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Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer

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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983816/
https://www.ncbi.nlm.nih.gov/pubmed/29783743
http://dx.doi.org/10.3390/ijms19051521
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author 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
author_facet 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
author_sort Zanetti, Cristiana
collection PubMed
description 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.
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spelling pubmed-59838162018-06-05 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 Int J Mol Sci Article 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. MDPI 2018-05-19 /pmc/articles/PMC5983816/ /pubmed/29783743 http://dx.doi.org/10.3390/ijms19051521 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
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
Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title_full Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title_fullStr Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title_full_unstemmed Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title_short Cell Propagation of Cholera Toxin CTA ADP-Ribosylating Factor by Exosome Mediated Transfer
title_sort cell propagation of cholera toxin cta adp-ribosylating factor by exosome mediated transfer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983816/
https://www.ncbi.nlm.nih.gov/pubmed/29783743
http://dx.doi.org/10.3390/ijms19051521
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