Inhibition of the binding of HCV serum particles to human hepatocytes by E1E2-specific D32.10 monoclonal antibody.
The aim of this study was to determine the inhibition of binding activity of the monoclonal antibody (mAb) D32.10 which recognizes a highly conserved discontinuous antigenic determinant (E1:297-306, E2:480-494, and E2:613-621) expressed on the surface of serum-derived HCV particles (HCVsp) of genotypes 1a, 1b, 2a, and 3a. To this end, an in vitro direct cell-binding assay based on the attachment of radiolabeled HCVsp was developed, and Scatchard plots were used to analyze ligand-receptor binding data. HCV adsorption was also assessed by quantitating cell-associated viral RNA by a real-time RT-PCR method. Saturable concentration-dependent specific binding of HCVsp to Huh-7 or HepaRG cells was demonstrated. The Scatchard transformed data showed two-site interaction for Huh-7 and proliferative HepaRG cells: the high-affinity binding sites (K(d1) = 0.1-0.5 microg/ml) and the low-affinity binding sites (K(d1) = 5-10 microg/ml), and one-site high-affinity binding model between E1E2/D32.10-positive HCVsp and hepatocyte-like differentiated HepaRG cells. The E1E2-specific mAb D32.10 inhibited efficiently (>60%) and selectively the binding with an IC(50) <or=0.5 microg/ml in all the experimental approaches using serum HCV of genotype either 3 or 1b. This supports the involvement of the E1E2/D32.10 discontinuous antigenic determinant in the interactions between human hepatocytes and HCVsp, and suggests that D32.10-like antibodies present in sera from patients infected with HCV could play a protective role.
Ndongo
Ndiémé
N
Institut National de la Santé et de la Recherche Médicale, U871, Lyon, France.
Rechoum
Yassine
Y
De Sequeira
Sylvie
S
Zoulim
Fabien
F
Trépo
Christian
C
Drouet
Emmanuel
E
Petit
Marie-Anne
MA
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D016174:Hepacivirus / Q000276:immunology* / Q000502:physiology*; D018937:Hepatitis C Antibodies / Q000276:immunology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D020128:Inhibitory Concentration 50; D014759:Viral Envelope Proteins / Q000276:immunology; D053585:Virus Attachment
D000911:Antibodies, Monoclonal; C087035:E1 protein, Hepatitis C virus; D018937:Hepatitis C Antibodies; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus