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Lymphatic transport of exosomes as a rapid route of information dissemination to the lymph node.

Srinivasan S, Vannberg FO, Dixon JB - Sci Rep (2016)

Bottom Line: Furthermore, we have demonstrated a differential distribution of exosomes in the draining lymph nodes that is dependent on the lymphatic flow.Lastly, through endpoint analysis of cellular distribution of exosomes in the node, we identified macrophages and B-cells as key players in exosome uptake.Together these results suggest that exosome transfer by lymphatic flow from the periphery to the lymph node could provide a mechanism for rapid exchange of infection-specific information that precedes the arrival of migrating cells, thus priming the node for a more effective immune response.

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Georgia Institute of Technology, Atlanta, GA, USA.

ABSTRACT
It is well documented that cells secrete exosomes, which can transfer biomolecules that impact recipient cells' functionality in a variety of physiologic and disease processes. The role of lymphatic drainage and transport of exosomes is as yet unknown, although the lymphatics play critical roles in immunity and exosomes are in the ideal size-range for lymphatic transport. Through in vivo near-infrared (NIR) imaging we have shown that exosomes are rapidly transported within minutes from the periphery to the lymph node by lymphatics. Using an in vitro model of lymphatic uptake, we have shown that lymphatic endothelial cells actively enhanced lymphatic uptake and transport of exosomes to the luminal side of the vessel. Furthermore, we have demonstrated a differential distribution of exosomes in the draining lymph nodes that is dependent on the lymphatic flow. Lastly, through endpoint analysis of cellular distribution of exosomes in the node, we identified macrophages and B-cells as key players in exosome uptake. Together these results suggest that exosome transfer by lymphatic flow from the periphery to the lymph node could provide a mechanism for rapid exchange of infection-specific information that precedes the arrival of migrating cells, thus priming the node for a more effective immune response.

No MeSH data available.


Characterization of exosomes and beads.(a) Size distribution of HEY exosomes as compared to that of beads. (b) Scanning electron micrograph of exosomes. Scale bar = 500 nm. (c) Expression of CD63 and (d) CD81 on exosomes and beads. (e) Quantitation of CD63 and CD81 on exosomes and beads by flow cytometry (p-value < 0.01).
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f1: Characterization of exosomes and beads.(a) Size distribution of HEY exosomes as compared to that of beads. (b) Scanning electron micrograph of exosomes. Scale bar = 500 nm. (c) Expression of CD63 and (d) CD81 on exosomes and beads. (e) Quantitation of CD63 and CD81 on exosomes and beads by flow cytometry (p-value < 0.01).

Mentions: Exosomes from the HEY cell line were isolated and characterized along with size and density matched polystyrene beads using dynamic light scattering, scanning electron microscopy and for surface marker expression using flow cytometry. The average size of HEY exosomes was 78.82 ± 19.17SD nm as compared to the beads which had an average size of 67.34 ± 13.7SD nm (Fig. 1a). Exosomes had a spherical shape with a diameter of ~60–75 nm as seen from scanning electron microscopy which agreed with previous reports of exosome shape and size reported in literature20 (Fig. 1b). The classical tetraspanin surface markers CD63 and CD81, which are known to be enriched on exosomal membranes4, had an ~80% expression level on HEY exosomes [Fig. 1c–e]. Thus, the HEY exosomes used in this study conformed to known exosomal size ranges, expressed the classical tetraspanin markers and were spherical in shape as previously reported.


Lymphatic transport of exosomes as a rapid route of information dissemination to the lymph node.

Srinivasan S, Vannberg FO, Dixon JB - Sci Rep (2016)

Characterization of exosomes and beads.(a) Size distribution of HEY exosomes as compared to that of beads. (b) Scanning electron micrograph of exosomes. Scale bar = 500 nm. (c) Expression of CD63 and (d) CD81 on exosomes and beads. (e) Quantitation of CD63 and CD81 on exosomes and beads by flow cytometry (p-value < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834495&req=5

f1: Characterization of exosomes and beads.(a) Size distribution of HEY exosomes as compared to that of beads. (b) Scanning electron micrograph of exosomes. Scale bar = 500 nm. (c) Expression of CD63 and (d) CD81 on exosomes and beads. (e) Quantitation of CD63 and CD81 on exosomes and beads by flow cytometry (p-value < 0.01).
Mentions: Exosomes from the HEY cell line were isolated and characterized along with size and density matched polystyrene beads using dynamic light scattering, scanning electron microscopy and for surface marker expression using flow cytometry. The average size of HEY exosomes was 78.82 ± 19.17SD nm as compared to the beads which had an average size of 67.34 ± 13.7SD nm (Fig. 1a). Exosomes had a spherical shape with a diameter of ~60–75 nm as seen from scanning electron microscopy which agreed with previous reports of exosome shape and size reported in literature20 (Fig. 1b). The classical tetraspanin surface markers CD63 and CD81, which are known to be enriched on exosomal membranes4, had an ~80% expression level on HEY exosomes [Fig. 1c–e]. Thus, the HEY exosomes used in this study conformed to known exosomal size ranges, expressed the classical tetraspanin markers and were spherical in shape as previously reported.

Bottom Line: Furthermore, we have demonstrated a differential distribution of exosomes in the draining lymph nodes that is dependent on the lymphatic flow.Lastly, through endpoint analysis of cellular distribution of exosomes in the node, we identified macrophages and B-cells as key players in exosome uptake.Together these results suggest that exosome transfer by lymphatic flow from the periphery to the lymph node could provide a mechanism for rapid exchange of infection-specific information that precedes the arrival of migrating cells, thus priming the node for a more effective immune response.

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Georgia Institute of Technology, Atlanta, GA, USA.

ABSTRACT
It is well documented that cells secrete exosomes, which can transfer biomolecules that impact recipient cells' functionality in a variety of physiologic and disease processes. The role of lymphatic drainage and transport of exosomes is as yet unknown, although the lymphatics play critical roles in immunity and exosomes are in the ideal size-range for lymphatic transport. Through in vivo near-infrared (NIR) imaging we have shown that exosomes are rapidly transported within minutes from the periphery to the lymph node by lymphatics. Using an in vitro model of lymphatic uptake, we have shown that lymphatic endothelial cells actively enhanced lymphatic uptake and transport of exosomes to the luminal side of the vessel. Furthermore, we have demonstrated a differential distribution of exosomes in the draining lymph nodes that is dependent on the lymphatic flow. Lastly, through endpoint analysis of cellular distribution of exosomes in the node, we identified macrophages and B-cells as key players in exosome uptake. Together these results suggest that exosome transfer by lymphatic flow from the periphery to the lymph node could provide a mechanism for rapid exchange of infection-specific information that precedes the arrival of migrating cells, thus priming the node for a more effective immune response.

No MeSH data available.