Low-density lipoprotein associates with extracellular vesicles via apolipoprotein B

Background: We have shown recently that low-density lipoprotein (LDL) co-isolates with extracellular vesicles (EVs) derived from blood plasma and the supernatant of platelet concentrates. Furthermore, we found that with current isolation protocols, EVs and LDL cannot be separated. By transmission el...

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Bibliographic Details
Published inJournal of extracellular vesicles Vol. 7; p. 82
Main Authors Sodar, Barbara W, Pálóczi, Krisztina, Visnovitz, Tamás, Vukman, Krisztina V, Pállinger, Éva, Kovács, Árpád, Tóth, Eszter Á, Hegyesi, Hargita, Kittel, Ágnes, Tóth, Sára, Buzas, Edit
Format Journal Article
LanguageEnglish
Published Abingdon John Wiley & Sons, Inc 01.01.2018
Subjects
Apolipoprotein B
Apolipoprotein E
Apolipoproteins
Centrifugation
Extracellular vesicles
Flow cytometry
Gravity
Latex beads
Lipophilic
Lipoproteins
Low density lipoprotein
Monocytes
Monocytic leukemia
Sulfates
Transmission electron microscopy
Hungary
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Summary:Background: We have shown recently that low-density lipoprotein (LDL) co-isolates with extracellular vesicles (EVs) derived from blood plasma and the supernatant of platelet concentrates. Furthermore, we found that with current isolation protocols, EVs and LDL cannot be separated. By transmission electron microscopy we also demonstrated the association of EVs with LDL in vitro. Methods: We labeled THP-1 human monocytic leukemia cells with the lipophilic dyes PKH67 and DiI. After labeling, small (d < 200 nm) and medium sized (d: ~ 200-800 nm) EVs were isolated by differential centrifugation and gravity-driven filtration from the supernatant. To exclude the possible effect of bovine lipoproteins, we used a 24 h serum free incubation for EV production. Sulfate-aldehyde latex beads were coated with native, oxidized and acetylated LDLs as well as with purified native apolipoproteins (apoA1, apoB, ap°C2 and apoE). After blocking with BSA and glycin, fluorescently labeled EVs were incubated with the beads. Fluorescence of the beads resulting from that of the attached EVs, was analysed by flow cytometry. EV adhesion to different coatings was compared both to the bare and to the blockedonly beads. Results: Both small and medium sized EVs showed significant adhesion to apoB (p < 0.05). There was no difference between the signals of small and medium EVs. We also observed adhesion to native, oxidized and acetylated LDLs, apoA1 and ap°C2. However, in the case of apoE, no binding was detected. Summary/Conclusion: The interaction between LDL and EVs might be mediated by the apolipoprotein B component of LDL.
ISSN:2001-3078