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. 2017 May 2;7:1342. doi: 10.1038/s41598-017-01386-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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B cell and monocyte activation and EVs. (A) Plasma EVs (n = 54 EV donors) from young, middle-aged and older individuals were incubated with a different PBMC donor for 24 hrs and analyzed by FACS. (B) cells (CD19⁺) and monocytes (CD11b⁺CD14⁺CD15⁻) were stained with activation markers CD80, CD25 and MHC-II. B cell percentage that express CD25 or CD80 and were positive for EV uptake (CD19⁺PKH⁺CD25⁺ or CD19⁺PKH⁺CD80⁺) were compared to the B cell percentage that express CD25 or CD80 but were not positive for EV uptake (CD19⁺PKH⁻CD25⁺ or CD19⁺PKH⁻CD80⁺). (B) Monocytes that express MHC-II or CD80 and interacted with EVs (CD11b⁺PKH⁺MHC-II⁺ or CD11b⁺PKH⁺CD80⁺) was compared to the monocyte percentage that express MHC-II but did not interact with EVs (CD11b⁺PKH⁻MHC-II⁺ or CD11b⁺PKH⁻CD80⁺). (C) PBMCs from above were treated with LPS or PBS for 2 hrs then EVs (n = 23 a subset of EV donors from A-B) were added for 24 hrs followed by FACS. B cell percentage is shown for (CD19⁺PKH⁺) and monocyte percentage for (CD11b⁺PKH⁺). The histograms are the value from 54 EV samples for A-B and 23 EV samples for C predicted from linear mixed model regression accounting for matching + 95% confidence interval. ***P < 0.001 and **P < 0.01.