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Journal of Virology, September 2001, p. 8772-8780, Vol. 75, No. 18
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.18.8772-8780.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Heparan Sulfate Glycosaminoglycans Are Receptors Sufficient
To Mediate the Initial Binding of Adenovirus Types 2 and
5
M. C.
Dechecchi,1
P.
Melotti,1
A.
Bonizzato,1
M.
Santacatterina,2
M.
Chilosi,2 and
G.
Cabrini1,*
Laboratory of Molecular Pathology, Cystic
Fibrosis Center,1 and Department of
Pathology, University of Verona,2 Verona,
Italy
Received 15 February 2001/Accepted 9 April 2001
Cell infection by adenovirus serotypes 2 and 5 (Ad2/5) initiates
with the attachment of Ad fiber to the coxsackievirus and Ad receptor
(CAR) followed by
v integrin-mediated entry. We recently demonstrated that heparan sulfate glycosaminoglycans (HS GAGs) expressed on cell surfaces are involved in the binding and infection of
Ad2/5 (M. C. Dechecchi, A. Tamanini, A. Bonizzato, and G. Cabrini, Virology 268:382-390, 2000). The role of HS GAGs was investigated using extracellular soluble domain 1 of CAR (sCAR-D1) and heparin as
soluble receptor analogues of CAR and HS GAGs in A549 and recombinant CHO cell lines with differential levels of expression of the two receptors and cultured to various densities. Complete inhibition of
binding and infection was obtained by preincubating Ad2/5 with both
heparin (10 µg/ml) and sCAR-D1 (200 µg/ml) in A549 cells. Partial
inhibition was observed when heparin and sCAR-D1 were preincubated
separately with Ad. The level of heparin-sensitive [3H]Ad2/5 binding doubled in sparse A549 cells (50 to
70,000 cells/cm2) with respect to that of cells grown to
confluence (200 to 300,000 cells/cm2), in parallel with
increased expression of HS GAGs. [3H]Ad2 bound to sparse
CAR-negative CHO cells expressing HS GAGs (CHO K1). No
[3H]Ad2 binding was observed in CHO K1 cells upon
competitive inhibition with heparin and in HS GAG-defective CHO A745,
D677, and E606 clones. HS-sensitive Ad2 infection was obtained in
CAR-negative sparse CHO K1 cells but not in CHO A745 cells, which
were permissive to infection only upon transfection with CAR.
These results demonstrate that HS GAGs are sufficient to mediate the
initial binding of Ad2/5.
*
Corresponding author. Mailing address: Laboratorio
Patologia Molecolare, Centro Regionale Fibrosi Cistica, Piazzale
Stefani, 1, 37126 Verona, Italy. Phone: 39-045-807 2364. Fax:
39-045-807 2840. E-mail: cabrini{at}linus.univr.it.
Journal of Virology, September 2001, p. 8772-8780, Vol. 75, No. 18
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.18.8772-8780.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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