scholarly journals The Influence of Ouabain on Human Dendritic Cells Maturation

2014 ◽  
Vol 2014 ◽  
pp. 1-15 ◽  
Author(s):  
C. R. Nascimento ◽  
R. C. Valente ◽  
J. Echevarria-Lima ◽  
C. F. L. Fontes ◽  
L. de Araujo-Martins ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Lymphocyte Activation ◽  
Human Monocytes ◽  
Atpase Inhibitor ◽  
Gm Csf ◽  
Hla Dr ◽  
Distinct Category ◽  
Thymocyte Apoptosis ◽  
Human Dendritic Cells ◽  
Dc Maturation

Although known as a Na,K-ATPase inhibitor, several other cellular and systemic actions have been ascribed to the steroid Ouabain (Oua). Particularly in the immune system, our group showed that Ouabain acts on decreasing lymphocyte proliferation, synergizing with glucocorticoids in spontaneous thymocyte apoptosis, and also lessening CD14 expression and blocking CD16 upregulation on human monocytes. However, Ouabain effects on dendritic cells (DCs) were not explored so far. Considering the peculiar plasticity and the importance of DCs in immune responses, the aim of our study was to investigate DC maturation under Ouabain influence. To generate immature DCs, human monocytes were cultured with IL-4 and GM-CSF (5 days). To investigate Ouabain role on DC activation, DCs were stimulated with TNF-αfor 48 h in the presence or absence of Ouabain. TNF-induced CD83 expression and IL-12 production were abolished in DCs incubated with 100 nM Ouabain, though DC functional capacity concerning lymphocyte activation remained unaltered. Nevertheless, TNF-α-induced antigen capture downregulation, another maturation marker, occurred even in the presence of Ouabain. Besides, Ouabain increased HLA-DR and CD86 expression, whereas CD80 expression was maintained. Collectively, our results suggest that DCs respond to Ouabain maturating into a distinct category, possibly contributing to the balance between immunity and tolerance.

Residual Lymphocytes in GM-CSF and IL-15 Differentiated Monocyte-Derived Dendritic Cells Enables Cytotoxic T Lymphocyte Responses.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4907-4907
Author(s):  
Melinda Y. Hardy ◽  
Andrew J. Kassianos ◽  
Ray Wilkinson ◽  
Annelie Vulink ◽  
Derek N.J. Hart ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Small Population ◽  
Lymphoid Cells ◽  
Gm Csf ◽  
Hla Dr ◽  
Ifn Γ ◽  
Human Dendritic Cells ◽  
Ctl Responses

Abstract We investigated the capacity of IL-15 to differentiate human dendritic cells (DC) from monocytes in the presence of GM-CSF (IL-15 MoDC) and compared them with MoDC differentiated in IL-4 and GM-CSF (IL-4 MoDC) as used in many immunotherapy protocols. IL-15 MoDC expressed higher levels of CD40 and HLA-DR and importantly, induced MART-1 specific cytotoxic T lymphocyte (CTL) responses with superior lytic capacity, when compared to IL-4 MoDC. In response to activation, IL-15 MoDC secreted high levels of IFN-γbut low or no IL-12, whereas IL-4 MoDC secreted high IL-12 but low or no IFN-γ. Using an IFN-γ blocking antibody, we demonstrated that IFN-γ production by the IL-15 MoDC did not account for the superior CTL responses induced. Despite immunoselecting monocytes to greater than 97% purity prior to DC differentiation, we noticed a small population (1–2%) of CD56+ and CD3+ lymphocytes in the IL-15 MoDC preparations that were less prominent in IL-4 MoDC differentiated from the same monocytes. Removal of the residual lymphocytes from monocytes prior to differentiation into IL-15 MoDC diminished their capacity to induce CTL but did not affect the expression of HLA-DR or CD40. These data suggest that IL-15-dependent cross-talk between the small lymphoid populations present and DC, during DC differentiation from monocytes results in superior CTL priming that is independent of IL-12 and IFN-γ. Based on these results, appropriately manufactured IL-15 MoDC preparations containing defined numbers of lymphoid cells should be considered for immunotherapy.

scholarly journals Synergy between Vitamin D3and Toll-Like Receptor Agonists Regulates Human Dendritic Cell Response during Maturation

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Anne Brosbøl-Ravnborg ◽  
Bettina Bundgaard ◽  
Per Höllsberg
Keyword(s):  
Vitamin D ◽  
Dendritic Cell ◽  
Surface Expression ◽  
Blood Monocytes ◽  
Tlr Agonists ◽  
Gm Csf ◽  
Dendritic Cell Differentiation ◽  
Hla Dr ◽  
Human Dendritic Cells ◽  
Dc Maturation

Human dendritic cells (DC) can be differentiated from blood monocytes in the presence of GM-CSF and IL-4 and matured by lipopolysaccharide (LPS). Vitamin D3inhibits the maturation of human DC measured by changes in surface expression of HLA-DR, CD14, CD40, CD80, CD83, and CD86. We here examine the function of vitamin D3during DC maturation. One of the earliest changes to LPS-induced maturation was an increase in CD83 expression. Vitamin D3inhibited the increase in expression of HLA-DR, CD40, CD80, CD83, and CD86 and the decrease in expression of CD14, which was paralleled morphologically by vitamin D3-induced inhibition of dendritic cell differentiation. Vitamin D3acted in synergy with the TLR agonists LPS and peptidoglycan (PGN) in inducing IL-6, IL-8, and IL-10, whereas vitamin D3completely inhibited LPS-induced secretion of IL-12. The synergy occurred at concentrations where neither vitamin D3nor the TLR agonists alone induced measurable cytokine secretion. Both LPS and PGN enhanced the level of the vitamin D3receptor (VDR). Taken together, these data demonstrated that vitamin D3and TLR agonists acted in synergy to alter secretion of cytokines from human DC in a direction that may provide an anti-inflammatory environment.

NF-κB Blockade Induces Apoptosis and Decreases Costimulatory Molecules and IL-12 Secretion of Dendritic Cells, Altering DC Maturation: Role in Induction of Tolerance after Transplantation.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2465-2465
Author(s):  
Luis I. Sanchez Abarca ◽  
Jose A. Perez-Simon ◽  
Belen Blanco ◽  
Norma Gutierrez ◽  
Juan Mateos ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Allogeneic Transplantation ◽  
Costimulatory Molecules ◽  
Cytokine Secretion ◽  
Gm Csf ◽  
Hla Dr ◽  
New Strategy ◽  
Th1 Cytokine ◽  
Dc Maturation

Abstract In steady state, dendritic cells (DC) are in an immature status and maintain tolerance to self antigens. By contrast, if DC maturation is stimulated, as it occurs in the proinflamatory milieu induced by allogeneic transplantation, they trigger effector and memory T cells and, for this reason, they play an essential role in the development of graft versus host disease (GVHD). DC maturation and survival is dependent on NF-kB. Thus, we explored the ability of the proteosome inhibitor bortezomib (B), which prevents Nuclear Factor kB (NF-kB) activation, to induce DC apoptosis and block maturation as a new strategy to prevent GVHD. DC were obtained from PBMN after culture with GM-CSF and IL-4 and were stimulated to maturate and express costimulatory molecules (CD80, CD83, CD86, CD40L) adding TNF and LPS. Even after this stimulation, B at doses ranging from 10 to 50 nM was able to decrease costimulatory molecules as well as HLA-DR expression, as assessed by flow cytometry. In addition, at 50 nM, B increased DC apoptosis (2650 vs 1404 anex/7AAD positive events among DC cultured with or w/o B, respectively; p=0.05). Moreover, B significantly decreased the production of IL-12, while IL10 secretion was unaffected (1,4% vs 6,2% DC cultured with or w/o B secreting IL-12, respectively; p=0.01). Cytokines and proinflamatory molecules genes expression profiling was also analysed by microarrays in DC cultured with or without B. Finally, after culture with bortezomib, DC were cocultured with T lymphocytes and, interestingly, allogeneic mixed lymphocyte reaction was markedly inhibited also inducing a decrease in Th1 cytokine secretion among T cells. In conclusion, bortezomib induces apoptosis and decreases DC maturation and cytokine secretion and favours tolerance to alloantigens. This new strategy based on DC therapy supports the use of proteosome inhibitors in the management of GVHD.

scholarly journals Phenotype and functions of human dendritic cells derived from CD14+ monocyte subsets opposed to CD16 expression

2019 ◽  
Vol 18 (1) ◽  
pp. 266-276 ◽  
Author(s):  
E. R. Chernykh ◽  
T. V. Tyrinova ◽  
O. Yu. Leplina ◽  
M. A. Tikhonova ◽  
Yu. D. Kurochkina ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Suppressive Effect ◽  
Inhibitory Effect ◽  
Gm Csf ◽  
Healthy Donors ◽  
Cd16 Expression ◽  
Monocyte Subpopulations ◽  
Macrophage Colony ◽  
Human Dendritic Cells ◽  
Dc Maturation

The aim of the study was to analyze the relationship between monocyte subpopulations and phenotype/ functions of monocyte-derived dendritic cells (DCs), as well as DC sensitivity to the tolerogenic effect of dexamethasone.Materials and methods. The study included 15 healthy donors. DCs were generated by cultivating enriched fractions of CD14+ monocytes with or without CD16+cell depletion (CD16-Mo-DCs or CD16+Mo-DCs, respectively) in the presence of interferon alpha (IFNα) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Monocyte subpopulations were obtained by immunomagnetic negative selection.Results. CD16+Mo-DCs were characterized by higher percentage of mature (CD83+CD14-) and lower number of semi-mature (CD14+CD83+) cells, but were similar to CD16-Mo-DCs by HLA-DR and CD86 expression, involved in the presentation of antigens and activation of naive T-cells. and also to co-inhibitory/ tolerogenic molecules B7-H1 and TLR-2. CD16+Mo-DCs displayed higher allostimulatory activity, which was positively correlated with CD86 expression (rS = 0.69; p = 0.027) and negatively – with TLR-2 expression (rS = -0.72; p = 0.1). Allostimulatory activity of CD16-Mo-DCs was positively correlated with the number of mature CD14-CD83+DCs and semi-mature CD14+CD83+DCs. Addition of dexamethasone (10-6 M) into CD16-Mo-DCs and CD16+Mo-DCs cultures led to the delay of DC maturation, the decrease of CD86 and the increase of TLR-2 expression, as well as the increase of cells with co-inhibitory CD86- B7-H1+ phenotype that was positively correlated with the reduction of DC allostimulatory activity. The decrease of CD86+/TLR-2+ index in CD16+Mo-DC population was due to the reduction of CD86+DCs and in CD16-Mo-DC population – to the increase of TLR-2+cells. Dexamethasone possessed higher inhibitory effect on DC maturation in the CD16+Mo-DC cultures.Conclusion. CD14+ monocytes, both contained and depleted by CD16+ cells, can differentiate into DCs when cultured with IFNα. The presence of CD16+ cells in whole blood monocyte pool is associated with generation of DCs showed a more mature phenotype and higher allostimulatory activity. Both CD16- and CD16+ monocyte-derived DCs are sensitive to suppressive effect of dexamethasone. However, dexamethasone tolerogenic effect involves different mechanisms in CD16-Mo-DCs and CD16+Mo-DCs.

scholarly journals Immunological role of primary cilia of dendritic cells in human skin disease

2020 ◽  
Author(s):  
Manami Toriyama ◽  
Defri Rizaldy ◽  
Motoki Nakamura ◽  
Fumitaka Fujita ◽  
Fumihiro Okada ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Primary Cilia ◽  
Potential Role ◽  
Skin Barrier ◽  
Dependent Manner ◽  
Gm Csf ◽  
Human Immune ◽  
Human Dendritic Cells ◽  
Dc Maturation

AbstractPrimary cilia are a unique organelle, known to provide a signaling hub for variety of cell activities. Their potential role(s) in human immune homeostasis and diseases, however, have yet to be explored. Here, we show that human dendritic cells (DCs) express primary cilia-like structure. The primary cilia growth during DC proliferation by GM-CSF was shut off by DC maturation agents, suggesting the role of primary cilia to transduce proliferation signaling. PDGFRα pathway, one of proliferation signal in primary cilia, promoted DC proliferation in a dependent manner of intra-flagellar transport system. In epidermis with atopic dermatitis patients, aberrant ciliated langerhans cells and keratinocytes with showing immature state were observed that may play a potential role in inflammation and skin barrier disorder.

scholarly journals Nitration of the Pollen Allergen Bet v 1.0101 Enhances the Presentation of Bet v 1-Derived Peptides by HLA-DR on Human Dendritic Cells

PLoS ONE ◽  
2012 ◽  
Vol 7 (2) ◽  
pp. e31483 ◽  
Author(s):  
Anette C. Karle ◽  
Gertie J. Oostingh ◽  
Sonja Mutschlechner ◽  
Fatima Ferreira ◽  
Peter Lackner ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Pollen Allergen ◽  
Bet V 1 ◽  
Hla Dr ◽  
Human Dendritic Cells

scholarly journals Dendritic cells mediate the induction of polyfunctional human IL17-producing cells (Th17-1 cells) enriched in the bone marrow of patients with myeloma

Blood ◽  
2008 ◽  
Vol 112 (7) ◽  
pp. 2878-2885 ◽  
Author(s):  
Kavita M. Dhodapkar ◽  
Scott Barbuto ◽  
Phillip Matthews ◽  
Anjli Kukreja ◽  
Amitabha Mazumder ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Th17 Cells ◽  
T Helper ◽  
Tumor Bed ◽  
Distinct Lineage ◽  
Human Dendritic Cells ◽  
Antigen Presenting ◽  
Dc Maturation

Abstract IL17-producing (Th17) cells are a distinct lineage of T helper cells that regulate immunity and inflammation. The role of antigen-presenting cells in the induction of Th17 cells in humans remains to be fully defined. Here, we show that human dendritic cells (DCs) are efficient inducers of Th17 cells in culture, including antigen-specific Th17 cells. Although most freshly isolated circulating human Th17 cells secrete IL17 alone or with IL2, those induced by DCs are polyfunctional and coexpress IL17 and IFNγ (Th17-1 cells). The capacity of DCs to expand Th17-1 cells is enhanced upon DC maturation, and mature DCs are superior to monocytes for the expansion of autologous Th17 cells. In myeloma, where tumors are infiltrated by DCs, Th17 cells are enriched in the bone marrow relative to circulation. Bone marrow from patients with myeloma contains a higher proportion of Th17-1 cells compared with the marrow in preneoplastic gammopathy (monoclonal gammopathy of undetermined significance [MGUS]). Uptake of apoptotic but not necrotic myeloma tumor cells by DCs leads to enhanced induction of Th17-1 cells. These data demonstrate the capacity of DCs to induce expansion of polyfunctional IL17-producing T cells in humans, and suggest a role for DCs in the enrichment of Th17-1 cells in the tumor bed.

scholarly journals In-VitroDifferentiation of Mature Dendritic Cells From Human Blood Monocytes

1998 ◽  
Vol 6 (1-2) ◽  
pp. 25-39 ◽  
Author(s):  
Robert Gieseler ◽  
Dirk Heise ◽  
Afsaneh Soruri ◽  
Peter Schwartz ◽  
J. Hinrich Peters
Keyword(s):  
Dendritic Cells ◽  
Human Blood ◽  
T Cell Response ◽  
Blood Monocytes ◽  
Gm Csf ◽  
Hla Dr ◽  
Hla Dq ◽  
Specific Stimulation ◽  
Antigen Presenting

Representing the most potent antigen-presenting cells, dendritic cells (DC) can now be generated from human blood monocytes. We recently presented a novel protocol employing GM-CSF, IL-4, and IFN-γto differentiate monocyte-derived DCin vitro. Here, such cells are characterized in detail. Cells in culture exhibited both dendritic and veiled morphologies, the former being adherent and the latter suspended. Phenotypically, they were CD1a-/dim, CD11a+, CD11b++, CD11c+, CD14dim/-, CD16a-/dim, CD18+, CD32dim/-, CD33+, CD40+, CD45R0+, CD50+, CD54+, CD64-/dim, CD68+, CD71+, CD80dim, CD86+/++, MHC class I++/+++HLA-DR++/+++HLA-DP+, and HLA-DQ+. The DC stimulated a strong allogeneic T-cell response, and further evidence for their autologous antigen-specific stimulation is discussed. Although resembling a mature CD 11c+CD45R0+blood DC subset identified earlier, their differentiation in the presence of the Thl and Th2 cytokines IFN-γand IL-4 indicates that these DC may conform to mature mucosal DC.

Leukemic blasts fail to differentiate into mature antigen presenting cells, and may hinder dendritic cell maturation in vitro and in vivo

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 10556-10556
Author(s):  
J. Rosenblatt ◽  
R. Stone ◽  
C. Lenahan ◽  
Z. Wu ◽  
B. Vasir ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Tumor Vaccine ◽  
Leukemia Cells ◽  
Phenotypic Characteristics ◽  
Gm Csf ◽  
Maturation In Vitro ◽  
Dc Maturation

10556 Background: Dendritic cells (DC) play a key role in the development of tumor specific immune responses. Dendritic cells differentiated from leukemic blasts (LDC) are being explored as a tumor vaccine in AML. We examined the phenotypic and functional characteristics of LDC, the phenotypic characteristics of native DC in AML patients, and the effect of leukemic blasts on the phenotype of DC generated from normal donors. Methods: Leukemia blasts were isolated from peripheral blood of 24 patients with AML. LDC were generated by culturing blasts in the presence of GM-CSF, IL-4 and TNFa for 7 days. The phenotype of circulating DC1 (CD11C+/lin-) and DC2 (CD123+/ lin-) in AML patients was assessed by multichannel FACS analysis. To assess the effect of blasts on DC maturation, adherent mononuclear cells were isolated from normal donors, combined with leukemia cells in a 10:1 ratio, and cultured with GM-CSF, IL-4, and TNFa. Results: LDC demonstrate only modest expression of the costimulatory molecules CD80 and CD86 (mean expression 10% and 32%) and poorly express the maturation marker CD83 (mean expression 4%). Interferon gamma production by autologous T cells was not higher after stimulation with LDC than with blasts. LDC stimlation resulted in a 2 fold increase in both CD4+/CD25+/CD69+ (activated) and CD4+/CD25+/FOXP3+ (regulatory) T cells. Given the inability of leukemia progenitors to differentiate into phenotypically mature DC, we assessed whether leukemia cells directly inhibit differentiation of DC from normal progenitors. Expression of costimulatory molecules was decreased in DC differentiated in the presence of blasts. Mean expression of CD80, CD83, and CD86 was 16%, 2%, 83% and 49%, 10%, 99% for DCs generated in the presence or absence blasts respectively. Phenotypic characteristics of native DC in patients with AML were examined. In 3 experiments, a predominance of DC2 was seen (ratio DC2/DC1 5), and both DC1 and DC2 poorly expressed CD83 (mean expression 9% DC1, 0.9% DC2). Conclusions: LDC have phenotypic and functional deficiencies, limiting their efficacy as a tumor vaccine. Contact with leukemic blasts may inhibit DC maturation in vitro and in vivo, which may contribute to the lack of effective antitumor immunity in AML patients. No significant financial relationships to disclose.

Adenosine affects expression of membrane molecules, cytokine and chemokine release, and the T-cell stimulatory capacity of human dendritic cells

Blood ◽  
2003 ◽  
Vol 101 (10) ◽  
pp. 3985-3990 ◽  
Author(s):  
Elisabeth Panther ◽  
Silvia Corinti ◽  
Marco Idzko ◽  
Yared Herouy ◽  
Matthias Napp ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Mhc Class I ◽  
Class I ◽  
Interleukin 12 ◽  
T Helper 1 ◽  
Membrane Expression ◽  
Hla Dr ◽  
Factor Α ◽  
Human Dendritic Cells ◽  
Th1 Immune Responses

Abstract Dendritic cells (DCs) express functional purinergic type 1 receptors, but the effects of adenosine in these antigen-presenting cells have been only marginally investigated. Here, we further characterized the biologic activity of adenosine in immature DCs (iDCs) and lipopolysaccharide (LPS)–matured DCs (mDCs). Chronic stimulation with adenosine enhanced the macropinocytotic activity and the membrane expression of CD80, CD86, major histocompatibility complex (MHC) class I, and HLA-DR molecules on iDCs. Adenosine also increased LPS-induced CD54, CD80, MHC class I, and HLA-DR molecule expression in mDCs. In addition, adenosine dose-dependently inhibited tumor necrosis factor α and interleukin-12 (IL-12) release, whereas it enhanced the secretion of IL-10 from mDCs. The use of selective receptor agonists revealed that the modulation of the cytokine and cell-surface marker profile was due to activation of A2 adenosine receptor. Functionally, adenosine reduced the allostimulatory capacity of iDCs, but not of mDCs. More important, DCs matured in the presence of adenosine had a reduced capacity to induce T helper 1 (Th1) polarization of naive CD4+ T lymphocytes. Finally, adenosine augmented the release of the chemokine CCL17 and inhibited CXCL10 production by mDCs. In aggregate, the results provide initial evidence that adenosine diminishes the capacity of DCs to initiate and amplify Th1 immune responses.

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