title: Hepatitis B and C virus-induced hepatitis: Apoptosis, autophagy, and unfolded protein response.
abstract: OBJECTIVE
  To investigate the co-incidence of apoptosis, autophagy, and unfolded protein response (UPR) in hepatitis B (HBV) and C (HCV) infected hepatocytes.
  
  
  METHODS
  We performed immunofluorescence confocal microscopy on 10 liver biopsies from HBV and HCV patients and tissue microarrays of HBV positive liver samples. We used specific antibodies for LC3β, cleaved caspase-3, BIP (GRP78), and XBP1 to detect autophagy, apoptosis and UPR, respectively. Anti-HCV NS3 and anti-HBs antibodies were also used to confirm infection. We performed triple blind counting of events to determine the co-incidence of autophagy (LC3β punctuate), apoptosis (cleaved caspase-3), and unfolded protein response (GRP78) with HBV and HCV infection in hepatocytes. All statistical analyses were performed using SPSS software for Windows (Version 16 SPSS Inc, Chicago, IL, United States). P-values < 0.05 were considered statistically significant. Statistical analyses were performed with Mann-Whitney test to compare incidence rates for autophagy, apoptosis, and UPR in HBV- and HCV-infected cells and adjacent non-infected cells.
  
  
  RESULTS
  Our results showed that infection of hepatocytes with either HBV and HCV induces significant increase (P < 0.001) in apoptosis (cleavage of caspase-3), autophagy (LC3β punctate), and UPR (increase in GRP78 expression) in the HCV- and HBV-infected cells, as compared to non-infected cells of the same biopsy sections. Our tissue microarray immunohistochemical expression analysis of LC3β in HBV(Neg) and HBV(Pos) revealed that majority of HBV-infected hepatocytes display strong positive staining for LC3β. Interestingly, although XBP splicing in HBV-infected cells was significantly higher (P < 0.05), our analyses show a slight increase of XBP splicing was in HCV-infected cells (P > 0.05). Furthermore, our evaluation of patients with HBV and HCV infection based on stage and grade of the liver diseases revealed no correlation between these pathological findings and induction of apoptosis, autophagy, and UPR.
  
  
  CONCLUSIONS
  The results of this study indicate that HCV and HBV infection activates apoptosis, autophagy and UPR, but slightly differently by each virus. Further studies are warranted to elucidate the interconnections between these pathways in relation to pathology of HCV and HBV in the liver tissue.
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Behzad
authors.initials: B
authors.lastname: Yeganeh
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Adel
authors.initials: A
authors.lastname: Rezaei Moghadam
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Javad
authors.initials: J
authors.lastname: Alizadeh
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Emilia
authors.initials: E
authors.lastname: Wiechec
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Seyed Moayed
authors.initials: SM
authors.lastname: Alavian
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Hashemi
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Bita
authors.initials: B
authors.lastname: Geramizadeh
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Afshin
authors.initials: A
authors.lastname: Samali
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Kamran
authors.initials: K
authors.lastname: Bagheri Lankarani
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Martin
authors.initials: M
authors.lastname: Post
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Payam
authors.initials: P
authors.lastname: Peymani
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Coombs
authors.affiliation: Behzad Yeganeh, Martin Post, Department of Physiology and Experimental Medicine, Hospital for Sick Children Research Institute, University of Toronto, Toronto M5G 0A4, Canada.
authors.forename: Saeid
authors.initials: S
authors.lastname: Ghavami
mesh_terms: D017209:Apoptosis; D001343:Autophagy; D015415:Biomarkers / Q000032:analysis; D001706:Biopsy; D053148:Caspase 3 / Q000032:analysis; D000091342:Endoplasmic Reticulum Chaperone BiP; D005455:Fluorescent Antibody Technique; D006509:Hepatitis B / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D006526:Hepatitis C / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D022781:Hepatocytes / Q000737:chemistry / Q000473:pathology* / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000473:pathology* / Q000821:virology; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000032:analysis; D012189:Retrospective Studies; D012720:Severity of Illness Index; D046888:Tissue Array Analysis; D056811:Unfolded Protein Response; D000071717:X-Box Binding Protein 1 / Q000032:analysis
chemical_list: D015415:Biomarkers; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; C502070:MAP1LC3B protein, human; D008869:Microtubule-Associated Proteins; D000071717:X-Box Binding Protein 1; C000605543:XBP1 protein, human; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: Cell death; Cell fate; Endoplasmic reticulum stress; Hepatocyte; Viral infection