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. 2018 May 8;9:956. doi: 10.3389/fimmu.2018.00956

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Copyright © 2018 Danesh, Inglis, Abdel-Mohsen, Deng, Adelman, Schechtman, Heitman, Vilardi, Shah, Keating, Cohen, Jacobs, Pillai, Lacroix, Spinella and Norris.

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 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.

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Cytokine secretion by monocytes stimulated with subtypes of extracellular vesicles (EVs). Monocytes from peripheral blood mononuclear cells of six healthy donors were purified by negative selection. Six replicates of red blood cell-, platelet-, monocyte-, endothelial-, and granulocyte-EVs were prepared as described in the “Materials and Methods” section. Monocytes were cultured unstimulated or incubated with noted EV subtypes for 24 h. (A) Two independent experiments were run with small (enriched for exosomes) and large (enriched for MVs) fractions of granulocyte- and platelet-EVs, and the percentage of monocytes that produced TNF-α was measured by intracellular staining. Representative data showing intracellular cytokine staining of monocytes incubated with small and large fractions of granulocyte- and platelet-EVs. (B) Supernatants were collected at 24 h and were tested using a multiplex cytokine assay for 12 cytokines. Data were analyzed by ANOVA, and each condition was compared with the control condition using a Dunnett’s post-test. Data are shown for 6 of the 12 cytokines tested. (C) The log₁₀ ratio of cytokines induced by incubating monocytes with five subtypes of EVs over the control condition is summarized in a heat map for all 12 cytokines (*p < 0.05, **p < 0.01, and ***p < 0.001).

Cytokine secretion by monocytes stimulated with subtypes of extracellular vesicles (EVs). Monocytes from peripheral blood mononuclear cells of six healthy donors were purified by negative selection. Six replicates of red blood cell-, platelet-, monocyte-, endothelial-, and granulocyte-EVs were prepared as described in the “Materials and Methods” section. Monocytes were cultured unstimulated or incubated with noted EV subtypes for 24 h. (A) Two independent experiments were run with small (enriched for exosomes) and large (enriched for MVs) fractions of granulocyte- and platelet-EVs, and the percentage of monocytes that produced TNF-α was measured by intracellular staining. Representative data showing intracellular cytokine staining of monocytes incubated with small and large fractions of granulocyte- and platelet-EVs. (B) Supernatants were collected at 24 h and were tested using a multiplex cytokine assay for 12 cytokines. Data were analyzed by ANOVA, and each condition was compared with the control condition using a Dunnett’s post-test. Data are shown for 6 of the 12 cytokines tested. (C) The log₁₀ ratio of cytokines induced by incubating monocytes with five subtypes of EVs over the control condition is summarized in a heat map for all 12 cytokines (*p < 0.05, **p < 0.01, and ***p < 0.001).