Immunoadhesins containing pre-S domains of hepatitis B virus large envelope protein are secreted and inhibit virus infection.
Hepatitis B virus (HBV) replication produces three envelope proteins (L, M, and S) that have a common C terminus. L, the largest, contains a domain, pre-S1, not present on M. Similarly M contains a domain, pre-S2, not present on S. The pre-S1 region has important functions in the HBV life cycle. Thus, as an approach to studying these roles, the pre-S1 and/or pre-S2 sequences of HBV (serotype adw2, genotype A) were expressed as N-terminal fusions to the Fc domain of a rabbit immunoglobulin G chain. Such proteins, known as immunoadhesins (IA), were highly expressed following transfection of cultured cells and, when the pre-S1 region was present, >80% were secreted. The IA were myristoylated at a glycine penultimate to the N terminus, although mutation studies showed that this modification was not needed for secretion. As few as 30 amino acids from the N terminus of pre-S1 were both necessary and sufficient to drive secretion of IA. Even expression of pre-S1 plus pre-S2, in the absence of an immunoglobulin chain, led to efficient secretion. Overall, these studies demonstrate an unexpected ability of the N terminus of pre-S1 to promote protein secretion. In addition, some of these secreted IA, at nanomolar concentrations, inhibited infection of primary human hepatocytes either by hepatitis delta virus (HDV), a subviral agent that uses HBV envelope proteins, or HBV. These IA have potential to be part of antiviral therapies against chronic HDV and HBV, and may help understand the attachment and entry mechanisms used by these important human pathogens.
Chai
Ning
N
Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111-2497, USA.
Gudima
Severin
S
Chang
Jinhong
J
Taylor
John
J
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D003698:Hepatitis Delta Virus / Q000254:growth & development*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D007155:Immunologic Factors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology
D000998:Antiviral Agents; D006514:Hepatitis B Surface Antigens; D007141:Immunoglobulin Fc Fragments; D007155:Immunologic Factors; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen; C050807:presurface protein 2, hepatitis B surface antigen