Hepatitis B virus (HBV) infection can cause chronic liver disease, which is associated with increased risk of liver cirrhosis, liver failure, and liver cancer. Clearance of HBV infection requires effective HBV-specific immunity; however, the immunological mechanisms that determine the development of effective HBV-specific immunity are poorly understood. Dendritic cells (DC) play a pivotal role in the regulation of antiviral immunity. Here, we investigated the interaction between HBV surface antigen (HBsAg), the main envelope glycoprotein of HBV, and BDCA1+ myeloid dendritic cells (mDC). Exposure of peripheral blood-derived BDCA1+ mDC to HBsAg resulted in strong DC maturation, cytokine production, and enhanced capacity to activate antigen-specific cytotoxic T cells (CTLs). By using neutralizing antibodies, crucial roles for CD14 and Toll-like receptor 4 (TLR4) in HBsAg-mediated BDCA1+ mDC maturation were identified. Concordantly, HBsAg-mediated DC maturation required fetal calf serum (FCS) or human plasma, naturally containing soluble CD14 (sCD14). Intriguingly, HBsAg-induced DC maturation was significantly reduced in umbilical cord blood plasma, which contained less sCD14 than adult plasma, indicating that sCD14 is an important host factor for recognition of HBsAg by DC and subsequent DC activation. A direct interaction between sCD14 and HBsAg was demonstrated by using enzyme-linked immunosorbent assay (ELISA). Moreover, sCD14-HBsAg complexes were detected both in vitro and in sera of HBV-infected patients. The abundance of sCD14-HBsAg complexes varied between chronic HBV disease stages and correlated with activation of BDCA1+ mDC in vivo. We conclude that HBsAg activates BDCA1+ DC via an sCD14-dependent mechanism. These findings provide important novel insights into the initiation of HBV-specific immunity and facilitate development of effective immunotherapeutic interventions for HBV.
IMPORTANCE Hepatitis B virus (HBV) infection is a significant health problem, as it causes progressive liver injury and liver cancer in patients with chronic HBV infection, which affects approximately 250 million individuals worldwide. Some of the infected adults and the majority of neonates fail to mount an effective immune response and consequently develop chronic infection. The viral and host factors involved in the initiation of effective HBV-specific immune responses remain poorly understood. Here we identified CD14 and TLR4 as receptors for HBsAg, the main HBV envelope antigen. HBsAg induced strong maturation of dendritic cells (DC), which have a central role in regulation of virus-specific immunity. These results provide essential novel insights into the mechanisms underlying the initiation of HBV-specific immunity. Intriguingly, since neonates have naturally low sCD14, the finding that serum-derived sCD14 is a crucial host factor for recognition of HBsAg by DC may have implications for immunity of neonates to HBV infection.
FOOTNOTES
Received 16 November 2015.
Accepted 8 April 2016.
Accepted manuscript posted online 20 April 2016.
Address correspondence to Nadine van Montfoort, nvanmontfoort{at}lumc.nl, or Andrea M. Woltman, a.woltman{at}erasmusmc.nl.
↵* Present address: Nadine van Montfoort, Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands; Harry L. A. Janssen, Toronto Centre for Liver Disease, Division of Gastroenterology, University Health Network, Toronto, ON, Canada.
Citation van Montfoort N, van der Aa E, van den Bosch A, Brouwers H, Vanwolleghem T, Janssen HLA, Javanbakht H, Buschow SI, Woltman AM. 2016. Hepatitis B virus surface antigen activates myeloid dendritic cells via a soluble CD14-dependent mechanism. J Virol 90:6187–6199. doi:10.1128/JVI.02903-15.
通过可溶性CD14依赖机制乙肝病毒表面抗原激活髓细胞树突状细胞
乙肝病毒表面抗原激活髓细胞树突状细胞通过可溶性CD14依赖机制乙肝病毒表面抗原激活髓细胞树突状细胞
纳丁面包车Montfoorta *，伊夫林·范德AAA，Aniek范登Boscha希尔德Brouwersa，托马斯Vanwolleghema，哈里L. A. Janssena *，哈桑Javanbakhtb，宋佳一Buschowa和Andrea M. Woltmana