Detection of Hepatitis B Virus (HBV) Genomes and HBV Drug Resistant Variants by Deep Sequencing Analysis of HBV Genomes in Immune Cell Subsets of HBV Mono-Infected and/or Human Immunodeficiency Virus Type-1 (HIV-1) and HBV Co-Infected Individuals

Abstract

The hepatitis B virus (HBV) and the human immunodeficiency virus type 1 (HIV-1) can infect cells of the lymphatic system. It is unknown whether HIV-1 co-infection impacts infection of peripheral blood mononuclear cell (PBMC) subsets by the HBV. Aims To compare the detection of HBV genomes and HBV sequences in unsorted PBMCs and subsets (i.e., CD4+ T, CD8+ T, CD14+ monocytes, CD19+ B, CD56+ NK cells) in HBV mono-infected vs. HBV/HIV-1 co-infected individuals. Methods Total PBMC and subsets isolated from 14 HBV mono-infected (4/14 before and after anti-HBV therapy) and 6 HBV/HIV-1 co-infected individuals (5/6 consistently on dual active anti-HBV/HIV therapy) were tested for HBV genomes, including replication indicative HBV covalently closed circular (ccc)-DNA, by nested PCR/nucleic hybridization and/or quantitative PCR. In CD4+, and/or CD56+ subsets from two HBV monoinfected cases, the HBV polymerase/overlapping surface region was analyzed by next generation sequencing. Results All analyzed whole PBMC from HBV monoinfected and HBV/HIV coinfected individuals were HBV genome positive. Similarly, HBV DNA was detected in all target PBMC subsets regardless of antiviral therapy, but was absent from the CD4+ T cell subset from all HBV/HIV-1 positive cases (P<0.04). In the CD4+ and CD56+ subset of 2 HBV monoinfected cases on tenofovir therapy, mutations at residues associated with drug resistance and/or immune escape (i.e., G145R) were detected in a minor percentage of the population. Summary HBV genomes and drug resistant variants were detectable in PBMC subsets from HBV mono-infected individuals. The HBV replicates in PBMC subsets of HBV/HIV-1 patients except the CD4+ T cell subpopulation.

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

“…Indeed, the prior study by Shi et al demonstrated the possible infectivity of hematopoietic stem cells with HBV [ 138 ]. This could also explain the distribution of HBV amongst the different immune cell subtypes, including natural killer cells, monocytes, B-lymphocytes, and T-lymphocytes as previously reported [ 122 , 123 , 124 ].…”

“…Important HBV replicative intermediates and infection markers have been identified in extrahepatic lymphoid tissues including the spleen, lymphoid nodes, and lymphoid cells [ 54 , 56 , 115 , 116 , 117 , 118 , 119 , 120 , 121 ]. 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 ].…”

“…The woodchuck and woodchuck hepatitis virus (WHV), a useful animal model system for hepadnaviruses, has established lymphoid infection by WHV [ 111 , 112 , 113 , 114 ]. Several research groups have suggested the detection of viral nucleic acids including HBV cccDNA, RNA, and DNA as well as viral integration within lymphoid cells [ 54 , 56 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 ]. Furthermore, direct infection of lymphoid cells with clinically derived HBV has been achieved in vitro [ 125 , 126 ].…”

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

“…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]. Some studies have shown that the HBV-X protein (HBX) accelerates existing HIV replication, inducing transcription of the HIV LTR sequences [64].…”

Prevalence and molecular characterization of occult hepatitis B virus in pregnant women from Gondar, Ethiopia

“…Moreover, as a result of the HIV epidemic in Sub-Saharan Africa and shared routes of transmission, HBV and HIV co-infection is common (5,(23)(24)(25). Although some HIV antiretroviral medications also have activity against HBV, patients on first-generation anti-HBV therapy with a lower genetic barrier to resistance (ie, lamivudine) may drive selection of HBV-resistant variants (5,(26)(27)(28). Our previous studies in an HIV-positive cohort from Gondar as well as a North American cohort of HIV-positive individuals (including 23 of African descent) with either CHB or OHB showed the frequent presence of S gene and DRMs (5,26,29).…”