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. 2021 Apr 30;9(4):e002189. doi: 10.1136/jitc-2020-002189

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© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Plasma-derived extracellular vesicles (EVs) contain CD160. (A) Representative flow cytometry plots of CD160 expression in CD8⁺ and CD4⁺ T cells untreated versus treated with the plasma from patients with chronic lymphocytic leukemia (CCL) (using 5%, 10% and 20% plasma) after 72 hours of in vitro culture. (B) Cumulative data of percentages of CD160 expressing cells among CD8⁺, and (C) CD4⁺ T cells either untreated or treated with indicated plasma concentrations after 72 hours. (D) Quantification of EV numbers isolated from the plasma of CLLs versus healthy controls (HCs) by Exocet ELISA kit. (E) ImageStream plots of plasma-derived EVs showing the expression of CD9, CD63 and CD160, bright field (BF). (F) ImageStream plots of plasma-derived EVs showing expression of CD63, CD81 and CD160. (G) Representative western blot (WB) images of plasma-derived EVs from HCs and patients with CLL depicting CD160 presence. (H) Cumulative data showing normalized arbitrary units of CD160/actin in plasma-derived EVs in HCs versus patients with CLL. (I) Representative WB images of plasma-derived EVs depicting CD9 expression, and (J) cumulative data showing normalized arbitrary units of CD9/actin in plasma-derived EVs in HCs versus patients with CLL. (K) Representative WB images of plasma-derived EVs depicting CD63 expression, and (L) cumulative data showing normalized arbitrary units of CD63/GAPDH in plasma-derived EVs in HCs versus patients with CLL. (M) Representative WB images of plasma-derived EVs depicting CD81 expression, and (N) cumulative data showing normalized arbitrary units of CD160/actin in plasma-derived EVs in HCs versus patients with CLL. Actin was used as a loading control to normalize protein amounts of CD81, CD9, and CD160, and GAPDH was used as a loading control to normalize protein amount of CD63. Each dot/band represents data from a subject.