Lentiviral hepatitis B pseudotype entry requires sodium taurocholate co-transporting polypeptide and additional hepatocyte-specific factors

Abstract

Hepatitis B virus (HBV) is one of the world's major unconquered infections, resulting in progressive liver disease, and current treatments rarely cure infection. A limitation to discovering new therapies is our limited knowledge of HBV entry and dissemination pathways that hinders the development of in vitro culture systems. To address this gap in our understanding we optimized the genesis of infectious lentiviral pseudoparticles (HBVpps). The recent discovery that the bile salt transporter sodium taurocholate co-transporting polypeptide (NTCP) acts as a receptor for HBV enabled us to assess the receptor dependency of HBVpp infection. HBVpps preferentially infect hepatoma cells expressing NTCP, whereas other non-liver cells engineered to express NTCP do not support infection, suggesting that additional hepatocyte-specific factors are required for HBVpp internalization. These results highlight the value of the HBVpp system to dissect the pathways of HBV entry and dissemination. Hepatitis B virus (HBV) infection is a major global health problem frequently resulting in progressive, degenerative liver disease, including cirrhosis and hepatocellular carcinoma. Despite the availability of a safe and effective vaccine, and a range of nucleoside analogue antivirals, ,240 million people are chronically infected with HBV (El-Serag, 2012). The high rate of viral turnover has resulted in vaccinerelated escape mutants and drug resistance (Billioud et al., 2012;Locarnini & Yuen, 2010). Therefore, development of safe, efficient antiviral strategies remains a key challenge for the treatment of HBV.

Solute Carrier NTCP Regulates Innate Antiviral Immune Responses Targeting Hepatitis C Virus Infection of Hepatocytes

“…ISGs also restrict HCV cell-to-cell transmission (Meredith et al., 2014), consistent with our finding that NTCP contributes to HCV cell-to-cell spread (Figures 1E–1G). It should be noted that NTCP expression did not appear to affect HCV entry in a recent Huh7 cell line model (Meredith et al., 2016). Huh7.5.1 cells are derived from Huh7.5 cells, which differ from Huh7 cells by a single point mutation in the retinoic acid-inducible gene-I (Bartenschlager and Pietschmann, 2005, Zhong et al., 2005).…”

Synchronized infection identifies early rate-limiting steps in the hepatitis B virus life cycle

“…HepG2-NTCP K7 cells and Huh-7 NTCP cells 21,60 were cultured in Dulbecco’s Modified Eagles Medium F12 (DMEM-F12) supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin. HepaRG cells were cultured in Williams E medium supplemented with 10% FBS, 50 U penicillin/streptomycin mL -1 , 5 μg human insulin mL -1 and 5×10 -7 M hydrocortisone hemisuccinate (Sigma).…”

Defined host factors support HBV infection in non‐hepatic 293T cells

“…Only human cell lines of hepatic origin reconstituted with hNTCP become HBV susceptible, and HBV replication was still restricted in NTCP‐ overexpressing non‐hepatic human HeLa cells and mouse hepatoma cell lines . In addition, human lung epithelial cells (A549) overexpressing NTCP also do not support HBV infection . These results suggested that NTCP expression alone is not sufficient to confer HBV infection and that additional liver‐specific factors are required.…”

“…35,36 In addition, human lung epithelial cells (A549) overexpressing NTCP also do not support HBV infection. 37 These results suggested that NTCP expression alone is not sufficient to confer HBV infection and that additional liver-specific factors are required.…”