The Impact of Hepatitis B co-infection on the HIV Reservoir

Hepatitis B virus (HBV) co-infection increases overall and liver-related mortality in people living with HIV, yet the relationship between these two viruses is still poorly understood. We determined whether levels of HIV were associated with markers of HBV infection, immune activation and inflammation or liver damage. We also designed an assay to sequence full-length HIV RNA in order to further validate our findings. HIV-HBV co-infected and HIV mono-infected participants, naïve to ART, were recruited in Bangkok, Thailand as part of a prospective observational cohort study. HIV proviruses were sequenced from peripheral blood (PB) CD4+ T cells using full-length individual proviral sequencing. Spearmans rank correlations tests were performed to determine associations. RNA extraction and cDNA synthesis protocols based on previously established Single Copy Assay and Single Genome Sequencing methods were optimised to yield a full-length HIV fragment. The 1337 and 285 HIV proviruses were sequenced from 19 HIV-HBV co-infected and 6 HIV mono-infected individuals respectively, although 2 co-infected and 1 mono-infected individual was excluded due to infection with non-subtype AE HIV virus. More genetically intact proviruses (22-1000 copies/106 cells, p=0.02) was observed in the co-infected individuals compared to the mono-infected. For the HIV-HBV cohort, the levels of soluble CD14, LPS and AST in the blood were significantly correlated with the frequency and the genetic diversity of intact HIV proviruses (p0.05). We were also able to present a preliminary HIV RNA sequencing assay to sequence ~90% of the HIV RNA genome with a single amplicon. During HIV-HBV co-infection, the frequency and genetic diversity of the intact proviruses within PB CD4+ T cells from the HIV-HBV co-infected individuals appears to be independent of levels of markers of HBV or liver damage, but may be associated with markers of immune activation.