Feng Yun, He Fang, Zhang Ping, Wu Qi, Huang Ning, Tang Hong, Kong Xiangli, Li Yan, Lu Junju, Chen Qianming, Wang Boyao
State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China.
Antiviral Res. 2009 Mar;81(3):277-82. doi: 10.1016/j.antiviral.2008.12.011. Epub 2009 Jan 14.
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.
自然杀伤（NK）细胞和细胞毒性T淋巴细胞（CTL）被认为是抗病毒防御的重要效应细胞。我们之前从人单核白细胞中分离出一种新型抗菌多肽，经鉴定为高迁移率族核小体结合域2（HMGN2）。本研究检测了HMGN2对人乙型肝炎病毒的抗病毒活性。从人THP-1细胞系的酸溶性蛋白中分离并纯化HMGN2，并通过质谱、蛋白质印迹和抗菌试验进行鉴定。体外试验系统采用乙型肝炎病毒（HBV）转染的HepG2.2.15细胞系。在1 - 100μg/ml的HMGN2浓度范围内，MTT试验未检测到对HepG2.2.15细胞的细胞毒性。当HepG2.2.15细胞与1 - 100μg/ml的HMGN2孵育72或144小时后，ELISA检测显示乙型肝炎e抗原（HBeAg）和乙型肝炎表面抗原（HBsAg）表达显著降低，实时定量PCR测定显示HepG2.2.15细胞上清液中的HBV DNA拷贝数显著减少。Northern和Southern印迹分析还表明，在HMGN2处理的HepG2.2.15细胞中，HBV 3.5 kb和2.4/2.1 kb mRNA种类以及HBV复制中间体DNA的水平显著降低。这些结果表明，HMGN2蛋白在体外可显著抑制HBV蛋白表达和复制。
