Persistent hepatitis B virus (HBV) infection is established by the formation of an intranuclear pool of covalently closed circular DNA (cccDNA) in the liver. So far so little is known about the intrahepatic distribution of HBV cccDNA in infected patients, particularly at the single-cell level.
A joint research group leaded by Prof. Zhenghong Yuan with many cohorts at Fudan established a highly sensitive and specific ISH assay for the detection of HBV RNA, DNA, and cccDNA. The specificity of their cccDNA probe set was confirmed by its strict intranuclear signal and by a series of Southern blot analyses. By Using 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 antigenâpositive (HBsA-positive) and HBV DNAâ 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. conclusion could be made is this method for detecting viral nucleic acids, including cccDNA, with single-cell resolution provides a vehicle for monitoring intrahepatic virological events in chronic HBV infection. More importantly this observations unravel the complexity of the HBV life cycle in vivo. On 2 Feb.2016 the study was made open on "Journal of Clinical Investigation" on-line version, thanks to a joint collaborativ efforts by Key Lab of Virology of Shanghai Medical College, Fudan and Research Divison of Shanghai Public Clinical Center, Fudan Univeristy, This in-depth scientific investigation has drawn a wide attention among academic cohorts, recommanded as a highlight report with keynote comment by Prof.Stephen Locarnini and Peter Revill, it will soon be offically published on JCI March Volumn.
