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.

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.

Abstract 1046: The ApoA-I Mimetic Peptide, 4F, Inhibits LPS-induced Inflammatory Reactions Through Dysregulation Of The NFkB Activation Pathway.

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Geeta Datta ◽  
C. R White ◽  
Manjula Chaddha ◽  
M. N Palgunachari ◽  
G. M Anantharamaiah ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Inflammatory Responses ◽  
Macrophage Phenotype ◽  
Blood Monocytes ◽  
Potent Inducer ◽  
Mimetic Peptide ◽  
Gm Csf ◽  
Dendritic Cell Differentiation ◽  
Anti Inflammatory ◽  
Pre Treatment

Background: The apoA-I mimetic peptide 4F possesses potent anti-inflammatory properties. We previously reported that 4F inhibits lipopolysaccharide (LPS)-induced cytokine, chemokine and adhesion molecule expression in vivo and in vitro . Other data show that 4F reduces macrophage content in atheroma in susceptible mice. Objective: In this study, we explored anti-inflammatory mechanisms of 4F action under basal conditions and in the presence of LPS using THP1 monocytes/macrophages and primary human blood monocytes. Methods and Results: Blood monocytes isolated by Ficoll and CD14+ MACS separation were analyzed by FACS for CD14, TLR-4 and p-NFκB expression. Cytokines, chemokines and adhesion molecules were measured by the Searchlight multi-analyte analysis (Pierce, MA, USA). 4F treatment (3 days) of both THP-1 and blood monocytes enhanced the secretion of GM-CSF (25-fold), a potent inducer of monocyte to dendritic cell differentiation. 4F also inhibited (by 80%) the PMA-induced phagocytosis of FITC-labeled beads by THP-1 cells. In subsequent studies, effects of 4F on LPS-induced inflammatory responses in monocytes were tested. Pre-treatment with 4F significantly inhibited LPS-induced MCP-1 expression (by 50%) in both THP-1 cells and blood monocytes. Under these conditions, the expression of the anti-inflammatory cytokine IL-10 was increased by 200%. The effects of 4F on the activation of NFκB, a downstream mediator of LPS action, were studied using flow cytometry. 4F pre-treatment significantly inhibited LPS-induced NFκB phosphorylation (NFκBp65) compared to vehicle treatment. Protective effects of 4F in LPS-treated monocytes were associated with a 50% reduction in the expression of TLR-4 and CD14, cell surface receptors for LPS. Conclusions: These data provide evidence that 4F modulates functional properties of monocytes and show for the first time that 4F inhibits LPS-induced inflammatory responses by inhibiting the expression of CD14, TLR-4 and phosphorylation of NFκB. Enhanced GM-CSF expression and inhibition of monocyte phagocytic activity also suggest that 4F may favor the differentiation of monocytes to a dendritic cell rather than a macrophage phenotype.

scholarly journals Cytokines in chronic inflammatory arthritis. IV. Granulocyte/macrophage colony-stimulating factor-mediated induction of class II MHC antigen on human monocytes: a possible role in rheumatoid arthritis.

1989 ◽  
Vol 170 (3) ◽  
pp. 865-875 ◽  
Author(s):  
J M Alvaro-Gracia ◽  
N J Zvaifler ◽  
G S Firestein
Keyword(s):  
Rheumatoid Arthritis ◽  
Class Ii ◽  
Tnf Alpha ◽  
Blood Monocytes ◽  
Ifn Gamma ◽  
Normal Peripheral Blood ◽  
Gm Csf ◽  
Hla Dr ◽  
Class Ii Mhc ◽  
Hla Dq

Granulocyte/macrophage CSF (GM-CSF) has recently been identified in rheumatoid arthritis (RA) synovial effusions. To study a potential role for GM-CSF and other cytokines on the induction of HLA-DR expression on monocytes and synovial macrophages, we analyzed the relative ability of recombinant human cytokines to induce the surface expression of class II MHC antigens on normal peripheral blood monocytes by FACS analysis. GM-CSF (800 U/ml) (mean fluorescence channel 2.54 +/- 0.33 times the control, p less than 0.001) and IFN-gamma (100 U/ml) (5.14 +/- 0.60, p less than 0.001) were the most potent inducers of HLA-DR. TNF-alpha and IL-4 also increased HLA-DR expression, although to a lesser degree [1.31 +/- 0.06 (p less than 0.02) and 1.20 +/- 0.03 (p less than 0.01), respectively]. IL-1 (40 U/ml), IL-2 (10 ng/ml), IL-3 (50 U/ml), IL-6 (100 U/ml), and CSF-1 (1,000 U/ml) did not affect surface HLA-DR density. GM-CSF also increased HLA-DR mRNA expression and surface HLA-DQ expression, but decreased CD14 (a monocyte/macrophage antigen) expression. The effect of GM-CSF on HLA-DR was not mediated by the generation of IFN-gamma in vitro because it was not blocked by anti-IFN-gamma mAb. GM-CSF was additive with IL-4 and low amounts (less than 3 U/ml) of IFN-gamma and synergistic with TNF-alpha. Because we have recently reported that supernatants of cultured RA synovial cells produce a non-IFN-gamma factor that induces HLA-DR on monocytes, we then attempted to neutralize this factor with specific anti-GM-CSF mAb. Four separate synovial tissue supernatants were studied, and the antibody neutralized the HLA-DR-inducing factor in each (p less than 0.01).

scholarly journals HLA-DR regulation and the influence of GM-CSF on transcription, surface expression and shedding

2004 ◽  
pp. 126-136 ◽  
Author(s):  
Sara E Perry ◽  
Sobhy M Mostafa ◽  
Richard Wenstone ◽  
Alan Shenkin ◽  
Paul J McLaughlin
Keyword(s):  
Surface Expression ◽  
Gm Csf ◽  
Hla Dr

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.

scholarly journals Echinococcus granulosus Antigen B Impairs Human Dendritic Cell Differentiation and Polarizes Immature Dendritic Cell Maturation towards a Th2 Cell Response

2007 ◽  
Vol 75 (4) ◽  
pp. 1667-1678 ◽  
Author(s):  
Rachele Riganò ◽  
Brigitta Buttari ◽  
Elisabetta Profumo ◽  
Elena Ortona ◽  
Federica Delunardo ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Echinococcus Granulosus ◽  
Therapeutic Option ◽  
Costimulatory Molecule ◽  
Lower Percentage ◽  
Th2 Response ◽  
Monocyte Differentiation ◽  
Dendritic Cell Differentiation ◽  
Antigen B ◽  
Dc Maturation

ABSTRACT Despite inducing a strong host cellular and humoral immune response, the helminth Echinococcus granulosus is a highly successful parasite that develops, progresses, and ultimately causes chronic disease. Although surgery remains the preferred therapeutic option, pharmacological research now envisages antihelminthic strategies. To understand the mechanisms that E. granulosus uses to escape host immunosurveillance and promote chronic infection, we investigated how two hydatid cyst components, purified antigen B (AgB) and sheep hydatid fluid (SHF), act on host dendritic cell (DC) differentiation from monocyte precursors and how they influence maturation of DC that have already differentiated. We evaluated the immunomodulatory potential of these antigens by performing immunochemical and cytofluorimetric analyses of monocyte-derived DCs from healthy human donors. During monocyte differentiation, AgB and SHF downmodulated CD1a expression and upregulated CD86 expression. Compared with immature DCs differentiated in medium alone (iDCs), AgB- and SHF-differentiated cells stimulated with lipopolysaccharide included a significantly lower percentage of CD83+ cells (P < 10−4) and had weaker costimulatory molecule expression. When stimulated with AgB and SHF, iDCs matured and primed lymphocytes towards the Th2 response typical of E. granulosus infection. SHF and particularly AgB reduced the production of interleukin-12p70 (IL-12p70) and tumor necrosis factor alpha in lipopolysaccharide-stimulated iDCs. Anti-IL-10 antibodies increased the levels of IL-12p70 secretion in AgB- and SHF-matured DCs. AgB and SHF induced interleukin-1 receptor-associated kinase phosphorylation and activated nuclear factor-κB, suggesting that Toll-like receptors could participate in E. granulosus-stimulated DC maturation. These results suggest that E. granulosus escapes host immunosurveillance in two ways: by interfering with monocyte differentiation and by modulating DC maturation.

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.

Sign in / Sign up

Export Citation Format

Share Document