No abstract

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 bloodderived 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. Hepatitis B virus (HBV) is a double-stranded DNA (dsDNA) virus that is transmitted via blood and specifically infects hepatocytes. It can cause chronic liver disease and progressive liver injury leading to increased risk of liver cirrhosis, liver failure, and liver cancer (1). The current estimated prevalence of HBV infection is 248 million individuals globally (2). Although the initiation of an effective antiviral immune response is paramount for resolving HBV infection (3), the early steps in the recognition of the virus by immune cells and the functional consequences of this interaction remain to be resolved.A pivotal role for dendritic cells (DC) is anticipated, because these cells play a central role in the orchestration of antiviral immunity due to the expression of a wide variety of different pathogen recognition receptors (PRR) and their unique capacity to initiate virus-specific cytotoxic T cell (CTL) responses (4, 5). Among the different DC subsets, BDCA1 ϩ myeloid DC (mDC) are of particular interest for HBV-specific immunity, since hepatitis B surface antigen (HBsAg)-positive mDC were detected in liver (6) an...

Chronic hepatitis B virus (HBV) infection results from inadequate HBV-specific immunity. BDCA3+ dendritic cells (DCs) are professional antigen presenting cells considered to be important for antiviral responses because of specific characteristics, including high interferon-λ production. BDCA3+ DCs may thus also have a role in the immune response against HBV, and immunotherapeutic strategies aiming to activate DCs, including BDCA3+ DCs, in patient livers may represent an interesting treatment option for chronic HBV. However, neither the effect of chronic hepatitis B (CHB) infection on the frequency and function of BDCA3+ DCs in liver and blood, nor the effect of the viral surface protein (HBsAg) that is abundantly present in blood of infected individuals are known. Here, we provide an overview of BDCA3+ DC frequency and functional capacity in CHB patients. We find that intrahepatic BDCA3+ DC numbers are increased in CHB patients. BDCA3+ DCs from patient blood are not more mature at steady state, but display an impaired capacity to mature and to produce interferon-λ upon polyI:C stimulation. Furthermore, in vitro experiments exposing blood and intrahepatic BDCA3+ DCs to the viral envelope protein HBsAg demonstrate that HBsAg does not directly induce phenotypical maturation of BDCA3+ DCs, but may reduce IFN-λ production via an indirect unknown mechanism. These results suggest that BDCA3+ DCs are available in the blood and on site in HBV infected livers, but measures may need to be taken to revive their function for DC-targeted therapy.

Keywords: BDCA3 r CD34 + -derived dendritic cells r GM-CSF/IL-4/TNF-β-driven culture system r Human r IFNAdditional supporting information may be found in the online version of this article at the publisher's web-site

Myeloid dendritic cells, including BDCA3hi DCs and BDCA1 + DCs (hereafter dubbed DC1 and DC2 for clarity), play a pivotal role in the induction and regulation of immune responses. Interestingly, a fraction of DC2 also express low to intermediate levels of BDCA3. It is unknown whether BDCA3 + DC2 also share other traits with DC1 that are absent in BDCA3 À DC2 and/or whether BDCA3 expression renders DC2 functionally distinct from their BDCA3-lacking counterparts. Here, we used expression analysis on a predefined set of immunology-related genes to determine divergence between BDCA3-positive and BDCA3-negative DC2 and their relation to bona fide BDCA3 hi DC1. Results showed that mRNA fingerprints of BDCA3 + DC2 and BDCA3 À DC2 are very similar, and clearly distinct from that of DC1.Differences in mRNA expression, however, were observed between BDCA3 + DC2 and BDCA3 À DC2 that pointed toward a more activated status of BDCA3 + DC2. In line with this, higher steady state maturation marker expression and TLR-induced maturation marker expression and inflammatory cytokine production by BDCA3 + DC2 were observed. This dataset provides insight into the relationship between myeloid DC populations and contributes to further understanding of DC immunobiology.