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  1. Article ; Online: Unveiling

    Bertuccini, Lucia / Boussadia, Zaira / Salzano, Anna Maria / Vanni, Ilaria / Passerò, Ilaria / Nocita, Emanuela / Scaloni, Andrea / Sanchez, Massimo / Sargiacomo, Massimo / Fiani, Maria Luisa / Tosini, Fabio

    Frontiers in cellular and infection microbiology

    2024  Volume 14, Page(s) 1367359

    Abstract: Cryptosporidium ... ...

    Abstract Cryptosporidium parvum
    MeSH term(s) Extracellular Vesicles/metabolism ; Cryptosporidium parvum/metabolism ; Sporozoites/metabolism ; Protozoan Proteins/metabolism ; Protozoan Proteins/analysis ; Microscopy, Electron, Transmission ; Animals ; Cryptosporidiosis/parasitology ; Humans ; Proteome/analysis ; Proteomics ; Flow Cytometry
    Chemical Substances Protozoan Proteins ; Proteome
    Language English
    Publishing date 2024-04-10
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2619676-1
    ISSN 2235-2988 ; 2235-2988
    ISSN (online) 2235-2988
    ISSN 2235-2988
    DOI 10.3389/fcimb.2024.1367359
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Exploiting Manipulated Small Extracellular Vesicles to Subvert Immunosuppression at the Tumor Microenvironment through Mannose Receptor/CD206 Targeting.

    Fiani, Maria Luisa / Barreca, Valeria / Sargiacomo, Massimo / Ferrantelli, Flavia / Manfredi, Francesco / Federico, Maurizio

    International journal of molecular sciences

    2020  Volume 21, Issue 17

    Abstract: Immunosuppression at tumor microenvironment (TME) is one of the major obstacles to be overcome for an effective therapeutic intervention against solid tumors. Tumor-associated macrophages (TAMs) comprise a sub-population that plays multiple pro-tumoral ... ...

    Abstract Immunosuppression at tumor microenvironment (TME) is one of the major obstacles to be overcome for an effective therapeutic intervention against solid tumors. Tumor-associated macrophages (TAMs) comprise a sub-population that plays multiple pro-tumoral roles in tumor development including general immunosuppression, which can be identified in terms of high expression of mannose receptor (MR or CD206). Immunosuppressive TAMs, like other macrophage sub-populations, display functional plasticity that allows them to be re-programmed to inflammatory macrophages. In order to mitigate immunosuppression at the TME, several efforts are ongoing to effectively re-educate pro-tumoral TAMs. Extracellular vesicles (EVs), released by both normal and tumor cells types, are emerging as key mediators of the cell to cell communication and have been shown to have a role in the modulation of immune responses in the TME. Recent studies demonstrated the enrichment of high mannose glycans on the surface of small EVs (sEVs), a subtype of EVs of endosomal origin of 30-150 nm in diameter. This characteristic renders sEVs an ideal tool for the delivery of therapeutic molecules into MR/CD206-expressing TAMs. In this review, we report the most recent literature data highlighting the critical role of TAMs in tumor development, as well as the experimental evidences that has emerged from the biochemical characterization of sEV membranes. In addition, we propose an original way to target immunosuppressive TAMs at the TME by endogenously engineered sEVs for a new therapeutic approach against solid tumors.
    MeSH term(s) Animals ; Extracellular Vesicles/immunology ; Extracellular Vesicles/metabolism ; Extracellular Vesicles/pathology ; Humans ; Immune Tolerance/immunology ; Lectins, C-Type/metabolism ; Macrophages/immunology ; Macrophages/metabolism ; Macrophages/pathology ; Mannose-Binding Lectins/metabolism ; Neoplasms/immunology ; Neoplasms/metabolism ; Neoplasms/pathology ; Receptors, Cell Surface/metabolism ; Tumor Microenvironment/immunology
    Chemical Substances Lectins, C-Type ; Mannose-Binding Lectins ; Receptors, Cell Surface ; mannose receptor
    Language English
    Publishing date 2020-08-31
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms21176318
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Metabolic labelling of a subpopulation of small extracellular vesicles using a fluorescent palmitic acid analogue.

    Barreca, Valeria / Boussadia, Zaira / Polignano, Deborah / Galli, Lorenzo / Tirelli, Valentina / Sanchez, Massimo / Falchi, Mario / Bertuccini, Lucia / Iosi, Francesca / Tatti, Massimo / Sargiacomo, Massimo / Fiani, Maria Luisa

    Journal of extracellular vesicles

    2023  Volume 12, Issue 12, Page(s) e12392

    Abstract: Exosomes are among the most puzzling vehicles of intercellular communication, but several crucial aspects of their biogenesis remain elusive, primarily due to the difficulty in purifying vesicles with similar sizes and densities. Here we report an ... ...

    Abstract Exosomes are among the most puzzling vehicles of intercellular communication, but several crucial aspects of their biogenesis remain elusive, primarily due to the difficulty in purifying vesicles with similar sizes and densities. Here we report an effective methodology for labelling small extracellular vesicles (sEV) using Bodipy FL C16, a fluorescent palmitic acid analogue. In this study, we present compelling evidence that the fluorescent sEV population derived from Bodipy C16-labelled cells represents a discrete subpopulation of small exosomes following an intracellular pathway. Rapid cellular uptake and metabolism of Bodipy C16 resulted in the incorporation of fluorescent phospholipids into intracellular organelles specifically excluding the plasma membrane and ultimately becoming part of the exosomal membrane. Importantly, our fluorescence labelling method facilitated accurate quantification and characterization of exosomes, overcoming the limitations of nonspecific dye incorporation into heterogeneous vesicle populations. The characterization of Bodipy-labelled exosomes reveals their enrichment in tetraspanin markers, particularly CD63 and CD81, and in minor proportion CD9. Moreover, we employed nanoFACS sorting and electron microscopy to confirm the exosomal nature of Bodipy-labelled vesicles. This innovative metabolic labelling approach, based on the fate of a fatty acid, offers new avenues for investigating exosome biogenesis and functional properties in various physiological and pathological contexts.
    MeSH term(s) Extracellular Vesicles/metabolism ; Palmitic Acid/metabolism ; Exosomes/metabolism ; Biological Transport
    Chemical Substances 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene ; Palmitic Acid (2V16EO95H1)
    Language English
    Publishing date 2023-12-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2683797-3
    ISSN 2001-3078 ; 2001-3078
    ISSN (online) 2001-3078
    ISSN 2001-3078
    DOI 10.1002/jev2.12392
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Generation, Quantification, and Tracing of Metabolically Labeled Fluorescent Exosomes.

    Coscia, Carolina / Parolini, Isabella / Sanchez, Massimo / Biffoni, Mauro / Boussadia, Zaira / Zanetti, Cristiana / Fiani, Maria Luisa / Sargiacomo, Massimo

    Methods in molecular biology (Clifton, N.J.)

    2016  Volume 1448, Page(s) 217–235

    Abstract: Over the last 10 years, the constant progression in exosome (Exo)-related studies highlighted the importance of these cell-derived nano-sized vesicles in cell biology and pathophysiology. Functional studies on Exo uptake and intracellular trafficking ... ...

    Abstract Over the last 10 years, the constant progression in exosome (Exo)-related studies highlighted the importance of these cell-derived nano-sized vesicles in cell biology and pathophysiology. Functional studies on Exo uptake and intracellular trafficking require accurate quantification to assess sufficient and/or necessary Exo particles quantum able to elicit measurable effects on target cells. We used commercially available BODIPY(®) fatty acid analogues to label a primary melanoma cell line (Me501) that highly and spontaneously secrete nanovesicles. Upon addition to cell culture, BODIPY fatty acids are rapidly incorporated into major phospholipid classes ultimately producing fluorescent Exo as direct result of biogenesis. Our metabolic labeling protocol produced bright fluorescent Exo that can be examined and quantified with conventional non-customized flow cytometry (FC) instruments by exploiting their fluorescent emission rather than light-scattering detection. Furthermore, our methodology permits the measurement of single Exo-associated fluorescence transfer to cells making quantitative the correlation between Exo uptake and activation of cellular processes. Thus the protocol presented here appears as an appropriate tool to who wants to investigate mechanisms of Exo functions in that it allows for direct and rapid characterization and quantification of fluorescent Exo number, intensity, size, and eventually evaluation of their kinetic of uptake/secretion in target cells.
    Language English
    Publishing date 2016
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-3753-0_16
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer.

    Zanetti, Cristiana / Gallina, Angelo / Fabbri, Alessia / Parisi, Sofia / Palermo, Angela / Fecchi, Katia / Boussadia, Zaira / Carollo, Maria / Falchi, Mario / Pasquini, Luca / Fiani, Maria Luisa / Sargiacomo, Massimo

    International journal of molecular sciences

    2018  Volume 19, Issue 5

    Abstract: In this study, we report how the cholera toxin (CT) A subunit (CTA), the enzyme moiety responsible for signaling alteration in host cells, enters the exosomal pathway, secretes extracellularly, transmits itself to a cell population. The first evidence ... ...

    Abstract In this study, we report how the cholera toxin (CT) A subunit (CTA), the enzyme moiety responsible for signaling alteration in host cells, enters the exosomal pathway, secretes extracellularly, transmits itself to a cell population. The first evidence for long-term transmission of CT's toxic effect via extracellular vesicles was obtained in Chinese hamster ovary (CHO) cells. To follow the CT intracellular route towards exosome secretion, we used a novel strategy for generating metabolically-labeled fluorescent exosomes that can be counted by flow cytometry assay (FACS) and characterized. Our results clearly show the association of CT with exosomes, together with the heat shock protein 90 (HSP90) and Protein Disulfide Isomerase (PDI) molecules, proteins required for translocation of CTA across the ER membrane into the cytoplasm. Confocal microscopy showed direct internalization of CT containing fluorescent exo into CHO cells coupled with morphological changes in the recipient cells that are characteristic of CT action. Moreover, Me665 cells treated with CT-containing exosomes showed an increase in Adenosine 3',5'-Cyclic Monophosphate (cAMP) level, reaching levels comparable to those seen in cells exposed directly to CT. Our results prompt the idea that CT can exploit an exosome-mediated cell communication pathway to extend its pathophysiological action beyond an initial host cell, into a multitude of cells. This finding could have implications for cholera disease pathogenesis and epidemiology.
    MeSH term(s) ADP-Ribosylation Factors/metabolism ; Animals ; CHO Cells ; Cell Line, Tumor ; Cell Membrane/metabolism ; Cholera/etiology ; Cholera Toxin/chemistry ; Cholera Toxin/metabolism ; Cholera Toxin/toxicity ; Cricetinae ; Cricetulus ; Cyclic AMP/metabolism ; Exosomes/metabolism ; HSP90 Heat-Shock Proteins/metabolism ; Humans ; Protein Disulfide-Isomerases/metabolism ; Protein Subunits/metabolism ; Protein Transport
    Chemical Substances HSP90 Heat-Shock Proteins ; Protein Subunits ; Cholera Toxin (9012-63-9) ; Cyclic AMP (E0399OZS9N) ; ADP-Ribosylation Factors (EC 3.6.5.2) ; Protein Disulfide-Isomerases (EC 5.3.4.1)
    Language English
    Publishing date 2018-05-19
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms19051521
    Database MEDical Literature Analysis and Retrieval System OnLINE

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