Cyclosporin A inhibits Hepatitis B and Hepatitis D Virus entry by Cyclophilin-independent interference with the NTCP receptor
Shirin Nkongoloa, 1,
Yi Ni a, 1,
Florian A. Lempp a,
Christina Kaufman a, b,
Thomas Lindner b,
Katharina Esser-Nobis a,
Volker Lohmann a,
Walter Mier b,
Stefan Mehrle a,
Stephan Urban a, Corresponding author contact information, E-mail the corresponding author, E-mail the corresponding author
a Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, D 69120 Heidelberg, Germany
b Department of Nuclear Medicine, University Hospital Heidelberg, D 69120, Heidelberg, Germany
Abstract
Background & aims
Chronic hepatitis B and hepatitis D are global health problems caused by the human hepatitis B and hepatitis D virus. The myristoylated preS1 domain of the large envelope protein mediates specific binding to hepatocytes by sodium taurocholate co-transporting polypeptide (NTCP). NTCP is a bile salt transporter known to be inhibited by Cyclosporin A. This study aimed to characterize the effect of Cyclosporin A on HBV/HDV infection.
Methods
HepaRG cells, primary human hepatocytes and susceptible NTCP-expressing hepatoma cell lines were applied for infection experiments. The mode of action of Cyclosporin A was studied by comparing the effect of different inhibitors, cyclophilin A/B/C-silenced cell lines as well as NTCP variants and mutants. Bile salt transporter and HBV receptor functions were investigated by taurocholate uptake and quantification of HBVpreS binding.
Results
Cyclosporin A inhibited hepatitis B and D virus infections during and - less pronounced - prior to virus inoculation. Binding of HBVpreS to NTCP was blocked by Cyclosporin A concentrations at 8M. An NTCP variant deficient in HBVpreS binding but competent for bile salt transport showed resistance to Cyclosporin A. Silencing of cyclophilins A/B/C did not abrogate transporter and receptor inhibition. In contrast, tacrolimus, a cyclophilin-independent calcineurin inhibitor, was inactive.
Conclusions
HBV and HDV entry via sodium taurocholate co-transporting polypeptide is inhibited by Cyclosporin A. The interaction between the drug and the viral receptor is direct and overlaps with a functional binding site of the preS1 domain, which mediates viral entry.
Abbreviations
HBV, hepatitis B virus;
HBsAg, hepatitis B surface antigen;
HDV, hepatitis delta virus;
NTCP, sodium taurocholate co-transporting polypeptide;
TC, taurocholate;
CsA, Cyclosporin A;
HCV, hepatitis C virus;
hNTCP, human sodium taurocholate co-transporting polypeptide;
mNTCP, mouse sodium taurocholate co-transporting polypeptide;
GFP, green fluorescent protein;
PHH, primary human hepatocytes;
DMSO, dimethyl sulfoxide;
MyrB, Myrcludex B;
HBeAg, hepatitis B virus early antigen;
IF, immunofluorescence;
PBS, phosphate-buffered saline;
HBcAg, hepatitis B core antigen;
DAPI, 4′,6-diamidin-2-phenylindol;
MFI, mean geometric fluorescence intensity 作者: StephenW 时间: 2013-12-9 14:26