title: Transduction of human trophoblastic cells by replication-deficient recombinant viral vectors. Promoting cellular differentiation affects virus entry.
abstract: We investigated the transfer of the lacZ reporter gene into human trophoblastic cells using herpes simplex virus and adeno-associated virus vectors. We used an established choriocarcinoma cell line (BeWo cells) that can be induced to terminally differentiate after treatment with cyclic-AMP. Our results demonstrate that transduction of trophoblastic cells by the herpes simplex virus vector, HSV.CMVlac, and the adeno-associated virus vector, AAV.CMVlac, is affected by cellular differentiation. Treatment of BeWo cells with cyclic-AMP reduced transduction by HSV.CMVlac but increased transduction by the AAV vector. In contrast, when BeWo cells were transfected with herpes simplex virus and adeno-associated virus plasmids, lacZ expression was not affected by treatment with cyclic-AMP. Southern blot analysis demonstrated 2.75 times less herpes simplex virus DNA in cyclic-AMP treated BeWo cells, but 2.0 to 7.4 times more adeno-associated virus DNA in treated cells. We conclude that inefficient transduction of differentiated trophoblastic cells with HSV.CMVlac is because of diminished viral entry, whereas cellular differentiation is associated with increased entry of AAV.CMVlac. These observations suggest that adeno-associated virus vectors may be used to modify trophoblast function and study placental physiology. Additionally, trophoblast differentiation leads to alterations in the mechanisms of virus uptake that may affect maternal-to-fetus transmission.
authors.affiliation: Department of Obstetrics and Gynecology, University of Pennsylvania Medical Center, Philadelphia, USA. sparry@obgyn.upenn.edu
authors.forename: S
authors.initials: S
authors.lastname: Parry
authors.forename: J
authors.initials: J
authors.lastname: Holder
authors.forename: M W
authors.initials: MW
authors.lastname: Halterman
authors.forename: M D
authors.initials: MD
authors.lastname: Weitzman
authors.forename: A R
authors.initials: AR
authors.lastname: Davis
authors.forename: H
authors.initials: H
authors.lastname: Federoff
authors.forename: J F
authors.initials: JF
authors.lastname: Strauss
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics; D015139:Blotting, Southern; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D000229:Dependovirus / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics; D006801:Humans; D014162:Transfection / Q000187:drug effects*; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects / Q000821:virology*; D014407:Tumor Cells, Cultured
chemical_list: D015124:8-Bromo Cyclic Adenosine Monophosphate