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      Nanovesicles released by OKT3 hybridoma express fully active antibodies

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          Abstract

          Recent findings have shown that nanovesicles preparations from either primary immune cells culture supernatants or plasma contain immunoglobulins, suggesting that a natural way of antibody production may be through exosome release. To verify this hypothesis, we used the OKT3 hybridoma clone, which produces a murine IgG2a monoclonal antibody used to reduce rejection in patients undergoing organ transplantation. We showed exosome-associated immunoglobulins in hybridoma supernatants, by Western blot, nanoscale flow cytometry and immunocapture-based ELISA. The OKT3-exo was also being able to trigger cytokines production in both CD4 and CD8 T cells. These results show that nanovesicles contain immunoglobulin and could be used for immunotherapy. These data could lead to a new approach to improve the effectiveness of therapeutic antibodies by exploiting their natural property to be expressed on nanovesicle membrane, that probably render them more stable and as a consequence more capable to interact with their specific ligand in the best way.

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          Most cited references64

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          Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

          ABSTRACT The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.
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            Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles.

            In the 1980s, exosomes were described as vesicles of endosomal origin secreted from reticulocytes. Interest increased around these extracellular vesicles, as they appeared to participate in several cellular processes. Exosomes bear proteins, lipids, and RNAs, mediating intercellular communication between different cell types in the body, and thus affecting normal and pathological conditions. Only recently, scientists acknowledged the difficulty of separating exosomes from other types of extracellular vesicles, which precludes a clear attribution of a particular function to the different types of secreted vesicles. To shed light into this complex but expanding field of science, this review focuses on the definition of exosomes and other secreted extracellular vesicles. Their biogenesis, their secretion, and their subsequent fate are discussed, as their functions rely on these important processes.
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              Exosomes: composition, biogenesis and function

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                Author and article information

                Journal
                J Enzyme Inhib Med Chem
                J Enzyme Inhib Med Chem
                Journal of Enzyme Inhibition and Medicinal Chemistry
                Taylor & Francis
                1475-6366
                1475-6374
                6 January 2021
                2021
                : 36
                : 1
                : 175-182
                Affiliations
                [a ]Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità , Rome, Italy
                [b ]Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità , Rome, Italy
                [c ]FARVA – National Centre for Drug Research and Evaluation, Istituto Superiore di Sanità , Rome, Italy
                [d ]Neuroimmunology Unit, IRCCS Santa Lucia Foundation , Rome, Italy
                [e ]National AIDS Center, Istituto Superiore di Sanità , Rome, Italy
                Author notes

                Supplemental data for this article can be accessed here .

                [*]

                These authors contributed equally to this work.

                CONTACT Stefano Fais stefano.fais@ 123456iss.it Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità , Viale Regina Elena 299, Rome, 00161, Italy
                Article
                1852401
                10.1080/14756366.2020.1852401
                7801098
                33404266
                c6daa895-71c3-4ce7-a18e-617998fd3d57
                © 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Page count
                Figures: 5, Tables: 0, Pages: 8, Words: 5974
                Categories
                Research Article
                Research Paper

                Pharmaceutical chemistry
                exosomes,extracellular vesicles,immunoglobulins,okt3 hybridoma cell line

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