Infected hematopoietic stem cells and with integrated HBV DNA generate defective T cells in chronic HBV infection patients

Abstract

A weak T-cell response plays a key role in the persistence of hepatitis B virus (HBV) infection. We aimed to confirm that T-cell defects in patients with chronic HBV infection are associated with HBV DNA infection of bone marrow (BM) hematopoietic stem cells (HSCs). Using reverse transcription polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), we observed the transcription of HBsAg coding genes and confirmed the integration of HBV DNA in CD34+ BM HSCs from chronic HBV infection patients. T cells were generated by coculturing the HSCs with delta-like ligand 1-expressing OP9 (OP9-DL1) cells. The phenotypes of the T cells were then evaluated by flow cytometric (FACS) analysis on days 14 and 25. The results demonstrated that fewer CD3+TCRaβ+ CD3+CD4+ and CD4+CD8+ T cells were generated from the HSCs of the patients than from the healthy controls, (P < 0.01) but the frequency of CD3+D8+ T cells was not significantly different between the two group (P > 0.05). In contrast, CD4+CD25+ T cells were more in the patient group than in healthy controls (P < 0.01) on both days 14 and 25. There were fewer CD3+CD4+/CD3+CD8+ cells in the patient group than in the healthy control group on day 25 (P < 0.05). Less proliferation and lower levels of IL-2 and IFN- γ were also observed in the patient group compared with the control group (P < 0.05).These data suggest that HBV DNA infected and integrated into the BM HSCs from patients with chronic HBV infection and that these BM HSCs generated defective T cells.

Detection of hepatitis b virus DNA in the blood of a stem cell donor after granulocyte colony‐stimulating factor treatment

“…(3) However, further studies are needed to link HBV reactivation and cell proliferation and to address a potential role of CD341 cells for the detectability of HBV DNA. (4) In accordance with previous observations, the presence of anti-HBs did not always provide protection from HBV reactivation. (5) At least in this case the presence of HBsAg mutations might have also contributed to the noneffectiveness of anti-HBs.…”

“…Because G‐CSF treatment induces proliferation of hepatocytes or hepatic progenitor cells in humans and other animals, it is tempting to speculate that here the G‐CSF treatment resulted in the proliferation of hepatocytes harboring HBV covalently closed circular DNA . However, further studies are needed to link HBV reactivation and cell proliferation and to address a potential role of CD34+ cells for the detectability of HBV DNA . In accordance with previous observations, the presence of anti‐HBs did not always provide protection from HBV reactivation .…”

Hepatitis B virus replication is upregulated in proliferated peripheral blood lymphocytes

“…Furthermore, it was reported that woodchuck HBV replication is upregulated in PBMCs with mitogen stimulation (19)(20)(21). Our previous studies also suggested that HBV gene expression is increased following expansion of bone marrow hematopoietic stem cells isolated from chronic hepatitis B patients, via a non-specific mitogen stimulus (22,23). Based on these findings, the aim of the present study was to establish a method to increase HBV replication in PBMCs via mitogen stimulation in vitro.…”

Persistent risk of HBV reactivation despite extensive lamivudine prophylaxis in haematopoietic stem cell transplant recipients who are anti-HBc-positive or HBV-negative recipients with an anti-HBc-positive donor

“…However, HBV-negative recipients receiving HSC from anti-HBc-positive donors had a 10.5% risk of developing HBV-related hepatitis after HSCT compared with recipients from HBV-negative donors in a country with a high level of HBV prevalence [46]. Stem cells from HBV-positive individuals were also shown to be a potential source of HBV [30]. In our study, 11 out of 15 allo-HSCT HBV-negative patients with an anti-HBcpositive/HBsAg-negative donor were treated with LMV.…”