Endothelial Cell-Derived Microparticules (EMPs) Can Induce In Vitro Maturation of Both Plasmocytoid and Myeloid Dendritic Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1273-1273
Author(s):  
Francine Garnache-Ottou ◽  
Fanny Angelot ◽  
Sabeha Biichle ◽  
Joel PLumas ◽  
Francoise Dignat George ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Endothelial Cell ◽  
Cell Lineage ◽  
Annexin V ◽  
Negative Control ◽  
Dose Effect ◽  
Blood Monocytes ◽  
Myeloid Dendritic Cells ◽  
Luminex Technology

Abstract The implication of plasmocytoid dendritic cells (pDCs) in transplantation is more and more reported, especially because of their tolerogenic properties supported by the expansion of regulatory T cells. Endothelial microparticles (EMPs) are plasma membrane-derived vesicules shed from endothelial stimulation involved in physiological and pathological processes, including intercellular communication, hemostasis and immunity. However, the interaction between DC and EMP are not well-known, especially in the context of alloreactivity. Therefore, the aims of the current study were to explore the DC/EMP interactions, more specifically the capacity of EMPs to modulate the maturation of DC. Firstly, we validated an original co-culture model involving a tumoral pDC lineage (GEN 2.2) and EMPs generated in a standard fashion from an endothelial cell lineage (HMEC U608), stimulated or not with TNF-α . EMPs was analyzed by FACS (FITC labelled annexin V) and supernatant resulting from the last wash of EMPs was used as negative control. By incubating increasing amounts of EMPs we showed that EMPs can lead to GEN pDC maturation in a dose- effect manner as evidenced by a significative increase in the HLA-DR and CD83 expression, compared to the negative controls or, of importance, compared to the maturation with CPG-A (ligand of TLR9 mimicking bacterial DNA), CD40L or R848 (ligand of TLR7 mimicking viral RNA). To confirm and expand these findings, we then cocultured with EMPs (and similar controls) circulating pDC obtained by an anti-BDCA-4 MoAb-mediated positive selection of healthy donors PBMC , and myeloid DC (mDC) obtained from peripheral blood monocytes cultured in the presence of GM-CSF and IL-4. Maturation of both normal circulating pDC and mDC occurred in the presence of EMPs and once again, in a dose-dependant fashion. The cytokine secretion pattern of the EMPs- matured mDC and pDC, determined by the Luminex technology, was as expected for such cell populations. The EMPs matured pDC secreted IL-1, IL-6, IFN-α , IL-12 and IL-10 but not TGF-β . In contrast, the EMPs matured mDC secreted mainly proinflammatory cytokines : IL-1, IL-6 and IL-8 but not IL-10. Overall, our studies demonstrate that EMPs can induce the maturation of both pDC and mDC. Such endothelial microparticules, generated by inflammatory and hemostatic disorders, could play a role in post-transplant immune responses. Determining the factors resulting in EMPs-induced pDC vs mDC maturation may open the possibility for EMPs-related immuno-intervention.

Receptor-type protein tyrosine phosphatase gamma (PTPγ), a new identifier for myeloid dendritic cells and specialized macrophages

Blood ◽  
2006 ◽  
Vol 108 (13) ◽  
pp. 4223-4231 ◽  
Author(s):  
Daniele Lissandrini ◽  
William Vermi ◽  
Marzia Vezzalini ◽  
Silvano Sozzani ◽  
Fabio Facchetti ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Protein Tyrosine Phosphatase ◽  
Human Peripheral Blood ◽  
Tyrosine Phosphatase ◽  
Advanced Pancreatic Cancer ◽  
Lymphoid Tissues ◽  
Blood Monocytes ◽  
Myeloid Dendritic Cells ◽  
Protein Tyrosine

AbstractProtein tyrosine phosphatase (PTPγ) is a receptor-like molecule with a known role in murine hematopoiesis. We analyzed the regulation of PTPγ expression in the human hematopoietic system, where it was detected in human peripheral blood monocytes and dendritic cells (DCs) of myeloid and plasmacytoid phenotypes. Its expression was maintained during in vitro monocyte differentiation to dendritic cells (moDC) and was further increased after maturation with bacterial lipopolysaccharide (LPS), CD40L, and TNFα. But PTPγ was absent when monocytes from the same donor were induced to differentiate in macrophages. B and T lymphocytes did not express PTPγ. Rather, PTPγ mRNA was expressed in primary and secondary lymphoid tissues, and the highest expression was in the spleen. PTPγ was detected by immunohistochemistry in subsets of myeloid-derived DCs and specialized macrophages (tingible bodies, sinus and alveolar macrophages). Classic macrophages in infective or reactive granulomatous reactions did not express PTPγ. Increased PTPγ expression was associated with a decreased ability to induce proliferation and interferon-γ secretion in T cells by moDCs from patients with advanced pancreatic cancer. Taken together, these results indicate that PTPγ is a finely regulated protein in DC and macrophage subsets in vitro and in vivo.

scholarly journals Phosphate-modified CpG oligonucleotides induce in vitro maturation of human myeloid dendritic cells

2020 ◽  
Vol 24 (6) ◽  
pp. 653-660
Author(s):  
A. A. Ostanin ◽  
O. Y. Leplina ◽  
E. A. Burakova ◽  
T. V. Tyrinova ◽  
A. A. Fokina ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Cpg Odn ◽  
Blood Monocytes ◽  
Myeloid Dendritic Cells ◽  
Gm Csf ◽  
Cpg Oligodeoxynucleotides ◽  
Cpg Oligonucleotides ◽  
Cpg Odns ◽  
Dc Maturation

Myeloid dendritic cells (DCs) play an important role in the immune response; therefore, the search for compounds that can effectively activate DCs is a needful goal. This study was aimed to investigate the effect of synthetic CpG oligodeoxynucleotides (CpG-ODN) on the maturation and allostimulatory activity of myeloid DCs in comparison with other PAMP and DAMP molecules. For the research, we synthesized known CpG-ODN class C (SD-101 and D-SL03) containing thiophosphate internucleotide groups, and their original phosphate-modified analogues (SD-101M and D-SL03M) with mesylphosphoramide internucleotide groups (M = μ-modification). The effects of CpG-ODN and other activators were evaluated on DCs generated from blood monocytes in the presence of GM-CSF and IFN-α (IFN-DC) or IL-4 (IL4-DC). Evaluation of the intracellular TLR-9 expression showed that both types of DCs (IFN-DC and IL4-DC) contained on average 52 and 80 % of TLR-9-positive cells, respectively. The CpG-ODNs studied enhanced the allostimulatory activity of IFN-DCs, and the effect of μ-modified CpG-ODNs was higher than that of CpG-ODNs with thiophosphate groups. The stimulating effect of CpG-ODN at a dose of 1.0 μg/ml was comparable (for D-SL03, D-SL03M, SD-101) with or exceeded (for SD-101M) the effect of LPS at a dose of 10 μg/ml. At the same time, IFN-DCs were characterized by greater sensitivity to the action of CpG-ODNs than IL4-DCs. The enhancement of DC allostimulatory activity in the presence of CpG-ODNs was associated with the induction of final DC maturation, which was confirmed by a significant decrease in the number of CD14+DC, an increase in mature CD83+DC and a trend towards an increase in CD86+DC. Interestingly, the characteristic ability of LPS to enhance the expression of the co-stimulatory molecule OX40L on DCs was revealed only for the μ-analogue SD-101M. In addition, CpG-ODNs (SD-101 and SD-101M) had a stimulatory effect on IFN-γ production comparable to the action of LPS. The data obtained indicate a stimulating effect of CpG-ODN on the maturation and allostimulatory activity of human myeloid DCs, which is more pronounced for μ-modified analogs.

scholarly journals Identification of an Arsenic-Sensitive Block to Primate Lentiviral Infection of Human Dendritic Cells

2007 ◽  
Vol 81 (21) ◽  
pp. 12086-12090 ◽  
Author(s):  
Marjorie Pion ◽  
Romaine Stalder ◽  
Rafael Correa ◽  
Bastien Mangeat ◽  
Greg J. Towers ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Cell Lineage ◽  
Myeloid Dendritic Cells ◽  
Immunodeficiency Virus ◽  
Lentiviral Infection ◽  
Antiviral Activities ◽  
Human Dendritic Cells ◽  
Hiv 1

ABSTRACT Dendritic cells are central to the early events of human immunodeficiency virus type 1 (HIV-1) transmission, but HIV-1 infects dendritic cells inefficiently in vitro compared to activated CD4+ T cells. There is a strong postentry restriction of HIV-1 infection in dendritic cells, partly mediated by the cellular restriction factor APOBEC3G. Here, we reveal that arsenic trioxide markedly increases HIV infection of immature and mature dendritic cells as well as blood-derived myeloid dendritic cells in an APOBEC3G- and TRIM5α-independent way. Our data suggest the presence of powerful, arsenic-sensitive antiviral activities in primary human immune cells of the dendritic cell lineage.

scholarly journals IN LABORATORY GENERATION AND MATURATION OF HUMAN MONOCYTE-DERIVED DENDRITIC CELLS FOR CANCER IMMUNOTHERAPY

2020 ◽  
pp. 46-52
Author(s):  
KANCHAN K. MISHRA ◽  
SUMIT BHARADVA ◽  
MEGHNAD G. JOSHI ◽  
ARVIND GULBAKE
Keyword(s):  
Dendritic Cells ◽  
Immune Responses ◽  
Gradient Centrifugation ◽  
Critical Role ◽  
Human Monocyte ◽  
Blood Monocytes ◽  
Adaptive Immune ◽  
Immunodeficiency Virus ◽  
Adaptive Immune Systems

Dendritic cells (DCs) play a critical role in the regulation of adaptive immune responses, furthermore they act as a bridge between the innate and the adaptive immune systems they have been ideal candidates for cell-based immunotherapy of cancers and infections in humans. The first reported trial using DCs in 1995, since they have been used in trials all over the world for several of indications, including cancer and human immunodeficiency virus infection. Generally, for in vitro experiments or for DCs vaccination monocyte-derived dendritic cells (moDCs) were generated from purified monocytes that isolated from peripheral blood by density gradient centrifugation. A variety of methods can be used for enrichment of monocytes for generation of clinical-grade DCs. Herein we summarized up to date understanding of systems and inputs used in procedures to differentiate DCs from blood monocytes in vitro.

scholarly journals Human macrophage maturation and heterogeneity: analysis with a newly generated set of monoclonal antibodies to differentiation antigens

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1257-1264 ◽  
Author(s):  
R Andreesen ◽  
KJ Bross ◽  
J Osterholz ◽  
F Emmrich
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Surface ◽  
Cell Lineage ◽  
Culture Conditions ◽  
Human Macrophage ◽  
Blood Monocytes ◽  
Differentiation Antigens

We have analyzed the expression of late differentiation antigens during terminal in vitro maturation of human macrophages (M phi) from blood monocytes (MO) in comparison to their distribution among mature M phi residing in various tissue sites. By immunizing mice with M phi derived from blood MO by culture on hydrophobic Teflon foils, monoclonal antibodies (mAbs) were developed (MAX.1, MAX.2, MAX.3, MAX.11) that reacted with lineage-restricted differentiation antigens. These antigens were expressed exclusively on M phi or were markedly increased after in vitro differentiation. The only overlap to another hemopoietic cell lineage was observed with MAX.3, which is shared by platelets and megakaryocytes. In the course of M phi maturation in vitro, the MAX.1 and MAX.3 antigens are detected within the cytoplasm two days before they appear on the cell surface. In contrast, the MAX.11 antigen is expressed simultaneously in the cytoplasm and at the cell surface, is found in varying degrees on a minor portion of blood MO and U937 cells, and is expressed rapidly at high density during early M phi differentiation in vitro. Among conventional mAbs that do not react with MO we found those against the transferrin (TF)-receptor, the BA-2, and the PCA1 antigen to label M phi. M phi matured in vivo and isolated from body fluids were positive with some but not all MAX mAbs. Distinctive patterns were observed with pulmonary M phi, exudate M phi from pleural and peritoneal effusions, synovial fluids, and early lactation milk. M phi from the alveolar space, for example, constantly expressed the MAX.2 antigen but not the MAX.3 antigen. Pleural effusion M phi, however, did not react with the MAX.1 mAb, but in most cases, it did react with the MAX.3 mAb. The detection of novel differentiation antigens, all expressed on monocyte-derived M phi but differently expressed on site-specific M phi in situ, underlines the remarkable heterogeneity among human M phi. The expression of these antigens is flexible because those MAX antigens that were not expressed in situ could be induced if cells from distinct tissue sites were cultured in vitro for several days. MAX mAbs may be of potential value to study both the sequential stages of maturation within the M phi lineage as well as differential developments induced by various culture conditions in parallel to environmental factors in vivo.

scholarly journals Whole Transcriptome Analysis of Myeloid Dendritic Cells Reveals Distinct Genetic Regulation in Patients with Allergies

2020 ◽  
Vol 21 (22) ◽  
pp. 8640
Author(s):  
Kijeong Lee ◽  
Mi-Ryung Han ◽  
Ji Woo Yeon ◽  
Byoungjae Kim ◽  
Tae Hoon Kim
Keyword(s):  
Dendritic Cells ◽  
Transcriptome Sequencing ◽  
Genetic Regulation ◽  
Genetic Alterations ◽  
Limited Information ◽  
Blood Monocytes ◽  
Sequencing Data ◽  
Myeloid Dendritic Cells ◽  
Th2 Immune Response ◽  
Whole Transcriptome

Dendritic cells (DCs) play critical roles in atopic diseases, orchestrating both innate and adaptive immune systems. Nevertheless, limited information is available regarding the mechanism through which DCs induce hyperresponsiveness in patients with allergies. This study aims to reveal novel genetic alterations and future therapeutic target molecules in the DCs from patients with allergies using whole transcriptome sequencing. Transcriptome sequencing of human BDCA-3+/CD11c+ DCs sorted from peripheral blood monocytes obtained from six patients with allergies and four healthy controls was conducted. Gene expression profile data were analyzed, and an ingenuity pathway analysis was performed. A total of 1638 differentially expressed genes were identified at p-values < 0.05, with 11 genes showing a log2-fold change ≥1.5. The top gene network was associated with cell death/survival and organismal injury/abnormality. In validation experiments, amphiregulin (AREG) showed consistent results with transcriptome sequencing data, with increased mRNA expression in THP-1-derived DCs after Der p 1 stimulation and higher protein expression in myeloid DCs obtained from patients with allergies. This study suggests an alteration in the expression of DCs in patients with allergies, proposing related altered functions and intracellular mechanisms. Notably, AREG might play a crucial role in DCs by inducing the Th2 immune response.

Artificial phosphatidylserine liposome mimics apoptotic cells in inhibiting maturation and immunostimulatory function of murine myeloid dendritic cells in response to 1-chloro-2,4-dinitrobenze in vitro

2007 ◽  
Vol 299 (7) ◽  
pp. 327-336 ◽  
Author(s):  
Dongmei Shi ◽  
Meng Fu ◽  
Pinshen Fan ◽  
Wei Li ◽  
Xinhui Chen ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Apoptotic Cells ◽  
Myeloid Dendritic Cells

Phenotype and function of myeloid dendritic cells derived from African green monkey blood monocytes

2006 ◽  
Vol 308 (1-2) ◽  
pp. 138-155 ◽  
Author(s):  
Lorenzo Mortara ◽  
Mickaël J.-Y. Ploquin ◽  
Abdourahmane Faye ◽  
Daniel Scott-Algara ◽  
Bruno Vaslin ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
African Green Monkey ◽  
Blood Monocytes ◽  
Myeloid Dendritic Cells ◽  
Green Monkey ◽  
And Function

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.

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