Inhibition of HBV Replication in HepG2.2.15 Cells by Human Peripheral Blood Mononuclear Cell-Derived Dendritic Cells

Ann Clin Lab Sci. 2015 Fall;45(5):495-501.

Abstract

Anti-HBV therapy is essential for patients awaiting liver transplantation. This study aimed to explore the effects of dendritic cells (DCs) derived from the peripheral blood of hepatitis B patients on the replication of HBV in vivo and to evaluate the biosafety of DCs in clinical therapy. Peripheral blood mononuclear cells (PBMCs) were isolated from HBV-infected patients and maturation-promoting factors and both HBsAg and HBcAg were used to induce DC maturation. Mature DCs and lymphocytes were co-cultured with human hepatocyte cell HL-7702 or HBV-producing human hepatocellular carcinoma cell HepG2.2.15. We found that mature lymphocytes exposed to DCs in vitro did not influence morphology or activities of HL-7702 and HepG2.2.15 cells. Liver function indexes and endotoxin levels in the cell supernatants did not change in these co-cultures. Additionally, supernatant and intracellular HBV DNA levels were reduced when HepG2.2.15 cells were co-cultured with mature lymphocytes that had been cultured with DCs, and HBV covalently closed circular DNA (cccDNA) levels in HepG2.2.15 cells also decreased. Importantly, DC-mediated immunotherapy had no mutagenic effect on HBV genomic DNA by gene sequencing of the P, S, X, and C regions of HBV genomic DNA. We conclude that PBMC-derived DCs from HBV-infected patients act on autologous lymphocytes to suppress HBV replication and these DC clusters showed favorable biosafety.

Keywords: HBV genome sequence; HepG2.2.15; biosafety; dendritic cells; hepatitis B virus.

MeSH terms

  • Cell- and Tissue-Based Therapy / methods
  • Coculture Techniques
  • Culture Media, Conditioned / analysis
  • DNA, Viral / analysis
  • DNA, Viral / genetics
  • Dendritic Cells / physiology*
  • Dendritic Cells / virology
  • Endotoxins / analysis
  • Hep G2 Cells / virology
  • Hepatitis B / blood
  • Hepatitis B / therapy
  • Hepatitis B virus / pathogenicity
  • Hepatitis B virus / physiology*
  • Hepatocytes / physiology
  • Humans
  • Leukocytes, Mononuclear / virology*
  • Lymphocyte Activation
  • Virus Replication / physiology*

Substances

  • Culture Media, Conditioned
  • DNA, Viral
  • Endotoxins