabolic process,glycolytic metabolism,and expression of cytoskeleton-related proteins in mDCs,and disturbances in these physiological processes resulted in reduced biomechanics properties and motility of mDCs and subsequently impaired their immune functions,making DC-based tumor vaccines less effective than which we desired.Our study reveals alterations in the physiological metabolism of mDCs under IL-10 treatment from the proteome,which lays the foundation for further exploration of the altered state of mDCs in the tumor microenvironment.
基金funded by grants from the National Natural Science Foundation of China ( 31771014, 11762006,31660258,31860262,11762006,81460254 )the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04)+1 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the Science and Technology Foundation of Guizhou Province ( 2018-1412,2016-5676,2017-5718)
文摘Implant materials,as foreign objects to host,can cause various degrees of inflammation in most cases.The inflammation is triggered by a series of immune responses and directly impacts the tissue regeneration process,which determines the outcome of tissue repair.The immune responses are complex process involving numerous immune cells and can be divide into innate immune and adaptive immune responses.Once materials are implanted,innate immune responses are activated under the mediation of several immune cells(e.g.neutrophils and macrophages),meanwhile immature dendritic cells(imDCs)are recruited to the implant sites to recognize,internalize and process antigens.Upon antigen uptake,imDCs gradually differentiate into mature dendritic cells(mDCs)and migrate to secondary lymph nodes.In the lymph nodes,mDCs present processed antigen peptides to naive T lymphocytes and activate their antigen specific proliferation,resulting in initiation of adaptive immune responses.Due to their key position in the immune system,serving to bridge innate and adaptive immunity,DCs are crucial to guiding and modulating the immune responses caused by implanted materials.Therefore,figuring out the response of DCs to implanted materials and the exact role of DCs in tissue healing processes will provide deeper insight for the rational design of biomaterials.Previous studies on the effects of implants on immune functions of DCs are mainly focused on physical and chemical properties of the materials(e.g.released chemical composition,surface chemistry,substrate stiffness and surface topography).All these factors will change the microenvironment of the tissue around implant materials,which affect the immune functions of DCs.However,the change of microenvironment not only directly derives from the physical and chemical properties of the material(intrinsic),but also indirectly results from the remodeled extracellular matrix(ECM)caused by implanted materials.When blood or tissue fluid contact with materials after implantation,proteins(e.g.fibrin and collagen)will absorb and deposit on the surface of implants,leading to a provisionally stable matrix with microporous fibrous-liked network structure.It means that the remodeled ECM can provide adhesion sites for recruited DCs and form spatial confinement.DCs,as a kind of cells that are extremely sensitive to mechanical stimuli,theoretically,can response to the mechanical stimuli coming from spatial confinement of remodeled ECM,which may lead to a series of modulations in their cell morphologies and immune functions.Then,the remodeled ECM is a non-negligible mechanical cue.However,to the best of our knowledge,there is a lack of a simple and effective model to establish the relationship between the immune functions of DCs and remodeled ECM.Most studies on the responses of DCs to implanted materials are still based on suspension culture model,which is the normal status of DCs in vitro culture systems.In addition,the processes by which DC exerts immune functions(both endocytosis and antigen presentation)are dynamically physical interaction.It means that the changes of DCs’immune functions are highly correlated with the changes of their biomechanical characteristics caused by remodeled ECM.In this work,we have found that the ECM was remodeled by a large amount of fibrin matrix deposited on the surface of implants in the early stage of the inflammations following implantation.Thus,we used non-toxic salmon fibrin hydrogels with microporous fibrous-liked network structure to mimic the deposited fibrin matrix.Then,human monocyte-derived DCs were cultured on the surface and inside of the fibrin hydrogels to mimic the different spatial confinement states of fibrin matrix.Our results indicated that cell morphologies and cytoskeleton structures of DCs were regulated by the spatial confinement of fibrin hydrogels,resulting in generating mechanical stimuli for DCs.Furthermore,we have found that the biomechanical characteristics and the immune functions of both imDCs and mDC were also modulated.Considering the changes in surface markers,secreted cytokines and biomechanical characteristics of DCs,it indicates that the tendency and magnitude of modulations were highly associated with the spatial confinement of fibrin hydrogels.This model demonstrated that mechanical stimuli deriving from spatial confinement of deposited fibrin matrix is an important factor for regulating the biomechanical characteristics and immune functions of DCs.
基金supported by National Nature Science Foundation of China(81503084,81330081,31200675,and 81173075)
文摘OBJECTIVE Ginsenoside metabolite compound K(CK)is a degradation product of ginsenoside in the intestine by bacteria.The anti-inflammatory and immunomodulatory activities of CK have been reported.This study investigated whether CK exerted its immunoregulatory effect through modulation of dendritic cells(DCs)function.METHODS In vivo,severity of collegen-induced arthritis(CIA),T cells and DCs subsets,phenotype of DC were assayed by flow cytometry,CCL19 and CCL21 level in lymph nodes assayed by ELISA.In vitro,bone marrow-derived DCs from normal mice were matured with lipopolysaccharide and treated with CK for 48 h.In vivo,bone marrow-derived DCs were generated from CIA mice before and 2 weeks into CK treatment.DCs were analyzed for migration,phenotype and T-cell stimulatory capacity.RESULTS CK alleviated the severity of CIA,decreased pD Cs and mo-DCs,increased na?ve T cells in CIA mice lymph nodes,and suppressed CCL21 expression in lymph nodes.CK suppressed DCs migration induced by CCL21 and T cells-stimulatory capability of DC,down-regulated LPS-induced expression of CD80,CD86,MHCII and CCR7 on DCs.CONCLUSION This study elucidated the novel immunomodulatory property of CK via impairing function of DCs in priming T cells activation.These results provide an interesting novel insight into the potential mechanism by which CK contribute to the restoration of immunoregulation in autoimmune conditions.
文摘Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.
