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Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages

Extracellular vesicles (EVs) are produced by across almost all the living kingdoms and play a crucial role in cell-cell communication processes. EVs are especially important for pathogens, as Plasmodium falciparum (Pf) parasite, the leading causing species in human malaria. Malaria parasites are abl...

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Autores principales: Alfandari, Daniel, Ben Ami Pilo, Hila, Abou Karam, Paula, Dagan, Osnat, Joubran, Carine, Rotkopf, Ron, Regev-Rudzki, Neta, Porat, Ziv
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8825493/
https://www.ncbi.nlm.nih.gov/pubmed/35155269
http://dx.doi.org/10.3389/fcimb.2021.739628
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author Alfandari, Daniel
Ben Ami Pilo, Hila
Abou Karam, Paula
Dagan, Osnat
Joubran, Carine
Rotkopf, Ron
Regev-Rudzki, Neta
Porat, Ziv
author_facet Alfandari, Daniel
Ben Ami Pilo, Hila
Abou Karam, Paula
Dagan, Osnat
Joubran, Carine
Rotkopf, Ron
Regev-Rudzki, Neta
Porat, Ziv
author_sort Alfandari, Daniel
collection PubMed
description Extracellular vesicles (EVs) are produced by across almost all the living kingdoms and play a crucial role in cell-cell communication processes. EVs are especially important for pathogens, as Plasmodium falciparum (Pf) parasite, the leading causing species in human malaria. Malaria parasites are able to modulate the host immune response from a distance via delivering diverse cargo components inside the EVs, such as proteins and nucleic acids. We have previously shown that imaging flow cytometry (IFC) can be effectively used to monitor the uptake of different cargo components of malaria derived EVs by host human monocytes. Here, we take this approach one step further and demonstrate that we can directly investigate the dynamics of the cargo distribution pattern over time by monitoring its distribution within two different recipient cells of the immune system, monocytes vs macrophages. By staining the RNA cargo of the vesicles and monitor the signal we were able to evaluate the kinetics of its delivery and measure different parameters of the cargo’s distribution post internalization. Interestingly, we found that while the level of the EV uptake is similar, the pattern of the signal for RNA cargo distribution is significantly different between these two recipient immune cells. Our results demonstrate that this method can be applied to study the distribution dynamics of the vesicle cargo post uptake to different types of cells. This can benefit significantly to our understanding of the fate of cargo components post vesicle internalization in the complex interface between pathogen-derived vesicles and their host recipient cells.
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spelling pubmed-88254932022-02-10 Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages Alfandari, Daniel Ben Ami Pilo, Hila Abou Karam, Paula Dagan, Osnat Joubran, Carine Rotkopf, Ron Regev-Rudzki, Neta Porat, Ziv Front Cell Infect Microbiol Cellular and Infection Microbiology Extracellular vesicles (EVs) are produced by across almost all the living kingdoms and play a crucial role in cell-cell communication processes. EVs are especially important for pathogens, as Plasmodium falciparum (Pf) parasite, the leading causing species in human malaria. Malaria parasites are able to modulate the host immune response from a distance via delivering diverse cargo components inside the EVs, such as proteins and nucleic acids. We have previously shown that imaging flow cytometry (IFC) can be effectively used to monitor the uptake of different cargo components of malaria derived EVs by host human monocytes. Here, we take this approach one step further and demonstrate that we can directly investigate the dynamics of the cargo distribution pattern over time by monitoring its distribution within two different recipient cells of the immune system, monocytes vs macrophages. By staining the RNA cargo of the vesicles and monitor the signal we were able to evaluate the kinetics of its delivery and measure different parameters of the cargo’s distribution post internalization. Interestingly, we found that while the level of the EV uptake is similar, the pattern of the signal for RNA cargo distribution is significantly different between these two recipient immune cells. Our results demonstrate that this method can be applied to study the distribution dynamics of the vesicle cargo post uptake to different types of cells. This can benefit significantly to our understanding of the fate of cargo components post vesicle internalization in the complex interface between pathogen-derived vesicles and their host recipient cells. Frontiers Media S.A. 2022-01-26 /pmc/articles/PMC8825493/ /pubmed/35155269 http://dx.doi.org/10.3389/fcimb.2021.739628 Text en Copyright © 2022 Alfandari, Ben Ami Pilo, Abou Karam, Dagan, Joubran, Rotkopf, Regev-Rudzki and Porat https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Alfandari, Daniel
Ben Ami Pilo, Hila
Abou Karam, Paula
Dagan, Osnat
Joubran, Carine
Rotkopf, Ron
Regev-Rudzki, Neta
Porat, Ziv
Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title_full Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title_fullStr Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title_full_unstemmed Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title_short Monitoring Distribution Dynamics of EV RNA Cargo Within Recipient Monocytes and Macrophages
title_sort monitoring distribution dynamics of ev rna cargo within recipient monocytes and macrophages
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8825493/
https://www.ncbi.nlm.nih.gov/pubmed/35155269
http://dx.doi.org/10.3389/fcimb.2021.739628
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