Virus-neutralizing monoclonal antibody to a conserved epitope on the duck hepatitis B virus pre-S protein.
In this study we used duck hepatitis B virus (DHBV)-infected Pekin ducks and heron hepatitis B virus (HHBV)-infected heron tissue to search for epitopes responsible for virus neutralization on pre-S proteins. Monoclonal antibodies were produced by immunizing mice with purified DHBV particles. Of 10 anti-DHBV specific hybridomas obtained, 1 was selected for this study. This monoclonal antibody recognized in both DHBV-infected livers and viremic sera a major (36-kilodalton) protein and several minor pre-S proteins in all seven virus strains used. In contrast, pre-S proteins of HHBV-infected tissue or viremic sera did not react. Thus, the monoclonal antibody recognizes a highly conserved DHBV pre-S epitope. For mapping of the epitope, polypeptides from different regions of the DHBV pre-S/S gene were expressed in Escherichia coli and used as the substrate for immunoblotting. The epitope was delimited to a sequence of approximately 23 amino acids within the pre-S region, which is highly conserved in four cloned DHBV isolates and coincides with the main antigenic domain as predicted by computer algorithms. In in vitro neutralization assays performed with primary duck hepatocyte cultures, the antibody reduced DHBV infectivity by approximately 75%. These data demonstrate a conserved epitope of the DHBV pre-S protein which is located on the surface of the viral envelope and is recognized by virus-neutralizing antibodies.
Laboratoire de Recherche sur le hepatites, Institut National de la Santé et de la Recherche Médicale U 271, Lyon, France.
V
V
Lambert
D
D
Fernholz
R
R
Sprengel
I
I
Fourel
G
G
Deléage
G
G
Wildner
C
C
Peyret
C
C
Trépo
L
L
Cova
H
H
Will
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000276:immunology*; D004279:DNA, Viral / Q000032:analysis; D004372:Ducks; D004770:Enterovirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D015611:Hepatitis B Virus, Duck / Q000276:immunology* / Q000302:isolation & purification; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting; D008099:Liver / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011498:Protein Precursors / Q000276:immunology*; D012984:Software
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004279:DNA, Viral; D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen