摘要
目的探索影响树鼩体内长期感染HBV的因素。方法77只新生期和49只非新生期树鼩，分别以不同的感染源和注射方式接种HBV，部分动物加以辅助措施。接种后定期获取动物血和肝组织，应用荧光定量聚合酶链反应、酶联免疫吸附试验、时间分辨免疫荧光分析、Southemblot和dotblot、免疫组织化学染色、电子和光学显微镜等多种方法，长期动态观察动物体内HBV感染标志的消长。结果新生期接种的树鼩中6只确定、7只疑似为持续感染，非新生期接种的树鼩无一为持续感染。单份的HBV患者或树嗣血清为效果较好的感染源；皮下注射为较好的接种途径；接种用血清先经纯化有一定的辅助作用，联合检测树嗣血清的HBV免疫学标志和肝组织的HBVDNA为较好的检测方案。结论不同的接种年龄、感染源、接种方法、辅助措施可影响树鼩体内慢性感染HBV的效果，合适的检测方案有助于判断和预测树鼩的感染状态。
Objective To determine the methods for establishing an in vivo model of long-term hepatitis B virus （HBV） infection in the Chinese tree shrew （Tupaia belangeri chinensis）. Methods Seventyseven neonate （1-3 days old） and 49 young adult （2 weeks to 1 year old） tree shrews were inoculated with different HBV sources （chronic hepatitis B （CHB） human patient serum, single or pooled; HBV- infected tree shrew serum, single only; HBV-infeeted HepG2.2.15 cells＇ culture medium supernatant; HBV genome-transfected HepG2.2.15 cells＇ culture medium supernatant） through various routes of injection （subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen）. Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy. Results Among the neonatal group of tree shrews, six （7.8%） were confirmed as HBV-infected for more than 72 （up to 228） weeks after inoculation and another seven （9.1%） were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most eases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA. Conclusion In order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2012年第9期654-658,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（30660171） 广西自然科学基金（0447087）
关键词
肝炎病毒
乙型
模型
动物
树鼩
Hepatitis B virus
Models, animal
Tree shrews
