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. 2022 Jul 11;29:52. doi: 10.1186/s12929-022-00832-z

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — COVID-19 EVs are derived from activated platelets and induce NET formation via CLEC5A. a EVs from healthy controls (HCs-EVs, n = 2) and COVID-19 patients (COVID19-EVs, n = 5) were harvested by ultracentrifugation, then lysed in RIPA solution before subjected to mass spectrometry analysis. Proteins expressed in COVID-19 EVs, but not in HCs EVs, were further analyzed using the QIAGEN Ingenuity Pathway Analysis (QIAGEN IPA) software. Proteins which were expressed in all the COVID19-EVs were displayed. b, c HCs-EVs (n = 3) and COVID19-EVs (n = 10) were analyzed by flow cytometry, and markers highly activated in COVID-19 platelets were expressed as a heat map (b) or by mean fluorescence intensity (c). d Neutrophils were pre-incubated with anti-CLEC5A mAb (3E12A2, 10 μg/ml), anti-TLR2 mAb (# MAB2616, 10 μg/ml), or both anti-CLEC5A mAb (3E12A2, 10 μg/ml) and anti-TLR2 mAb (# MAB2616, 10 μg/ml), for 30 min at room temperature, followed by incubation with EVs (1 μg/ml) from COVID-19 patients (n = 6) at 37 °C for 3 h. Data are mean ± sd and repeats of at least three independent experiments. n.s.: no significant differences, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (Student’s t-test). N.D: not detectable