Differential Interaction of Platelet-Derived Extracellular Vesicles with Leukocyte Subsets in Human Whole Blood

Weiss, René; Gröger, Marion; Rauscher, Sabine; Fendl, Birgit; Eichhorn, Tanja; Fischer, Michael B.; Spittler, Andreas; Weber, Viktoria

doi:10.1038/s41598-018-25047-x

Cited by 46 publications

(65 citation statements)

References 66 publications

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“…Samples were further diluted 1:5 in PBS and analyzed on a Cytoflex flow cytometer (Beckman Coulter, USA). For creating a size-based gate <1000 nm for EV detection we used 100, 200, 500 and 1000 nm green fluorescent-labeled silica beads (Kisker Biotech, Germany) as illustrated in Figure 4B [46]- [48]. Based on the separation of fluorescent EVs from particle background we only recorded fluorescence-positive events using double fluorescence triggering in the FITC and APC channels.…”

Section: Ev Identity By Western Blot and Flow Cytometrymentioning

confidence: 99%

A functional corona around extracellular vesicles enhances angiogenesis during skin regeneration and signals in immune cells

Wolf

Poupardin

Ebner-Peking

et al. 2019

Preprint

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words)Allogeneic regenerative cell therapy has shown surprising results despite lack of engraftment of the transplanted cells. Their efficacy was so far considered to be mostly due to secreted trophic factors. We hypothesized that extracellular vesicles (EVs) can also contribute to their mode of action. Here we provide evidence that EVs derived from therapeutic placental-expanded (PLX) stromal cells are potent inducers of angiogenesis and modulate immune cell proliferation in a dose-dependent manner.Crude EVs were enriched >100-fold from large volume PLX conditioned media via tangential flow filtration (TFF) as determined by tunable resistive pulse sensing (TRPS).Additional TFF purification was devised to separate EVs from cell-secreted soluble factors. EV identity was confirmed by western blot, calcein-based flow cytometry and electron microscopy. Surface marker profiling of tetraspanin-positive EVs identified expression of cell-and matrix-interacting adhesion molecules. Differential tandem mass tag proteomics comparing PLX-EVs to PLX-derived soluble factors revealed significant differential enrichment of 258 proteins in purified PLX-EVs involved in angiogenesis, cell movement and immune system signaling. At the functional level, PLX-EVs and cells inhibited T cell mitogenesis. PLX-EVs and soluble factors displayed dose-dependent proangiogenic potential by enhancing tube-like structure formation in vitro.Our findings indicate that the mode of PLX action involves an EV-mediated proangiogenic function and immune response modulation that may help explaining clinical efficacy beyond presence of the transplanted allogeneic cells.

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Section: Ev Identity By Western Blot and Flow Cytometrymentioning

confidence: 99%

A functional corona around extracellular vesicles enhances angiogenesis during skin regeneration and signals in immune cells

Wolf

Poupardin

Ebner-Peking

et al. 2019

Preprint

Self Cite

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“…Interestingly, this distinction depends on the affinity of the binding, which is approximately 100-fold higher for granulocytes with respect to lymphocytes [15,20,21]. Such interactions justify the selective effects of EVs to their target cells.…”

Section: Physiological Ev Effects In Immunologymentioning

confidence: 99%

“…The human blood plasma contains many EVs rich in miRNAs of various types. Among these miRNAs, some are targeted from donors to distinct immunological cells; for example, monocytes and lymphocytes [15][16][17]. miRNAs also appear critical for dendritic cells, with ensuing antipathogenic effects and tolerant phenotypes of T lymphocyte target cells [18][19][20].…”

Section: Physiological Ev Effects In Immunologymentioning

confidence: 99%

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Extracellular vesicles, news about their role in immune cells: physiology, pathology and diseases

Meldolesi

2019

Clinical and Experimental Immunology

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Two types of extracellular vesicles (EVs), exosomes and ectosomes, are generated and released by all cells, including immune cells. The two EVs appear different in many properties: size, mechanism and site of assembly, composition of their membranes and luminal cargoes, sites and processes of release. In functional terms, however, these differences are minor. Moreover, their binding to and effects on target cells appear similar, thus the two types are considered distinct only in a few cases, otherwise they are presented together as EVs. The EV physiology of the various immune cells differs as expected from their differential properties. Some properties, however, are common: EV release, taking place already at rest, is greatly increased upon cell stimulation; extracellular navigation occurs adjacent and at distance from the releasing cells; binding to and uptake by target cells are specific. EVs received from other immune or distinct cells govern many functions in target cells. Immune diseases in which EVs play multiple, often opposite (aggression and protection) effects, are numerous; inflammatory diseases; pathologies of various tissues; and brain diseases, such as multiple sclerosis. EVs also have effects on interactive immune and cancer cells. These effects are often distinct, promoting cytotoxicity or proliferation, the latter together with metastasis and angiogenesis. Diagnoses depend on the identification of EV biomarkers; therapies on various mechanisms such as (1) removal of aggression-inducing EVs; (2) EV manipulations specific for single targets, with insertion of surface peptides or luminal miRNAs; and (3) removal or re-expression of molecules from target cells.

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“…Platelet-derived membrane vesicles are available to carry platelet-derived signals to other cells under physiological conditions (17)(18)(19). Additionally, it has been recognized that the platelets and PMPs affect on the activity of other cells, such as monocytes, neutrophils and endothelial cells (20,21). Unlike PMPs, exosomes do not obtain the surface markers of the activated platelet membranes (22, 23).…”

Section: Introductionmentioning

confidence: 99%

The role of platelet microparticles in the production of antibodies from B lymphocytes against HLA-DR antigen in vitro

Khayati

Yari

2019

IJPHO

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Background: Platelets can activate B cells and stimulate them for the production of antibodies. Since platelet microparticles (PMPs) originate from platelets, they may have the same virtue. In the present study, the effect of PMPs was investigated on the production of human leukocyte antigen (HLA)-specific antibody from B cells in vitro. Materials and Methods: In this experimental study, HLA-DR antigen was solubilized from the immortalized B lymphocytes (Daudi cell line) and purified using the affinity chromatography. Antigen properties were determined by the ELISA technique. PMPs were isolated from platelet concentrate bags by centrifugation. Fresh blood products were prepared from the Innovation Center of Iranian Blood Transfusion Organization (IBTO) and B lymphocytes were purified by the MACS method. B cells were exposed with PMPs and HLA-DR antigens in the culture medium. On the third day of culture, the culture supernatant was examined in terms of antibody production using the ELISA test. The results were analyzed using paired sample T-test and P-value <0.05 was considered statistically significant. Results: The specificity of the purified HLA-DR antigen was confirmed using the anti-HLA-DR antibody and ELISA technique in the presence of appropriate controls. The results showed that PMPs could stimulate the production of antibodies from B cells. The difference between the case and control was significant (P-value=0.001). Although total immunoglobulin (IgG) was higher in HLA-DR-treated wells, HLA-DR-specific antibodies were not identified by ELISA technique. Conclusion: PMPs have the capability to induce IgG antibodies from B cells. In order to ensure the production of specific antibodies, further testing is required with high sensitivity.

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Differential Interaction of Platelet-Derived Extracellular Vesicles with Leukocyte Subsets in Human Whole Blood

Cited by 46 publications

References 66 publications

A functional corona around extracellular vesicles enhances angiogenesis during skin regeneration and signals in immune cells

A functional corona around extracellular vesicles enhances angiogenesis during skin regeneration and signals in immune cells

Extracellular vesicles, news about their role in immune cells: physiology, pathology and diseases

The role of platelet microparticles in the production of antibodies from B lymphocytes against HLA-DR antigen in vitro

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