We have addressed the possibility that intracellularly expressed miniantibodies directed against the viral capsid protein can be used as antiretroviral agents in gene transfer experiments. R187 is a rat monoclonal antibody that has been reported to recognize the MuLV p30gag capsid polypeptide. We report here that it also binds to the Pr65gag precursor polyprotein. R187 has been cloned and expressed in the form of a single-chain variable fragment (scFv) that shows the same binding specificity as the parental antibody. When expressed intracellularly, the R187 scFv favors the production of viral particles showing reduced infectivity. It, however, exerts no detectable protective effect against infection. This was observed both when using replication-incompetent MuLV-derived vector and replication-competent wild-type MuLV. Although the intimate mechanism of the inhibition is not clear, this work raises the possibility that gene engineering of anti-capsid protein scFvs may offer an additional lead for gene therapy of severe retrovirus-linked diseases.
Antiviral activity of an intracellularly expressed single-chain antibody fragment directed against the murine leukemia virus capsid protein
Marin, M.; Pelegrin-Zurilla, M.; Bachrach, E.; Noel, D.; Brockly, F.; Piechaczyk, M.
Hum Gene Ther
2000-02-10 / vol 11 / pages 389-401
Animals; Amino Acid Sequence; Molecular Sequence Data; Base Sequence; Cell Line; Immunoblotting; Rats; Transfection; Cloning, Molecular; Antibodies, Monoclonal/metabolism; Antibodies, Viral/*metabolism; Capsid/*immunology; Cell Extracts; Immunoglobulin Variable Region/metabolism; Leukemia Virus, Murine/growth & development/*immunology; Peptide Fragments/*metabolism; Retroviridae Proteins, Oncogenic/*immunology; Viral Envelope Proteins/*immunology