title: HCV dsRNA-Activated Macrophages Inhibit HCV Replication in Hepatocytes.
abstract: BACKGROUND
  Macrophages play critical roles in innate immune response in the liver. Whether macrophages participate in liver innate immunity against HCV replication is poorly understood.
  
  
  OBJECTIVE
  The aim of this study was to investigate the role of macrophages in liver innate immunity against HCV replication.
  
  
  METHODS
  Freshly isolated monocytes were purified from peripheral blood of healthy adult donors. Macrophages refer to 7-day-cultured monocytes in vitro. A hepatoma cell line (Huh7) was infected with HCV JFH-1 to generate in vitro HCV infectious system. RT-PCR was used to determine HCV RNA and mRNA levels of genes expression. ELISA was used to measure the protein level of interferon-α (IFN-α) and western blot was used to determine protein expression level of Toll-like receptor 3 (TLR3).
  
  
  RESULTS
  HCV dsRNA induced the expression of type I IFN (IFN-α/β) in monocyte-derived macrophages. HCV dsRNA also induced the expression of TLR3 and IFN regulatory factor-7 (IRF-7), the key regulators of the IFN signaling pathway. When HCV JFH-1-infected Huh7 cells were co-cultured with macrophages activated with HCV dsRNA or incubated in media conditioned with supernatant (SN) from HCV dsRNA-activated macrophages, HCV replication was significantly suppressed. This macrophage SN action on HCV inhibition was mediated through type I IFN, which was evidenced by the observation that antibody to type I IFN receptor could neutralize the macrophages-mediated anti-HCV effect. The role of type I IFN in macrophages-mediated anti-HCV activity is further supported by the observation that HCV dsRNA-activated macrophages SN treatment induced the expression of several IFN-stimulated genes (ISGs), ISG15, ISG56, OAS-1, OAS-2, MxA and Viperin in HCV-infected Huh7 cells.
  
  
  CONCLUSIONS
  Macrophages may play an important role in liver innate immunity against HCV replication through a type I IFN-dependent mechanism.
authors: Department of Infectious Diseases, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China.
authors: Yizhong
authors: Y
authors: Wang
authors: Department of Pathology and Laboratory Medicine, School of Medicine, Temple University, Philadelphia, USA.
authors: Jieliang
authors: J
authors: Li
authors: Department of Pathology and Laboratory Medicine, School of Medicine, Temple University, Philadelphia, USA.
authors: Xu
authors: X
authors: Wang
authors: Department of Pathology and Laboratory Medicine, School of Medicine, Temple University, Philadelphia, USA.
authors: Yu
authors: Y
authors: Zhou
authors: Department of Infectious Diseases, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China.
authors: Ting
authors: T
authors: Zhang
authors: Department of Pathology and Laboratory Medicine, School of Medicine, Temple University, Philadelphia, USA.
authors: Wenzhe
authors: W
authors: Ho
keywords: Hepatitis C Virus; Interferon; Macrophages; Toll-Like Receptor 3