Antiviral activity of an intracellularly expressed single-chain antibody fragment directed against the murine leukemia virus capsid protein.
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.
Marin
M
M
Institut de Génétique Moléculaire de Montpellier, UMR 5535 CNRS/IFR 24, France.
Pelegrin-Zurilla
M
M
Bachrach
E
E
Noël
D
D
Brockly
F
F
Piechaczyk
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000914:Antibodies, Viral / Q000378:metabolism*; D001483:Base Sequence; D002213:Capsid / Q000276:immunology*; D002457:Cell Extracts; D002460:Cell Line; D003001:Cloning, Molecular; D015151:Immunoblotting; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D009052:Leukemia Virus, Murine / Q000254:growth & development / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism*; D051381:Rats; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D002457:Cell Extracts; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments; D015682:Retroviridae Proteins, Oncogenic; D014759:Viral Envelope Proteins; C060711:gp30 envelope transmembrane protein, Bovine leukemia virus