title: NKP30-B7-H6 Interaction Aggravates Hepatocyte Damage through Up-Regulation of Interleukin-32 Expression in Hepatitis B Virus-Related Acute-On-Chronic Liver Failure.
abstract: OBJECTIVE
  Previous work conducted by our group has shown that the accumulation of hepatic natural killer (NK) cells and the up-regulation of natural cytotoxicity receptors (NKP30 and NKP46) on NK cells from patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) were correlated with disease progression in HBV-ACLF. The natural cytotoxicity receptors expressed on NK cells are believed to be probable candidates involved in the NK cell-mediated hepatocyte damage in HBV-ACLF. However, the underlying mechanisms remain to be elucidated. In the present study, we aimed to discover the role of NKP30-B7-H6 interaction in NK cells-mediated hepatocyte damage in HBV-ACLF.
  
  
  METHODS
  Hepatic expressions of B7-H6 and interleukin-32 (IL-32) were examined by immunochemistry staining in samples from patients with HBV-ACLF or mild chronic hepatitis B (CHB). The cytotoxicity of NK-92 cell against target cells (Huh-7 and LO2) was evaluated by CCK8 assay. Expression of IL-32 in liver NK cell, T cells and NK-92 cell line was detected by the flow cytometric analysis. The effect of IL-32 on the apoptosis of Huh7 cells was evaluated using Annexin V/PI staining analysis.
  
  
  RESULTS
  An enhancement of hepatic B7-H6 and IL-32 expression was associated with the severity of liver injury in HBV-ACLF. And there was a positive association between hepatic B7-H6 and IL-32 expression. Expressions of IL-32 in liver NK cells and T cells were increased in HBV-ACLF patients. In vitro NK-92 cells are highly capable of killing the high B7-H6 expressing Huh7 cells and B7-H6-tansfected hepatocyte line LO2 cells dependent on NKP30 and B7-H6 interaction. Furthermore, NK-92 cells exhibited elevated IL-32 expression when stimulated with anti-NKP30 antibodies or when co-cultured with Huh7 cells. IL-32 can induce the apoptosis of Huh7 cells in a dose-dependent manner.
  
  
  CONCLUSIONS
  Our results suggest that NKP30-B7-H6 interaction can aggravate hepatocyte damage, probably through up-regulation of IL-32 expression in HBV-ACLF.
authors.affiliation: Department of Blood Transfusion, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Preterm Birth Prevention and Treatment Research Unit, Department of Obstetrics, Guangzhou Women and Children's Medical Center, Guangzhou, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Bao
authors.affiliation: Department of infectious disease, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: Xingfei
authors.initials: X
authors.lastname: Pan
authors.affiliation: Department of Blood Transfusion, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Blood Transfusion, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Sihong
authors.initials: S
authors.lastname: Liao
authors.affiliation: Department of oncology, Zhangqiu People's Hospital, Jinan, China.
authors.forename: Xicheng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Guoying
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Blood Transfusion, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China; Department of Hematology, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
authors.forename: Dongjun
authors.initials: D
authors.lastname: Lin
mesh_terms: D065290:Acute-On-Chronic Liver Failure / Q000378:metabolism* / Q000821:virology; D000328:Adult; D017209:Apoptosis / Q000187:drug effects; D060887:B7 Antigens / Q000378:metabolism*; D045744:Cell Line, Tumor; D005260:Female; D006509:Hepatitis B / Q000150:complications / Q000378:metabolism* / Q000821:virology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism* / Q000821:virology; D006801:Humans; D007378:Interleukins / Q000378:metabolism* / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D055628:Natural Cytotoxicity Triggering Receptor 3 / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology*; D055815:Young Adult
chemical_list: D060887:B7 Antigens; C496971:IL32 protein, human; D007378:Interleukins; C431710:NCR3 protein, human; C542363:NCR3LG1 protein, human; D055628:Natural Cytotoxicity Triggering Receptor 3