Label-Free Proteomic Analysis of Exosomes Secreted from THP-1-Derived Macrophages Treated with IFN-α Identifies Antiviral Proteins Enriched in Exosomes

J Proteome Res. 2019 Mar 1;18(3):855-864. doi: 10.1021/acs.jproteome.8b00514. Epub 2019 Jan 3.


Exosomes are extracellular vesicles that function in intercellular communication. We have previously reported that exosomes play an important role in the transmission of antiviral molecules during interferon-α (IFN-α) treatment. In this study, the protein profiles of THP-1-derived macrophages with or without interferon-α treatment and the exosomes secreted from these cells were analyzed by label-free liquid chromatography-tandem mass spectrometry quantitation technologies. A total of 1845 and 1550 protein groups were identified in the THP-1 macrophages and the corresponding exosomes, respectively. Treating the cells with IFN-α resulted in the differential abundance of 94 proteins in cells and 67 proteins in exosomes (greater than 2.0-fold), among which 23 proteins were up-regulated in both the IFN-α treated cells and corresponding exosomes, while 14 proteins were specifically up-regulated in exosomes but not in the donor cells. GO and KEGG analysis of the identified proteins suggested that IFN-α promoted the abundance of proteins involved in the "defense response to virus" and "type I interferon signaling pathway" in both exosomes and cells. Functional analysis further indicated that exosomes from IFN-α-treated cells exhibited potent antiviral activity that restored the impaired antiviral response of IFN-α in hepatitis B virus-replicating hepatocytes. These results have deepened the understanding of the exosome-mediated transfer of IFN-α-induced antiviral molecules and may provide a new basis for therapeutic strategies to control viral infection.

Keywords: IFN-α; ISGs; exosomes; label-free quantification; macrophages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / analysis
  • Antiviral Agents / metabolism
  • Exosomes / chemistry*
  • Exosomes / metabolism
  • Hepatitis B virus / immunology
  • Humans
  • Immunity, Innate* / drug effects
  • Interferon-alpha / pharmacology*
  • Macrophages / chemistry
  • Macrophages / drug effects
  • Macrophages / metabolism*
  • Proteomics / methods*
  • THP-1 Cells


  • Antiviral Agents
  • Interferon-alpha