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Journal of Virology, July 2000, p. 6050-6057, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Retargeting the Coxsackievirus and Adenovirus Receptor to the
Apical Surface of Polarized Epithelial Cells Reveals the Glycocalyx as
a Barrier to Adenovirus-Mediated Gene Transfer
Raymond J.
Pickles,1,*
Jill A.
Fahrner,1
JenniElizabeth M.
Petrella,2
Richard C.
Boucher,1 and
Jeffrey
M.
Bergelson2
CF/Pulmonary Research and Treatment Center,
University of North Carolina at Chapel Hill, Chapel Hill, North
Carolina 27599-7248,1 and Division of
Immunologic and Infectious Diseases, Children's Hospital of
Philadelphia, Philadelphia, Pennsylvania 191042
Received 2 February 2000/Accepted 25 March 2000
Lumenal delivery of adenovirus vectors (AdV) results in inefficient
gene transfer to human airway epithelium. The human coxsackievirus and
adenovirus receptor (hCAR) was detected by immunofluorescence selectively at the basolateral surfaces of freshly excised human airway
epithelial cells, suggesting that the absence of apical hCAR
constitutes a barrier to adenovirus-mediated gene delivery in vivo. In
transfected polarized Madin-Darby canine kidney cells, wild-type hCAR
was expressed selectively at the basolateral membrane, whereas hCAR
lacking the transmembrane and/or cytoplasmic domains was expressed on
both the basolateral and apical membranes. Cells expressing apical hCAR
still were not efficiently transduced by AdV applied to the apical
surface. However, after the cells were treated with agents that remove
components of the apical surface glycocalyx, AdV transduction occurred.
These results indicate that adenovirus can infect via receptors located
at the apical cell membrane but that the glycocalyx impedes interaction
of AdV with apical receptors.
*
Corresponding author. Mailing address: CF/Pulmonary
Research and Treatment Center, UNC School of Medicine, 7129 Thurston-Bowles, University of North Carolina at Chapel Hill,
Chapel Hill, NC 27599-7248. Phone: (919) 966-7044. Fax: (919)
966-7524. E-mail: branston{at}med.unc.edu.
Journal of Virology, July 2000, p. 6050-6057, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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