Extracellular Vesicles from Infected Cells Are Released Prior to Virion Release

Yuriy Kim, Gifty A. Mensah, Sarah Al Sharif, Daniel O. Pinto, Heather Branscome, Sowmya V. Yelamanchili, Maria Cowen, James Erickson, Pooja Khatkar, Renaud Mahieux, Fatah Kashanchi

Research output: Contribution to journalArticlepeer-review

Abstract

Here, we have attempted to address the timing of EV and virion release from virally infected cells. Uninfected (CEM), HIV-1-infected (J1.1), and human T cell leukemia virus-1 (HTLV-1)-infected (HUT102) cells were synchronized in G0. Viral latency was reversed by increasing gene expression with the addition of serum-rich media and inducers. Supernatants and cell pellets were collected post-induction at different timepoints and assayed for extracellular vesicle (EV) and autophagy markers; and for viral proteins and RNAs. Tetraspanins and autophagy-related proteins were found to be differentially secreted in HIV-1- and HTLV-1-infected cells when compared with uninfected controls. HIV-1 proteins were present at 6 h and their production increased up to 24 h. HTLV-1 proteins peaked at 6 h and plateaued. HIV-1 and HTLV-1 RNA production correlated with viral protein expression. Nanoparticle tracking analysis (NTA) showed increase of EV concentration over time in both uninfected and infected samples. Finally, the HIV-1 supernatant from the 6-h samples was found not to be infectious; however, the virus from the 24-h samples was successfully rescued and infectious. Overall, our data indicate that EV release may occur prior to viral release from infected cells, thereby implicating a potentially significant effect of EVs on uninfected recipient cells prior to subsequent viral infection and spread.

Original languageEnglish (US)
JournalCells
Volume10
Issue number4
DOIs
StatePublished - Apr 1 2021

Keywords

  • cART
  • exosomes
  • extracellular vesicles
  • HIV-1
  • HTLV-1

ASJC Scopus subject areas

  • Medicine(all)

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