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Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity.

Publication ,  Journal Article
Ouyang, Y; Bayer, A; Chu, T; Tyurin, VA; Kagan, VE; Morelli, AE; Coyne, CB; Sadovsky, Y
Published in: Placenta
November 2016

INTRODUCTION: Primary human trophoblasts release a repertoire of extracellular vesicles (EVs). Among them are nano-sized exosomes, which we found to suppress the replication of a wide range of diverse viruses. These exosomes contain trophoblastic microRNAs (miRNAs) that are expressed from the chromosome 19 miRNA cluster and exhibit antiviral properties. Here, we report our investigation of the cargo of placental EVs, focusing on the composition and the antiviral properties of exosomes, microvesicles, and apoptotic blebs. METHODS: We isolated EVs using ultracentrifugation and defined their purity using immunoblotting, electron microscopy, and nanoparticle tracking. We used liquid chromatography-electrospray ionization-mass spectrometry, protein mass spectrometry, and miRNA TaqMan card PCR to examine the phospholipids, proteins, and miRNA cargo of trophoblastic EVs and an in vitro viral infection assay to assess the antiviral properties of EVs. RESULTS: We found that all three EV types contain a comparable repertoire of miRNA. Interestingly, trophoblastic exosomes harbor a protein and phospholipid profile that is distinct from that of microvesicles or apoptotic blebs. Functionally, trophoblastic exosomes exhibit the highest antiviral activity among the EVs. Consistently, plasma exosomes derived from pregnant women recapitulate the antiviral effect of trophoblastic exosomes derived from in vitro cultures of primary human trophoblasts. DISCUSSION: When compared to other trophoblastic EVs, exosomes exhibit a unique repertoire of proteins and phospholipids, but not miRNAs, and a potent viral activity. Our work suggests that human trophoblastic EVs may play a key role in maternal-placental-fetal communication.

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Published In

Placenta

DOI

EISSN

1532-3102

Publication Date

November 2016

Volume

47

Start / End Page

86 / 95

Location

Netherlands

Related Subject Headings

  • Trophoblasts
  • Pregnancy
  • Placenta
  • Phospholipids
  • Obstetrics & Reproductive Medicine
  • MicroRNAs
  • Mass Spectrometry
  • Humans
  • Female
  • Extracellular Vesicles
 

Citation

APA
Chicago
ICMJE
MLA
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Ouyang, Y., Bayer, A., Chu, T., Tyurin, V. A., Kagan, V. E., Morelli, A. E., … Sadovsky, Y. (2016). Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta, 47, 86–95. https://doi.org/10.1016/j.placenta.2016.09.008
Ouyang, Yingshi, Avraham Bayer, Tianjiao Chu, Vladimir A. Tyurin, Valerian E. Kagan, Adrian E. Morelli, Carolyn B. Coyne, and Yoel Sadovsky. “Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity.Placenta 47 (November 2016): 86–95. https://doi.org/10.1016/j.placenta.2016.09.008.
Ouyang Y, Bayer A, Chu T, Tyurin VA, Kagan VE, Morelli AE, et al. Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta. 2016 Nov;47:86–95.
Ouyang, Yingshi, et al. “Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity.Placenta, vol. 47, Nov. 2016, pp. 86–95. Pubmed, doi:10.1016/j.placenta.2016.09.008.
Ouyang Y, Bayer A, Chu T, Tyurin VA, Kagan VE, Morelli AE, Coyne CB, Sadovsky Y. Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta. 2016 Nov;47:86–95.
Journal cover image

Published In

Placenta

DOI

EISSN

1532-3102

Publication Date

November 2016

Volume

47

Start / End Page

86 / 95

Location

Netherlands

Related Subject Headings

  • Trophoblasts
  • Pregnancy
  • Placenta
  • Phospholipids
  • Obstetrics & Reproductive Medicine
  • MicroRNAs
  • Mass Spectrometry
  • Humans
  • Female
  • Extracellular Vesicles