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回复 baobao7676 的帖子
您的解释是错误的.
“马赛克状”分布在一个感染的肝细胞内的:
乙肝病毒抗原: HBsAg, HBcAg
与乙肝病核酸: cccDNA, HBVDNA, HBV RNA
In situ analysis of intrahepatic virological events
in chronic hepatitis B virus infection
Xiaonan Zhang,1 Wei Lu,2 Ye Zheng,3 Weixia Wang,1 Lu Bai,1,4 Liang Chen,2 Yanling Feng,3 Zhanqing Zhang,2
and Zhenghong Yuan1,4
1Research Unit, Shanghai Public Health Clinical Center, Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai, China. 2Department of Hepatology and
3Department of Clinical Pathology, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China. 4Key Laboratory of Medical Molecular Virology at the School of Basic Medical Sciences,
Shanghai Medical College, Fudan University, Shanghai, China.
Persistent hepatitis B virus (HBV) infection is established by the formation of an intranuclear pool of covalently closed
circular DNA (cccDNA) in the liver. Very little is known about the intrahepatic distribution of HBV cccDNA in infected patients,
particularly at the single-cell level. Here, we established a highly sensitive and specific ISH assay for the detection of HBV
RNA, DNA, and cccDNA. The specificity of our cccDNA probe set was confirmed by its strict intranuclear signal and by a series
of Southern blot analyses. Use of our in situ assay in conjunction with IHC or immunofluorescence uncovered a surprisingly
mosaic distribution of viral antigens and nucleic acids. Most strikingly, a mutually exclusive pattern was found between HBV
surface antigen–positive (HBsA-positive) and HBV DNA– and cccDNA-positive cells. A longitudinal observation of patients
over a 1-year period of adeforvir therapy confirmed the persistence of a nuclear reservoir of viral DNA, although cytoplasmic
DNA was effectively depleted in these individuals. In conclusion, our method for detecting viral nucleic acids, including
cccDNA, with single-cell resolution provides a means for monitoring intrahepatic virological events in chronic HBV infection.
More important, our observations unravel the complexity of the HBV life cycle in vivo. |
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