title: Inhibitory effect of HMGN2 protein on human hepatitis B virus expression and replication in the HepG2.2.15 cell line.
abstract: Natural killer (NK) cells and cytolytic T lymphocytes (CTL) have been implicated as important effectors of antiviral defense. We previously isolated a novel antibacterial polypeptide, which was identified as high mobility group nucleosomal-binding domain 2 (HMGN2), from human mononuclear leukocytes. This study examined the antiviral activity of HMGN2 against human hepatitis virus B. HMGN2 was isolated and purified from the acid soluble proteins of the human THP-1 cell line, and identified by mass spectrum, Western blot and antibacterial assay. The hepatitis B virus (HBV)-transfected HepG2.2.15 cell line was used in the in vitro assay system. In the range of 1-100 microg/ml HMGN2, no cytotoxicity for HepG2.2.15 cells was detected by MTT assay. When incubated with HMGN2 at 1-100 microg/ml for 72 or 144 h, there was a significant reduction in hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) expression, which were detected by ELISA, and a significant reduction in HBV DNA copies, which was determined by the real time quantitative PCR, in the supernatant of HepG2.2.15 cells. Northern and Southern blot analysis also showed that the levels of the HBV 3.5 kb and the 2.4/2.1 kb mRNA species and HBV replicative intermediate DNA were significantly reduced in the HMGN2-treated HepG2.2.15 cells. These results indicated that HMGN2 protein could markedly inhibit HBV protein expression and replication in vitro.
authors.affiliation: State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Feng
authors.forename: Fang
authors.initials: F
authors.lastname: He
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Wu
authors.forename: Ning
authors.initials: N
authors.lastname: Huang
authors.forename: Hong
authors.initials: H
authors.lastname: Tang
authors.forename: Xiangli
authors.initials: X
authors.lastname: Kong
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Junju
authors.initials: J
authors.lastname: Lu
authors.forename: Qianming
authors.initials: Q
authors.lastname: Chen
authors.forename: Boyao
authors.initials: B
authors.lastname: Wang
mesh_terms: D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D002470:Cell Survival; D004279:DNA, Viral / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D024242:HMGN2 Protein / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis; D006513:Hepatitis B e Antigens / Q000096:biosynthesis; D006515:Hepatitis B virus / Q000187:drug effects* / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D004279:DNA, Viral; D024242:HMGN2 Protein; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012333:RNA, Messenger; D012367:RNA, Viral