Journal
JOURNAL OF VIRAL HEPATITIS
Volume 9, Issue 4, Pages 265-271Publisher
BLACKWELL PUBLISHING LTD
DOI: 10.1046/j.1365-2893.2002.00367.x
Keywords
covalently closed circular HBV-DNA; cell cultures; HBV-RNA; occult HBV infection
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Previously, we identified a defective hepatitis B virus (HBV) which contains a 183 nucleotide deletion in the PreS1 region of the viral genome affecting the S gene promoter in sera from hepatitis B surface antigen (HBsAg)-negative patients with serum HBV-DNA. The aim of this study was to analyse the infectivity of this mutant. Peripheral blood mononuclear cells (PBMC) from a healthy donor were incubated with serum samples from 2 HBsAg-negative patients with serum HBV-DNA (infected with wild-type and deletion mutant HBV), from an HBsAg carrier (infected with wild-type HBV) and from a healthy donor. After 1 week, HBV-DNA was detected by polymerase chain reaction (PCR) in all supernatants and cells incubated with the HBV-DNA-positive inocula. DNase and trypsin pretreatment confirmed intracellular localization of HBV-DNA in cells. HBV-RNA and covalently closed circular HBV-DNA were also detected in PBMC, indicating that the viral DNA infecting these cells was transcriptionally active. Deletion mutant and wild-type HBV were detected in the supernatants and cells infected with the two HBsAg-negative sera, while only wild-type HBV was detected in the supernatant and cells incubated with the serum from the HBsAg-carrier. In conclusion, this HBV deletion mutant can infect, replicate and release viral particles in in vitro infected PBMC.
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