Journal of Virology, July 2002, p. 6710-6717, Vol. 76, No. 13
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.13.6710-6717.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Polarized Release of Human Cytomegalovirus from Placental Trophoblasts
D. G. Hemmings* and L. J. Guilbert*
Department of Medical Microbiology and Immunology and the Perinatal Research Centre, University of Alberta, Edmonton, Alberta, Canada
Received 26 November 2001/
Accepted 21 March 2002
Human cytomegalovirus (HCMV) is a ubiquitous infectious pathogen that, when transmitted to the fetus in utero, can result in numerous sequelae, including late-onset sensorineural damage. The villous trophoblast, the cellular barrier between maternal blood and fetal tissue in the human placenta, is infected by HCMV in vivo. Primary trophoblasts cultured on impermeable surfaces can be infected by HCMV, but release of progeny virus is delayed and minimal. It is not known whether these epithelial cells when fully polarized can release HCMV and, if so, if release is from the basal membrane surface toward the fetus. We therefore ask whether, and in which direction, progeny virus release occurs from HCMV-infected trophoblasts cultured on semipermeable (3.0-µm-pore-size) membranes that allow functional polarization. We show that infectious HCMV readily diffuses across cell-free 3.0-µm-pore-size membranes and that apical infection of confluent and multilayered trophoblasts cultured on these membranes reaches cells at the membrane surface. Using two different infection and culture protocols, we found that up to 20% of progeny virus is released but that <1% of released virus is detected in the basal culture chamber. These results suggest that very little, if any, HCMV is released from an infected villous trophoblast into the villous stroma where the virus could ultimately infect the fetus.
* Corresponding author. Present address for D. G. Hemmings: Department of Obstetrics and Gynaecology and Perinatal Research Centre, University of Alberta, 232 HMRC, University of Alberta, Edmonton, Alberta, Canada T6G 2S2. Phone: (780) 492-8562. Fax: (780) 492-1308. E-mail: denise.hemmings{at}ualberta.ca. Mailing address for L. J. Guilbert: 625 HMRC, University of Alberta, Edmonton, Alberta, Canada T6G 2S2. Phone: (780) 492-8562. Fax: (780) 492-9829. E-mail: larry.guilbert{at}ualbert.ca.
Journal of Virology, July 2002, p. 6710-6717, Vol. 76, No. 13
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.13.6710-6717.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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Copyright © 2002 by the American Society for Microbiology. All rights reserved.