Surface LSP1 is a phenotypic marker distinguishing human classical versus monocyte-derived dendritic cells (original) (raw)

Extensive Phenotype of Human Inflammatory Monocyte-Derived Dendritic Cells

Cells

Inflammatory monocyte-derived dendritic cells (Mo-DCs) have been described in several chronic inflammatory disorders, such as rheumatoid arthritis (RA), and are suspected to play a detrimental role by fueling inflammation and skewing adaptive immune responses. However, the characterization of their phenotype is still limited, as well as the comprehension of the factors that govern their differentiation. Here, we show that inflammatory Mo-DCs generated in vitro expressed a large and atypical panel of C-type lectin receptors, including isoforms of CD209 and CD206, CD303 and CD207, as well as intracellular proteins at their surfaces such as the lysosomal protein CD208. Combination of these markers allowed us to identify cells in the synovial fluid of RA patients with a close phenotype of inflammatory Mo-DCs generated in vitro. Finally, we found in coculture experiments that RA synoviocytes critically affected the phenotypic differentiation of monocytes into Mo-DCs, suggesting that the ...

Identification of human tissue cross-presenting dendritic cells

OncoImmunology, 2013

Dendritic cells (DCs) are responsible for the induction of immune responses to pathogens, tumors and vaccines as well as for the maintenance of self-tolerance. 1 DCs were first described in the mouse spleen and elegant studies in murine models have demonstrated that these cells are heterogeneous and functionally specialized. 1 In mice, splenic and lymphoid tissue (LT) DCs were the historical center of research interest but recent work has focused on their migratory counterparts in non-lymphoid tissues (NLTs) such as the skin, gut and lung. the in vivo counterpart of human monocyte-derived DCs are still unclear. The accessibility of the human skin enabled investigations on cutaneous DCs, containing two subsets, which are defined by the expression of CD1c/CD1a and CD14 in the dermis, in addition to epidermal Langerhans cells (LCs). 6 However, the relationships between human blood CD1c + DCs and CD141 + DCs with skin CD1c/CD1a + DCs and CD14 + DCs are uncertain. CD1c+ and CLEC9A+ DCs have been demonstrated in human lymph nodes (LNs). The phenotypic differences, nomenclature issues and 80 million years of independent evolution between mouse and human have prevented an easy translation of results from mouse DC experimentation into knowledge applicable to humans. Dalod and colleagues were the first to attempt steady-state inter-species correlation of DC subsets and suggested by transcriptomic analysis a homologous relationship between human blood CD141 + DCs and CD1c + DCs with mouse splenic CD8 + DCs and CD11b + DCs, respectively. 8 Recently, functional equivalence between human blood and splenic CD141 + DCs with mouse splenic CD8 + DCs was demonstrated. 5 However, the identity of the human tissue crosspresenting DCs, which by virtue of its location constitutes an ideal target for DC-based anticancer therapy, remained elusive. We have recently identified human tissue cross-presenting DCs and aligned the human and mouse NLT-DC networks . We developed a 12-parameter flow cytometry strategy to visualize known DC and monocyte subsets in the human blood. By applying the same analysis to cell suspension derived from the human skin, lung and liver we identified a new DC subset, CD141 + DCs, in addition to the known CD1c + DCs and CD14 + DCs, which were distinct from resident macrophages. In the skin, CD141 expression was promiscuous and was detectable on all DC subsets including CD14 + DCs, as recently described. 10 Tissue CD141 + DCs also expressed CD1c and CD1a and hence will be referred to hereafter as CD141 hi DCs, to distinguish them from blood CD141 + DCs and other DC subsets coexpressing CD141. 10 Blood CD141 + DCs are rare (< 0.1% of CD45 + cells) but tissue CD141 hi DCs seem at least 10-fold more abundant. Similar to their circulating counterparts, skin CD141 hi DCs express XCR1, TLR3, CLEC9A and CADM1 Dendritic cells (DCs) are a heterogeneous group of functionally specialized antigen-presenting cells. we recently characterized the human tissue cross-presenting DCs and aligned the human and mouse DC subsets. Our findings will facilitate the translation of murine DC studies to the human setting and aid the design of DC-based vaccine strategies for infection and cancer immunotherapy.

The mononuclear phagocyte–dendritic cell dichotomy: myths, facts, and a revised concept

Clinical and Experimental Immunology, 1996

Since Aschoff's reticuloendothelial system was abandoned a few decades ago, classification and characterization of the mononuclear phagocyte and dendritic cell systems have evolved separately or even in competition with one another. New information has now become available indicating that monocytes/macrophages and dendritic cells have a common origin in the bone marrow, and may even transdifferentiate. Morphological and functional distinctions—although valid under certain conditions—have been blurred by revelation of the versatility of monocytes/macrophages and dendritic cells in response to different contextual needs in inflammation and immunity. Monocytes/macrophages and dendritic cells share a sentinel, receptor/effector, and presentation mode, and may either activate or silence specific immune reactions. In keeping with the view of monocytes/macrophages and dendritic cells as interactive sentinels, we suggest that the mononuclear phagocyte and dendritic cell systems be replaced by the custocyte system (custos, Lat = sentinel, guard) as a unifying concept. Within the custocyte system, we recognize type I, type II, and type III custocytes. Type I and II custocytes exhibit predominance of presentation or effector/presenter interdependency, respectively, while type III custocytes are bipolar, passing through type I- and type II-like phases during their development and in inflammatory responses. The custocyte system brings into view monocytes/macrophages and dendritic cells as dynamic players in immunity and inflammation with a high degree of derivational, phenotypic, functional, and molecular plasticity.

Circulating CD2+ Monocytes Are Dendritic Cells

The Journal of Immunology

Low levels of CD2 have been described on subsets of monocytes, macrophages, and dendritic cells. CD2 is expressed on about one-third of circulating monocytes, at levels one-half log lower than on T or NK cells, representing 2–4% of PBMC. FACS analysis of CD2+ and CD2− monocytes revealed no significant difference in the expression of adhesion molecules (CD11a/b/c), class II Ags (HLA-DR, -DQ, -DP), myeloid Ags (CD13, CD14, CD33), or costimulatory molecules (CD80, CD86). Freshly isolated CD2+ and CD2− monocytes were morphologically indistinguishable by phase contrast microscopy. However, scanning electron microscopy revealed large prominent ruffles on CD2+ monocytes in contrast to small knob-like projections on CD2− monocytes. After 2 days of culture, the CD2+ monocytes largely lost CD14 expression and developed distinct dendrites, whereas the CD2− monocytes retained surface CD14 and remained round or oval. Freshly isolated CD2+ monocytes were more potent inducers of the allogeneic MLR...

New insights into the phenotype of human dendritic cell populations

Clinical & Translational Immunology, 2016

HLDA10 is the Tenth Human Leukocyte Differentiation Antigen (HLDA) Workshop. The HLDA Workshops provide a mechanism to allocate cluster of differentiation (CD) nomenclature by engaging in interlaboratory studies. As the host laboratory, we invited researchers from national and international academic and commercial institutions to submit monoclonal antibodies (mAbs) to human leukocyte surface membrane molecules, particularly those that recognised molecules on human myeloid cell populations and dendritic cells (DCs). These mAbs were tested for activity and then distributed as a blinded panel to 15 international laboratories to test on different leukocyte populations. These populations included blood DCs, skin-derived DCs, tonsil leukocytes, monocyte-derived DCs, CD34-derived DCs, macrophage populations and diagnostic acute myeloid leukaemia and lymphoma samples. Each laboratory was provided with enough mAb to perform five repeat experiments. Here, we summarise the reactivity of different mAb to 68 different cell-surface molecules expressed by human myeloid and DC populations. Submitted mAbs to some of the molecules were further validated to collate data required to designate a formal CD number. This collaborative process provides the broader scientific community with an invaluable data set validating mAbs to leukocyte-surface molecules.

Monocyte-Derived Dendritic Cells Isolation and Phenotypic Characterization

2020

Dendritic cell (DC) is the most powerful inducers and regulators of immune responses, responsible for interaction within the immune system. The ability of DC to induce or regulate/suppress immune responses led to attention in their immunotherapeutic use in various disease types. The aim of the presented study was to generate in vitro CD14+ enriched peripheral blood monocytes (PBMCs) following the culture with human monocyte-derived dendritic cell serum-free differentiation media over ten days with phenotypical analysis and morphological identification for functional studies. In vitro, peripheral blood of human donors samples were collected for the purification of blood CD14+ monocytes that represent the most common origin for DC precursors based on ficoll density gradient separation and human monocyte-derived dendritic cell enriched differentiation medium. After PBMCs isolation, the cell viability, cell yield were determined, the monocyte-derived dendritic cell surface marker expres...

Evaluation of Phagocytic Activity in Human Monocyte-Derived Dendritic Cells

Journal of Aerosol Medicine, 2001

antigens. Their potency in antigen presentation vis-à-vis reported low or lack of ability to phagocytize particulate matter has limited our understanding of the role that they play in inducing immunity to particulate antigens. One hypothesis is that dendritic cells may possess a high phagocytic capacity when immature and located in peripheral tissues, which they lose on maturation. Our goal was to characterize the phagocytic capacity in human immature dendritic cells. The phagocytic capacity of human monocyte-derived immature dendritic cells was studied by morphological and morphometric means, and compared to that of professional phagocytes, human alveolar macrophages, their progenitors, the peripheral blood monocytes, and mature dendritic cells. Phagocytic index (proportion of phagocytic cells) was decreased by 42.8% (immature dendritic cells) and 74.2% (mature dendritic cells) with respect to monocytes. Similarly, the phagocytic index was decreased by 46.5% (immature dendritic cells) and 75.9% (mature dendritic cells) with respect to macrophages. Volume density of phagocytized particles was decreased by 76.1% (immature dendritic cells) and 96.7% (mature dendritic cells) with respect to the monocytes. However, volume density was decreased by 34.3% (immature dendritic cells) and 91% (mature dendritic cells) with respect to alveolar macrophages. These results show that human monocyte-derived immature dendritic cells possess a phagocytic capacity that is lower than that of peripheral blood monocytes and alveolar macrophages but higher than that of mature dendritic cells.

In-Vitro Differentiation of Mature Dendritic Cells From Human Blood Monocytes

Developmental Immunology, 1998

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.

In-VitroDifferentiation of Mature Dendritic Cells From Human Blood Monocytes

Developmental Immunology, 1998

Surface LSP1 is a phenotypic marker distinguishing human classical versus monocyte-derived dendritic cells (original) (raw)

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