Increasing the immune activity of exosomes: the effect of miRNA-depleted exosome proteins on activating dendritic cell/cytokine-induced killer cells against pancreatic cancer

J Zhejiang Univ Sci B. 2016 May;17(5):352-60. doi: 10.1631/jzus.B1500305.

Abstract

Background: Tumor-derived exosomes were considered to be potential candidates for tumor vaccines because they are abundant in immune-regulating proteins, whereas tumor exosomal miRNAs may induce immune tolerance, thereby having an opposite immune function.

Objective: This study was designed to separate exosomal protein and depleted exosomal microRNAs (miRNAs), increasing the immune activity of exosomes for activating dendritic cell/cytokine-induced killer cells (DC/CIKs) against pancreatic cancer (PC).

Methods: PC-derived exosomes (PEs) were extracted from cultured PANC-1 cell supernatants and then ruptured; this was followed by ultrafiltered exosome lysates (UELs). DCs were stimulated with lipopolysaccharide (LPS), PE, and UEL, followed by co-culture with CIKs. The anti-tumor effects of DC/CIKs against PC were evaluated by proliferation and killing rates, tumor necrosis factor-α (TNF-α) and perforin secretion. Exosomal miRNAs were depleted after lysis and ultrafiltration, while 128 proteins were retained, including several immune-activating proteins.

Results: UEL-stimulated DC/CIKs showed a higher killing rate than LPS- and PE-stimulated DC/CIKs.

Conclusions: miRNA-depleted exosome proteins may be promising agonists for specifically activating DC/CIKs against PC.

Keywords: Dendritic cell; Exosome; MicroRNAs; Pancreatic cancer.

MeSH terms

  • Cell Line, Tumor
  • Cell Proliferation
  • Chromatography, Liquid
  • Cytokine-Induced Killer Cells / cytology
  • Cytokine-Induced Killer Cells / immunology*
  • Cytokines / immunology*
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Exosomes / immunology*
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Immune System
  • Lipopolysaccharides
  • MicroRNAs / metabolism*
  • Pancreatic Neoplasms / immunology
  • Pancreatic Neoplasms / metabolism*
  • Proteomics
  • Spectrometry, Mass, Electrospray Ionization
  • Tumor Necrosis Factor-alpha / immunology

Substances

  • Cytokines
  • Lipopolysaccharides
  • MicroRNAs
  • Tumor Necrosis Factor-alpha