PreS1-specific binding proteins as potential receptors for hepatitis B virus in human hepatocytes.
Cellular receptors play an important role in viral pathogenesis. Until now, there has been no reliable information on the receptor(s) for hepatitis B virus (HBV). Therefore, we attempted to identify specific receptors in human hepatocytes using an immunological approach. Anti-idiotypic (Ab2) antibodies were raised in rabbits against our monoclonal antibody (MAb1) F35.25. MAb1 F35.25 (i) recognized the hepatocyte receptor binding site on HBV (located between amino acid residues 21 and 47 of the preS1 sequence) and (ii) blocked the attachment of preS1-positive HBV particles to human hepatocytes. The presence of Ab2 antibodies in rabbit sera was determined by the ability of antisera to inhibit Id (Ab1)/antigen (HBV) recognition. Affinity-purified Ab2 IgGs to F35.25 represented an internal image for the preS1 domain 12-53. Our present studies indicate that Ab2 IgGs to F35.25 (i) recognized the membrane-associated structure of the preS1-specific HBV receptor in a HepG2 cell binding assay, as visualized by immunoenzymatic staining; (ii) strongly bound to a major 35-kDa component and to three other related proteins of 50, 43, and 40 kDa in extracts of HepG2 cells; and (iii) reacted with several soluble and membrane-associated proteins in normal human liver cells. The binding was insensitive to reduction. All preS1 binding proteins were V8 protease sensitive and endoglycosidase H resistant. The 35-kDa species was trypsin resistant and generated a band of 32 kDa by endoglycosidase F treatment. Together, our results suggest that the identified preS1-specific binding proteins may be involved in the putative complex structure of the hepatocyte receptor for HBV.
Petit
M A
MA
INSERM U 131, Clamart, France.
Capel
F
F
Dubanchet
S
S
Mabit
H
H
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D001665:Binding Sites / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology / Q000378:metabolism*; D006515:Hepatitis B virus / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000737:chemistry* / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism*; D011863:Radioimmunoassay; D011946:Receptors, Antigen / Q000276:immunology / Q000378:metabolism*; D011991:Receptors, Virus / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011946:Receptors, Antigen; D011991:Receptors, Virus; C030500:hepatitis B surface antigen receptor; C050806:presurface protein 1, hepatitis B surface antigen