Hepatitis B virus replication is upregulated in proliferated peripheral blood lymphocytes

Abstract

Increasing evidence indicates that the hepatitis B virus (HBV) replicates in peripheral blood mononuclear cells (PBMCs), but at a low level. The present study aimed to establish a reliable and sensitive method that effectively detects HBV viral products for monitoring antiviral therapy, organ transplantation screening, and diagnosing occult HBV infection. In the present study, PBMCs (obtained from six healthy volunteers) were inoculated with HBV, and cultured with phytohemagglutinin (PHA) and interleukin‑2 (IL‑2) to stimulate cell proliferation. PBMCs were harvested, and quantitative detection of HBV DNA in cell suspension and intracellular hepatitis B surface antigen (HBsAg) was conducted on days 0, 1, 6 and 12, respectively. In situ hybridization, immunohistochemistry and reverse transcription‑polymerase chain reaction (RT‑PCR) were performed to analyze the HBV infection. The results demonstrated that HBV DNA increased concurrently with proliferation of PBMCs isolated from three of six healthy volunteers, and the mean number of PBMCs on day 12 was 13.61 times higher than the initially seeded cell number (P<0.01). The mean copies of HBV DNA at day 12 were 2.98 times higher compared with initial levels (P<0.05). Furthermore, intracellular HBsAg levels increased concurrently with proliferation of PBMCs in one group of cultured PBMCs, which was accompanied by increased HBV DNA levels. In addition, HBV nucleic acids were detected in PBMCs using in situ hybridization. Intracellular HBsAg was observed in PBMCs and HBV RNA was also detected by RT‑PCR. The present study demonstrated that HBV replicates in proliferating PBMCs, which were induced by PHA and IL‑2. This method offers a novel investigative tool to detect HBV infection in PBMCs and to monitor the course of HBV infection.

Comparison of Demographic, Epidemiological, Immunological, and Clinical Characteristics of Patients with HIV Mono-infection Versus Patients Co-infected with HCV or/and HBV: A Serbian Cohort Study

“…In the last 20 years numerous studies on the impact of HBV co-infection in the natural course of HIV infection have been carried out [57][58][59]. The finding that HBV although primarily hepatotropic also displays tropism for lymphoid cells, increased the likelihood of interaction of HIV and HBV at the cellular level, in patients with HIV/HBV coinfection [60,61]. In addition to the CD4+ T cell subpopulation, HBV also replicates in PBMC subsets of HBV/HIV-1 patients [62,63].…”

Impact of Hepatitis B Virus Genetic Variation, Integration, and Lymphotropism in Antiviral Treatment and Oncogenesis

“…In fact, Lee et al, Chemin et al, and Trippler et al have independently demonstrated that the virus is universally present in multiple immune cell subsets derived from HBV-infected individuals [ 122 , 123 , 124 ]. Studies by Yan et al and Huang et al reported that naïve primary PBMCs and hematopoietic stem cells, respectively, can be infected in vitro with clinically derived HBV [ 125 , 126 ]. Our recent work demonstrated that HBV derived from the lymphoid reservoir (PBMCs) can be stimulated with mitogens that induce an increase in multiple viral replication markers such as HBV DNA, cccDNA, and RNA both intracellularly and extracellularly [ 127 ].…”

Molecular diagnosis of citrin deficiency in an infant with intrahepatic cholestasis: identification of a 21.7kb gross deletion that completely silences the transcriptional and translational expression of the affected SLC25A13 allele

“…PBLs were isolated from 1ml of fresh EDTA-anticoagulant peripheral venous blood with lymphocyte separation medium (LSM, Ficoll-hypaque, density at 20°C: 1.077-1.080g/ml; MP Biomedicals, USA) and washed twice with 1× phosphate buffer saline (PBS). The PBLs were then adjusted to a density of 3-5 ×10 5 cells/ml by diluting with the above complete medium in the presence of 5μg/ml of PHA (Sigma, USA) and 10 ng/ml of recombinant IL-2 (Peprotech, USA) [ 54 , 62 , 63 ]. For subcultures, the high-density cultures of PBLs were diluted with the above stimulative medium down to an appropriate seeding density every 2-3 days.…”