title: Effect of amniotic fluid on cationic lipid mediated transfection and retroviral infection.
abstract: In preparation for foetal gene therapy by intra-amniotic gene application, we have investigated the effect of amniotic fluid on several gene transfer systems. In vitro lipofection of embryonically derived 3T3 cells by several of the tested cationic lipids decreases in the presence of human amniotic fluid, while two formulations, Lipid 67 and Tfx-50, remain highly active. As some body fluids are known to inactivate most retroviral vectors, we investigated the influence of amniotic fluid on the efficiency of infection of 3T3 cells by murine leukaemia virus (MoMLV)-based vectors, including amphotropic and ecotropic retrovirus, and a vesicular stomatitis virus G (VSV-G) glycoprotein pseudotyped retroviral vector. All showed a decrease of infectivity from 21 to 56% in the presence of amniotic fluid. The ecotropic retrovirus is the most infectious under normal conditions as well as in amniotic fluid. Our results suggest that intra-amniotic injection may allow efficient gene transfer using either amniotic fluid-resistant cationic lipids or ecotropic retroviral vectors in a murine in vivo model for foetal gene therapy. The VSV-G-pseudotyped vector, although displaying a decrease of infectivity, remains of great interest for gene delivery, because of its broad host range and because of the high virus titers achievable. Finally, a baculovirus-based vector shows no decrease of its infectivity and does not seem to be affected by amniotic fluid but has only low infectivity on the tested foetal fibroblast cell line.
authors.affiliation: Department of Biochemistry and Molecular Genetics, Imperial College, Medical School at St Mary's London, UK.
authors.forename: A M
authors.initials: AM
authors.lastname: Douar
authors.forename: M
authors.initials: M
authors.lastname: Themis
authors.forename: V
authors.initials: V
authors.lastname: Sandig
authors.forename: T
authors.initials: T
authors.lastname: Friedmann
authors.forename: C
authors.initials: C
authors.lastname: Coutelle
mesh_terms: D016475:3T3 Cells; D000653:Amniotic Fluid; D000818:Animals; D002412:Cations; D002460:Cell Line; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D008055:Lipids; D008562:Membrane Glycoproteins; D051379:Mice; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011247:Pregnancy; D012190:Retroviridae; D014162:Transfection / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics
chemical_list: D002412:Cations; C028391:G protein, vesicular stomatitis virus; D008055:Lipids; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins