Veiled Cells Research Articles

Characterization of human afferent lymph dendritic cells from seroma fluids

Event Abstract Back to Event Characterization of human afferent lymph dendritic cells from seroma fluids Barbara Morandi1*, Irene Bonaccorsi2, 3, Mario Mesiti4, Romana Conte5, Gregorio Costa2, 3, Raffaella Iemmo3, Stefania Martini5, Soldano Ferrone6, Claudia Cantoni7, Maria Cristina Mingari1, 5, LORENZO MORETTA7 and Guido Ferlazzo2, 3* 1 Department of Experimental Medicine Universiy of Genoa, Italy 2 Department of Human Pathology, University of Messina, Italy, Italy 3 Cellular Therapy Program, University Hospital - A.O.U. Policlinico, Messina, Italy, Italy 4 Breast Cancer Unit, Carmona Home Care, Messina, Italy, Italy 5 IRCCS A.O.U. San Martino-IST-Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy, Italy 6 Department of Surgery, Massachusetts General Hospital, Boston, MA, United States 7 Istituto G.Gaslini, Genoa, Italy, Italy Dendritic cells (DCs) migrate from peripheral tissues to secondary lymphoid organs (SLO) through afferent lymph. Due to limitations in investigating human lymph, DCs flowing in afferent lymph have not been properly characterized in humans until now. In this study, DCs contained in seroma, an accrual of human afferent lymph occurring after lymph node surgical dissection, were isolated and analyzed in detail. Two main DC subsets were identified in seroma corresponding to the migratory DC subsets described in lymph nodes, i.e. CD14+ and CD1a+. The latter also included CD1abright Langherans cells. The two subsets appeared to share the same precursor and be developmentally related; both of them spontaneously released high levels of TGF and displayed similar T cell activating properties. On the other hand, they differed in membrane molecule expression, including TLRs, in their phagocytic activity and in the expression of proteins involved in antigen processing and presentation. It is worth noting that while both subsets were detected in seroma in the post-surgical inflammatory phase, only CD1a+ DCs migrated via afferent lymph in steady state conditions. Therefore, the large amount of DCs contained in seroma fluids allowed us a proper characterization of human DCs migrating via afferent lymph. Keywords: Dendritic Cells, veiled cells, human lymph, afferent lymph, dendritic cell subsets Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Adaptive Immunity Citation: Morandi B, Bonaccorsi I, Mesiti M, Conte R, Costa G, Iemmo R, Martini S, Ferrone S, Cantoni C, Mingari M, MORETTA L and Ferlazzo G (2013). Characterization of human afferent lymph dendritic cells from seroma fluids. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00967 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 28 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Barbara Morandi, Department of Experimental Medicine Universiy of Genoa, Genoa, Italy, morandibarb@yahoo.it Prof. Guido Ferlazzo, Department of Human Pathology, University of Messina, Italy, messina, Italy, guido.ferlazzo@unime.it Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Barbara Morandi Irene Bonaccorsi Mario Mesiti Romana Conte Gregorio Costa Raffaella Iemmo Stefania Martini Soldano Ferrone Claudia Cantoni Maria Cristina Mingari LORENZO MORETTA Guido Ferlazzo Google Barbara Morandi Irene Bonaccorsi Mario Mesiti Romana Conte Gregorio Costa Raffaella Iemmo Stefania Martini Soldano Ferrone Claudia Cantoni Maria Cristina Mingari LORENZO MORETTA Guido Ferlazzo Google Scholar Barbara Morandi Irene Bonaccorsi Mario Mesiti Romana Conte Gregorio Costa Raffaella Iemmo Stefania Martini Soldano Ferrone Claudia Cantoni Maria Cristina Mingari LORENZO MORETTA Guido Ferlazzo PubMed Barbara Morandi Irene Bonaccorsi Mario Mesiti Romana Conte Gregorio Costa Raffaella Iemmo Stefania Martini Soldano Ferrone Claudia Cantoni Maria Cristina Mingari LORENZO MORETTA Guido Ferlazzo Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. 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Bovine mammary dendritic cells: A heterogeneous population, distinct from macrophages and similar in phenotype to afferent lymph veiled cells

Dendritic cells (DC) are a heterogeneous population of professional antigen presenting cells and are potent stimulators of naïve T-cells. However, there is little previous research describing DC in bovine mammary tissue, primarily because of the difficulty distinguishing these cells from macrophages, which possess a similar phenotype. Using immunohistofluorescence and a combination of markers (MHC-II, CD205, CD11c), DC were localized in the bovine mammary gland and supramammary lymph node. In mammary tissue DC were found within the alveolar epithelium and within the intralobular connective tissue. In the lymph node DC were found on the periphery of B-cell areas, in the cortex, and among T-cells in the paracortex and medulla. DC in mammary parenchyma and supramammary lymph nodes were quantified and further characterized using flow cytometry. DC were CD11c(hi), CD14(lo) cells that expressed MHC-II and CD205. DC could be distinguished from macrophages based on their low CD14 expression. This research provides a better understanding of mammary gland immunology, while potentially aiding in the targeting of antigens to mucosal DC for vaccine development.

Tumoral aspects of plasmacytoid dendritic cells: What do we know in 2009?

Plasmacytoid dendritic cell (PDC) is a Th2-type dendritic cell precursor. It is an important professional effector cell characterized by its capacities to produce large amount of alpha interferon and to differentiate into dendritic cell. PDCs are scarce in normal condition. They circulate in the blood as veiled cells and enter the lymph node and mucosal site in response to immune stimulation. Besides the recent and wide-open field of the implication of PDCs in inflammatory diseases and in the microenvironment of solid or lymphoid tumours it has also been observed that PDCs can also be tumoral cells. In this paper, the authors present the different tumours thought to be originating from tumoral PDCs. To date, two kinds of tumoral conditions are recognized: the so-called PDCs proliferations in patients with myeloid disorders and the blastic plasmacytoid dendritic cell neoplasm. These two entities are exposed and discussed.

Human embryonic stem cell (hES) derived dendritic cells are functionally normal and are susceptible to HIV-1 infection

BackgroundHuman embryonic stem (hES) cells hold considerable promise for cell replacement and gene therapies. Their remarkable properties of pluripotency, self-renewal, and tractability for genetic modification potentially allows for the production of sizeable quantities of therapeutic cells of the hematopoietic lineage. Dendritic cells (DC) arise from CD34+ hematopoietic progenitor cells (HPCs) and are important in many innate and adaptive immune functions. With respect to HIV-1 infection, DCs play an important role in the efficient capture and transfer of the virus to susceptible cells. With an aim of generating DCs from a renewable source for HIV-1 studies, here we evaluated the capacity of hES cell derived CD34+ cells to give rise to DCs which can support HIV-1 infection.ResultsUndifferentiated hES cells were cultured on S17 mouse bone marrow stromal cell layers to derive CD34+ HPCs which were subsequently grown in specific cytokine differentiation media to promote the development of DCs. The hES derived DCs (hES-DC) were subjected to phenotypic and functional analyses and compared with DCs derived from fetal liver CD34+ HPC (FL-DC). The mature hES-DCs displayed typical DC morphology consisting of veiled stellate cells. The hES-DCs also displayed characteristic phenotypic surface markers CD1a, HLA-DR, B7.1, B7.2, and DC-SIGN. The hES-DCs were found to be capable of antigen uptake and stimulating naïve allogeneic CD4+ T cells in a mixed leukocyte reaction assay. Furthermore, the hES-DCs supported productive HIV-1 viral infection akin to standard DCs.ConclusionPhenotypically normal and functionally competent DCs that support HIV-1 infection can be derived from hES cells. hES-DCs can now be exploited in applied immunology and HIV-1 infection studies. Using gene therapy approaches, it is now possible to generate HIV-1 resistant DCs from anti-HIV gene transduced hES-CD34+ hematopoietic progenitor cells.

CsA does not inhibit alloantigen presentation by skin veiled (dendritic) cells

CsA does not inhibit alloantigen presentation by skin veiled (dendritic) cells

Accessory Cells with a Veiled Morphology and Movement Pattern Generated from Monocytes after Avoidance of Plastic Adherence and of NADPH Oxidase Activation. A Comparison with GM-CSF/IL-4-Induced Monocyte-Derived Dendritic Cells

Veiled cells (VC) present in afferent lymph transport antigen from the periphery to the draining lymph nodes. Although VC in lymph form a heterogeneous population, some of the cells clearly belong on morphological grounds to the Langerhans cell (LC)/ dendritic cell (DC) series. Here we show that culturing monocytes for 24 hrs while avoiding plastic adherence (polypropylene tubes) and avoiding the activation of NADPH oxidase (blocking agents) results in the generation of a population of veiled accessory cells. The generated VC were actively moving cells like lymph-borne VC in vivo. The monocyte (mo)-derived VC population existed of CD14(dim/-) and CD14(brighT) cells. Of these the CD14(dim/-) VC were as good in stimulating allogeneic T cell proliferation as immature DC (iDC) obtained after one week of adherent culture of monocytes in granulocyte-macrophage-colony stimulating factor (GM-CSF)/interleukin (IL)-4. This underscores the accessory cell function of the mo-derived CD14(dim/-) VC. Although the CD14(dim/-)VC had a modest expression of the DC-specific marker CD83 and were positive for S100, expression of the DC-specific markers CD1a, Langerin, DC-SIGN, and DC-LAMP were absent. This indicates that the here generated CD14(dim/-) VC can not be considered as classical LC/DC. It was also impossible to turn the CD14(dim/-) mo-derived VC population into typical DC by culture for one week in GM-CSF/IL-4 or LPS. In fact the cells died tinder such circumstances, gaining some macrophage characteristics before dying. The IL-12 production from mo-derived CD14(dim/-) VC was lower, whereas the production of IL-10 was higher as compared to iDC. Consequently the T cells that were stimulated by these mo-derived VC produced less IFN-gamma as compared with T cells stimulated by iDC. Our data indicate that it is possible to rapidly generate a population of CD14(dim/-) veiled accessory cells from monocytes. The marker pattern and cytokine production of these VC indicate that this population is not a classical DC population. The cells might earlier be related to the veiled macrophage-like cells also earlier described in afferent lymph.

An abnormal adherence of monocytes to fibronectin in thyroid autoimmunity has consequences for cell polarization and the development of veiled cells.

Blood monocytes of patients with thyroid autoimmune disease (TAID) display defects in rearranging their cortical actomyosin cytoskeleton ('polarize') in response to chemoattractants. Such rearrangements also take place after the adherence of monocytes to the extracellular matrix (ECM). It is therefore not surprising that monocytes are primed after fibronectin (FN) adherence, displaying an enhanced polarization toward chemoattractants. We investigated the integrin expression and chemoattractant-induced polarization of monocytes of TAID patients before and after FN adherence. Since cytoskeletal rearrangements are also required during the transition of monocytes into veiled antigen-presenting cells (VCs), we investigated such transition of FN-adherent monocytes of TAID patients. Adherent and nonadherent monocyte populations from TAID patients and healthy controls were subjected to a polarization test with the chemoattractant fMLP (or MCP-1), FACS analyses (FITC-labelled FN, CD29, CD49e, d, b and a) and tested for their capability to develop into veiled APC. Monocytes of healthy individuals showed an improved chemoattractant-induced cell polarization after FN adherence, not reflected by TAID monocytes, in which chemoattractant-induced polarization worsened. Monocytes of healthy individuals up-regulated CD49e and d integrins and their capability to bind FITC-labelled FN after adherence to a FN-coated plate, as well as enhancing their capability to generate T cell-stimulatory VCs. Monocytes of TAID patients did not. These data indicate that integrin- (and chemokine-) mediated functions are hampered in monocytes of TAID patients. Because integrin action is pivotal to processes such as monocyte adherence to endothelial cells, uropod formation, migration into tissues and differentiation into APC and macrophages, these defects might underly immune dysbalances important in thyroid autoimmune development.

Cutaneous CsA-resistant veiled (dendritic) cells are responsible for uncontrolled skin allograft rejection

Cutaneous CsA-resistant veiled (dendritic) cells are responsible for uncontrolled skin allograft rejection

Skin allografts—host cutaneous veiled cells initiate rejection reaction by indirect pathway of allorecognition

Skin allografts—host cutaneous veiled cells initiate rejection reaction by indirect pathway of allorecognition

Dendritic cells, implications on function from studies of the afferent lymph veiled cell

Studies of afferent lymph veiled cells (ALVC) show that the full biological function of dendritic cells in peripheral tissue is not explained by a simple model in which immature dendritic cells at the body surface take up antigen, migrate via the afferent lymph ducts, mature and then effectively present antigens to T-cells in the draining lymph node. Furthermore, it is evident from various investigations that the dendritic cells in afferent lymph draining from the body surfaces are not a homogeneous population of cells. They comprise a mixture of cell phenotypes defined by staining with monoclonal antibodies, and the different sub-populations have distinct biological functions and roles in vivo. The molecular basis for differences between the function of afferent lymph dendritic cell subsets is only now being explored and defined but some progress has been made in understanding the role of co-stimulatory molecules. It should be possible to exploit knowledge of the functions of these cells and aid future vaccination strategies in domesticated animals thereby improving animal health and reducing economic loss, and, as a consequence, improving human health. By deliberately targeting functionally distinct subsets of either precursor or mature dendritic cells in vivo, it should become feasible to achieve an appropriately biased immune response.

Sequence analysis or rat integrin alphaE1 and alphaE2 subunits: tissue expression reveals phenotypic similarities between intraepithelial lymphocytes and dendritic cells in lymph.

The integrin alphaOX-62 subunit is defined by the OX-62 monoclonal antibody that was raised against rat dendritic cells in lymph (veiled cells) and shows properties similar to those of human alphaE that is predominantly expressed on intraepithelial lymphocytes. To clone alphaOX-62, rat probes generated using primers specific for the human alphaE sequence were used to screen rat T cell cDNA libraries. cDNA clones encoding two similar but not identical alpha subunits that are closely related but distinct from human alphaE were isolated. alphaE1 is predicted to be the rat homolog of mouse alphaM290 and alphaE2 corresponds to rat alphaOX-62. Immunofluorescence analysis revealed that mouse alphaE1 and rat alphaE2 are expressed in dendritic epidermal T cells in the skin, intraepithelial lymphocytes in the small intestine and in cells with a dendritic morphology present at sites where gammadelta T cells occur in lymphoid organs. Unexpectedly, in veiled cells alphaE2 is co-expressed with intracellular CD3-delta and a 33-kDa CD3 chain but not the T cell receptor. These findings suggest that veiled cells may be derived from a lymphoid precursor. Furthermore, veiled cells show phenotypic similarities to intraepithelial lymphocytes.

Antigen-presenting cells in nasal allergy.

Antigen-presenting cells in nasal allergy.

Dendritic cells in allergy.

Dendritic cells in allergy.

Differentiation of monocytes into dendritic cells in a model of transendothelial trafficking.

Essential to the dendritic cell system of antigen-presenting cells are the veiled dendritic cells that traverse afferent lymph to enter lymph nodes, where they initiate immune responses. The origin of veiled cells, which were discovered 20 years ago, is unclear. Monocytes cultured with endothelium differentiated into dendritic cells within 2 days, particularly after phagocytosing particles in subendothelial collagen. These nascent dendritic cells migrated across the endothelium in the ablumenal-to-lumenal direction, as would occur during entry into lymphatics. Monocytes that remained in the subendothelial matrix became macrophages. Therefore, monocytes have two potential fates associated with distinct patterns of migration.

Identification of two distinct populations of dendritic cells in afferent lymph that vary in their ability to stimulate T cells.

Immunofluorescent staining and flow cytometric analysis of dendritic cells from cattle afferent lymph has established that within the afferent lymph veiled cells (ALVC) there are two phenotypically distinct, major populations. One is CD11a+, CD5+, CD21- and expresses the bovine WC10 (workshop cluster 10) molecule and the Ag recognized by mAb CC81 but is not recognized by mAbs CC149 and IL-A24. The second ALVC subpopulation is CD11a-, CD5-, CD21+/-, workshop cluster 10- and is not recognized by mAb CC81 but is recognized by mAb CC149. Thus, the two populations, which can be identified by staining for CD11a, are defined by the differential expression of a number of Ag. The ALVC populations had differing capacities to stimulate T cells. CD11a- ALVC were more effective at stimulating proliferative responses in allogeneic CD4+ T cells and CD8+ T cells. This was not related to binding of CTLA4Ig or CD40L fusion proteins, implying similar levels of expression of their ligands, CD80 and CD86 or CD40. Both subsets were able to present OVA to resting memory CD4+ T cells, indicating that both were able to take up and process soluble native protein. In contrast, the CD11a- ALVC were more effective in presenting respiratory syncytial virus Ag to resting CD4+ T cells. Considering the central role of dendritic cells in the initiation of immune responses in naive animals, the two cell types may have different roles in the induction of primary responses induced following infection or immunization.

Enrichment and characterization of dendritic cells from rat renal mesangium.

Dendritic cells (DC) initiate primary immune reactions and are distributed throughout most tissues. The most potent DC population of the kidney has long been suggested to reside within the glomerular mesangium. Using LEW.1A rats, we enriched and characterized such low-density cells. Mesangial DC generally exhibited round to oval cell bodies and cytoplasmic veils. Phenotypically, these cells were 100% OX-6++, 45% OX-42++, 35% ED1low, 10% OX-62low, and negative for ED2 and alpha-naphtylbutyrate esterase. Introducing a new monoclonal antibody, R3, which stains a subset of splenic DC, we showed strong antigen expression on 60% of mesangial DC. Correlating cell populations were detected immunohistochemically. Functionally, mesangial DC potently stimulated allogeneic mixed leucocyte reactions, but did not phagocytose opsonized Escherichia coli. In addition to their striking phenotypic similarity with autologous splenic DC, mesangial DC exhibited 88% of the allostimulatory activity of splenic DC. Calculation indicated approximately two mesangial DC per glomerulum. We suggest that these cells comprise different maturation-dependent subsets. The OX-62 integrin especially appears to be expressed only on mature mesangial DC, which may correlate to lymphoid veiled cells or interdigitating DC. An employment of mesangial DC in experimental models of acute allograft rejection or glomerulonephritis is discussed.

The role of skin dendritic cells in the initiation of human immunodeficiency virus infection

Human immunodeficiency virus (HIV) can be transmitted by accidental puncture with needles containing HIV-contaminated blood. However, the molecular and cellular interactions that occur between HIV and cells of the skin following percutaneous needlestick injury are unknown. Direct inoculation of exogenous virus into recipient blood vessels of the dermis is possible. In addition, skin dendritic cells (DC; e.g., epidermal Langerhans cells, dermal DC, lymphatic veiled cells) may also play a role in the initiation of HIV infection. Evidence to suggest that DC are important in primary HIV infection is derived largely from in vitro experiments and animal models. For example, cutaneous DC can be infected with HIV in vitro, can capture HIV on their cell surface (independent from DC infection), and can efficiently transmit HIV to CD4+ T cells. In recent in vivo experiments using rhesus macaques, submucosal DC were the first cells infected following intravaginal exposure to simian immunodeficiency virus (SIV). In this review, I discuss the possible immunologic events that occur within skin and draining lymph nodes following needlestick exposure to HIV-contaminated blood, with a particular emphasis on DC-HIV interactions.

Reactivity of antibodies directed against human antigens with surface markers on canine leukocytes.

A panel of anti-human antibodies was used in order to investigate their cross-reactivity with canine leukocytes. The labeling was carried out at the microscopic level by immunocytochemical staining. Of 50 antibodies 22 cross-reacted with canine leukocytes from afferent lymph and peripheral blood. All leukocytes reacted with MHM23 (CD18). Two anti-HLA DR antibodies, DK22 and L243, reacted mostly with veiled cells and with PHA-stimulated lymphocytes, whereas TAL1B5 was cross-reactive with all canine lymphocytes. The activation markers CD25, Ki-67, PCNA were identified on PHA-stimulated lymphocytes with ACT1, Ki-67 and PC10 clone produced antibodies. Canine eosinophils reacted with MHM6 (CD23) antibody. A large number of antibodies reacted with canine lymph veiled cells. Canine granulocytes and a subset of lymphocytes were stained with the anti-CD15 antibody only after treatment of cytospins with neuraminidase.

Afferent lymph veiled cells stimulate proliferative responses in allogeneic CD4+ and CD8+ T cells but not gamma delta TCR+ T cells.

Dendritic cells were identified in afferent lymph derived by lymphatic cannulation of cattle, stained with monoclonal antibody (mAb) to the bovine workshop cluster 6 (WC6) antigen, which is highly expressed on bovine afferent lymph veiled cells, and sorted with a fluorescence-activated cell sorter. These cells expressed major histocompatibility complex (MHC) class I and II and CD1b but not CD14. They bound human and murine CTLA4-immunoglobulin (CTLA4-Ig) fusion proteins indicating expression of CD80 and or CD86. Dendritic cells induced proliferative responses in allogeneic CD4+ and CD8+ cells sorted from blood but did not induce responses in purified allogeneic WC1+, gamma/delta T cells, which are CD2-, CD4-, CD8- and are the major gamma delta T-cell population in cattle blood, even when interleukin-2 (IL-2) was added to cultures. A WC1-, CD2+ gamma delta T-cell receptor (TCR)+ population predominates in cattle spleens and proliferation of a T-cell line with this phenotype was not induced by allogeneic dendritic cells, with or without added IL-2. The observations imply that the ligand for the gamma delta TCR expressed on the two populations is not present on allogeneic dendritic cells or that the costimulatory molecules expressed on dendritic cells that render them highly effective at stimulating MHC class I- and class II-restricted CD8+ and CD4+ T cells are not recognized by the WC1+ or WC1- gamma/delta T cells. Expression of CD28 by the four cell types was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Purified CD4+ and CD8+ cells both produced CD28 transcripts but neither purified WC1+ cells nor the WC1- gamma delta TCR+ cell line did so. The findings indicate that CD80 and or CD86 are involved in the stimulation of CD4+ and CD8+ alpha beta TCR+ T cells but not in the stimulation of either of the two gamma delta TCR+ populations.

Murine follicular dendritic cells: accessory activities in vitro.

Follicular dendritic cells (FDC) are accessory cells found in association with B lymphocytes in the follicles of stimulated secondary lymphoid tissues. Accessory cells are known to collaborate with lymphocytes and enable them to carry out their essential functions. FDC present antigen (Ag) to B cells and facilitate B cell functions; they may be thought of as B cell-associated accessory cells. In contrast, other dendritic cells, including Langerhans cells, interdigitating cells, and veiled cells, interact primarily with T lymphocytes. These dendritic accessory cells, collectively referred to as dendritic cells (DC) in vitro, present antigen to T cells and may be thought of as T cell-associated accessory cells (TEW et al. 1992a). Data supporting the accessory activities of most accessory cells, such as DC and macrophages, have been collected using in vitro experimentation. In contrast, most of the data available on FDC has been collected using in vivo models. We have reviewed these in vivo data on murine FDC in a number of recent articles.