CROI 2020 Abstract eBook

Abstract eBook

Poster Abstracts

611 A RHESUS MACAQUE MODEL OF CHRONIC HBV INFECTION FOR CURE RESEARCH Sreya Biswas 1 , Patrick Hashiguchi 1 , Jennifer Stanton 1 , Benjamin N. Bimber 1 , Ulrike Protzer 2 , Jonah Sacha 1 , Benjamin J. Burwitz 1 1 Oregon Health and Sciences University, Portland, OR, USA, 2 Technical University of Munich, Munich, Germany Background: Chronic HBV infection (CHB) is a major global health concern, affecting 247 million individuals worldwide and causing 887,000 deaths annually. CHB induces various degrees of liver damage and is strongly associated with the development of liver cirrhosis and hepatocellular carcinoma. Chimpanzees were the gold standard for primate HBV research, but are no longer available. Indeed, one of the major obstacles to the discovery of an HBV cure is the lack of a physiologically-relevant primate model. Based on our previous work showing that expression of the HBV receptor, human Na+- taurocholate co-transporting polypeptide (hNTCP), on rhesus macaque (RM) hepatocytes facilitates in vitro and in vivo HBV infection, we hypothesized that RM can be chronically infected with HBV. Methods: We treated three infant RM (<1-year-old) with an immunosuppression regimen consisting of daily tacrolimus and semi-monthly belatacept injections. Following initiation of this immunosuppression, we intravenously administered high-dose adenovirus expressing hNTCP. Seven days later we challenged all three RM intravenously with HBV (1 x 109 virions). Immunosuppression was tapered after 18 weeks of HBV infection. We have followed HBV infection in the blood and liver in these RM by qPCR, ELISA, and immunofluorescent microscopy over the course of 42 weeks. Results: We found persistently high levels of HBV plasma viremia (>1 x 105 copies/ml) accompanied by high levels of HBV surface (HBsAg) and envelope (HBeAg) antigens in blood for more than six months, the clinical definition of chronic HBV infection. In addition, high frequencies of HBV core antigen (HBcAg)- and HBsAg-positive hepatocytes were detected longitudinally in liver biopsies. Following immunosuppression tapering, two of the three animals maintained ongoing viral replication, indicating HBV immunotolerance. The set point HBV loads in these two animals correlated with the level of hNTCP expression in the liver by qPCR, indicating that hepatocyte target availability is the restricting factor in this model. Conclusion: Our data indicate that RM can be chronically infected with HBV and represent a promising model for the testing of emerging HBV curative therapies. Kathleen E. Stevens 1 , Eric C. Seaberg 2 , Katherine Cascino 1 , Mallory Witt 3 , Claudia Hawkins 4 , Bernard J. Macatangay 5 , Chloe Thio 1 1 Johns Hopkins University School of Medicine, Baltimore, MD, USA, 2 Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA, 3 Harbor–UCLA Medical Center, Torrance, CA, USA, 4 Northwestern University, Chicago, IL, USA, 5 University of Pittsburgh, Pittsburgh, PA, USA Background: Hepatitis B Virus (HBV) co-infection occurs in 5-20% of HIV- infected individuals globally. Prior studies found elevated proinflammatory cytokines are associated with spontaneous control (SC) of symptomatic HBV, but the majority of acute HBV infections are asymptomatic. A better understanding of cytokine profiles in early asymptomatic HBV will provide insight into immune responses important for natural resolution of infection, and may suggest immune targets for curative HBV therapies. Using men enrolled in the Multicenter AIDS Cohort Study (MACS) with asymptomatic incident HBV, we compared cytokine profiles between men who either had SC or developed chronic hepatitis B (CHB). Methods: Incident HBV occurred in 186 MACS men between 1985-2006 with available plasma samples. SC or CHB was determined by serological testing at 9-12 months after incident infection. Plasma samples at visits immediately before and after incident HBV (visits 1 and 2, respectively) were tested for 24 cytokines by electrochemiluminescence assay (Mesoscale Discovery, Rockville, MD), according to manufacturer's instructions. Non-parametric rank sum analyses were used to compare cytokine concentrations and intrasubject changes (visit 2/ visit 1). Results: Of 186 men, 18 were black (9.7%), 68 (37%) were HIV-infected (+)(8 on anti-HBV treatment at visit 1), and median age was 32 years (IQR=26-39). Twenty-four (12.9%, 14 HIV+) progressed to CHB, and 162 (87.1%, 54 HIV+) had SC. HIV infection was associated with greater odds of CHB (OR 2.8, P=0.02). At visit 1, HIV infection was associated with increased cytokine concentrations, including IL10, TNF, IP10, MIP1b, and IL18. 612 CYTOKINE PROFILES IN ASYMPTOMATIC ACUTE HEPATITIS B

Median time from incident HBV to visit 2 was 13.2 weeks. Overall, increased odds of CHB was significantly associated with elevated visit 2 levels of (OR per 10-fold increase in concentration): IL10 (OR 6.7, P<0.001), MIP1a (OR 25, P<0.001), IP10 (OR 8.3, P=0.001), MIP1b (OR 3.8, P=0.04), and IL18 (OR 7.7, P=0.04). In HIV-uninfected individuals, intrasubject increases (visit 2 fold- change over visit 1) in IL10, IP10, IL18, IL37, and MIP1a were significantly higher in those who developed CHB. In HIV-infected individuals, intrasubject increases in IL18 and TNF were higher in those who developed CHB (see Table). Conclusion: In contrast to previous reports, elevated cytokine profiles are not associated with SC in asymptomatic incident HBV, suggesting they are not major determinants of HBV SC.

Poster Abstracts

613 HBV-RELATED INFLAMMATION IS LINKED TO THE LEVEL OF GENETICALLY INTACT HIV PROVIRUSES Xiao Qian Wang 1 , Jennifer M. Zerbato 2 , Anchalee Avihingsanon 3 , Katie Fisher 1 , Bethany A. Horsburgh 1 , Timothy E. Schlub 4 , Ajantha Solomon 2 , Jennifer Audsley 2 , Kasha P. Singh 2 , Wei Zhao 2 , Megan Crane 5 , Sharon R. Lewin 2 , Sarah Palmer 1 1 The Westmead Institute for Medical Research, Westmead, NSW, Australia, 2 Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia, 3 HIV–NAT, Thai Red Cross AIDS Research Centre, Bangkok, Thailand, 4 The University of Sydney, Sydney, NSW, Australia, 5 Peter MacCallum Cancer Centre, Melbourne, VIC, Australia Background: Hepatitis B virus (HBV) coinfection increases overall and liver- related mortality in people living with HIV, even with the availability of HBV- active ART. HIV can persist in individuals in both defective and intact forms and both can contribute to persistent inflammation. We assessed the relationship between HIV proviral genomes and markers of inflammation in people living with HIV-HBV coinfection. Methods: HIV-HBV coinfected and HIV monoinfected participants, naïve to ART, were recruited in Bangkok, Thailand as part of a prospective observational cohort study. HIV subtype AE proviruses were sequenced from peripheral blood (PB) CD4+ T-cells using full-length individual proviral sequencing, covering 92% of the genome. Circulating markers of inflammation and microbial translocation were quantified by ELISA and bead arrays. Spearmans rank correlations tests were performed to determine associations. Results: 1008 and 222 HIV proviruses were sequenced from 18 HIV-HBV coinfected and 6 HIV monoinfected individuals respectively. The coinfected cohort had a significantly higher HIV viremia (p=0.03) and lower CD4+ T-cell count (p=0.007) than the monoinfected. A strong trend towards more intact proviruses (22-1000 copies/10 6 cells, p=0.055) was observed in the coinfected individuals. For the HIV-HBV cohort, the levels of soluble CD14 (sCD14), LPS and CXCL10 in the blood, markers of immune activation and/or inflammation, were significantly correlated with the frequency of intact HIV proviruses (p<0.01, p=0.04, p<0.01 respectively). sCD14 and CXCL10 were also correlated with the genetic diversity of the intact proviruses (p=0.03, for both). AST levels in blood, a marker of liver inflammation, and HIV DNA levels in the liver were also significantly correlated with the frequency of intact HIV proviruses in PB CD4+ T-cells (p=0.04, p=0.05 respectively). However, intact proviruses alone did not correlate with the number of PB CD4+ T-cells (p=0.2) but the inclusion of defective forms revealed a significant correlation with PB CD4+ T-cells (p=0.03). Conclusion: During HIV-HBV coinfection, the levels of PB CD4+ T-cells may be influenced by the amount of intact and defective proviruses they contain. However, the frequency and genetic diversity of the intact proviruses within blood-derived cells from the HIV-HBV coinfected individuals appears to be linked to inflammation and liver damage.

CROI 2020 221