Hepatitis B Virus Dna in Peripheral–Blood Mononuclear Cells in Chronic Hepatitis B After Hbsag Clearance

Abstract

In this study, peripheral-blood mononuclear cells from patients with chronic hepatitis B and spontaneous or therapy-induced disappearance of HBsAg were examined for HBV DNA. Samples were evaluated by in situ hybridization and polymerase chain reaction both before and after clearance of HBsAg. By in situ hybridization, positive signals were observed in 2 of 13 samples collected after HBsAg loss, in 8 of 15 samples before HBsAg loss and in 0 of 4 control patients without serological markers of active or prior HBV infection. When polymerase chain reaction analyses were performed, HBV DNA was detected in 5 of 12 HBsAg-negative samples and 10 of 15 HBsAg-positive samples from the study group. Testing of mononuclear cells after disappearance of HBsAg revealed that two of eight patients were HBV DNA positive by in situ hybridization and by polymerase chain reaction, whereas two additional patients were positive by polymerase chain reaction alone. Mononuclear cell-associated HBV DNA was detected between 2 and 9 mo after the disappearance of circulating HBsAg by in situ hybridization and as long as 4 yr later by polymerase chain reaction. These data indicate that patients who have undergone HBsAg seroconversion may nonetheless harbor HBV DNA in their peripheral-blood mononuclear cells for prolonged periods.

Occult Lifelong Persistence of Infectious Hepadnavirus and Residual Liver Inflammation in Woodchucks Convalescent From Acute Viral Hepatitis

“…In general, our results are in agreement with these reports that demonstrate that lymphoid cells are the sites of active hepadnavirus replication and an important nonhepatocyte reservoir of the virus. [32][33][34][35][36] Because of many analogies between WHV and HBV and parallels in the profiles and outcomes of the induced diseases, our data should contribute to a better understanding of the natural history and biological implications of occult HBV infection in humans.…”

Molecular basis for persistent hepatitis B virus infection in the liver after clearance of serum hepatitis B surface antigen

“…Each patient was assessed by PCR for persistent HBV DNA in serum liver and peripheral blood mononuclear cells. One of 7 were HBV-DNApositive in the serum, 6 of 7 were HBV-DNA-positive in the liver, 8 and 4 of 7 were HBV-DNA-positive in the peripheral blood mononuclear cells 16 at the time of the repeat biopsy.…”

“…6, 1998 sterile water, and 200 to 500 ng of DNA was used for each PCR reaction. 16 RT-PCR. The RNA detection was performed both by a one-step method using Retrotherm RT TM (Epicentre Technology, Madison, WI), which has reverse transcriptase and thermostable DNA polymerase activity, and by a two-step method using Moloney murine leukemia virus reverse transcriptase (Gibco BRL, Gaithersburg, MD), followed by PCR amplification with Taq polymerase (Perkin Elmer Cetus, Norwalk, CT).…”

“…[8][9][10][11][12][13][14] In fact, HBV DNA can also be found in bodily secretions and peripheral blood mononuclear cells from patients with acute and chronic HBV infection after sustained loss of serum HBsAg. [15][16][17] Taken together, these studies suggest that following the loss of HBsAg from serum, HBV persists in a state of low-level replication or in a replication-competent state that can be reactivated to form infectious particles.The natural biology of the latent virus may be further investigated by assessing the transcriptional activity and the molecular form of the persistent HBV DNA. Using in situ hybridization, we were unable to detect hepatic HBV RNA in patients who had cleared serum HBsAg, and other investigators had little success using reverse-transcription (RT) PCR studies because of the inability to completely eradicate tissue HBV DNA.…”

“…[8][9][10][11][12][13][14] In fact, HBV DNA can also be found in bodily secretions and peripheral blood mononuclear cells from patients with acute and chronic HBV infection after sustained loss of serum HBsAg. [15][16][17] Taken together, these studies suggest that following the loss of HBsAg from serum, HBV persists in a state of low-level replication or in a replication-competent state that can be reactivated to form infectious particles.…”