Internalization of human rhinovirus 14 into HeLa and ICAM-1-transfected BHK cells.
Virus adsorption and uptake of human rhinovirus 14 (HRV14) were studied with HeLa cells and baby hamster kidney (BHK) cells which were transfected with the HRV14 receptor intercellular adhesion molecule-1 (ICAM-1). Transmission electron microscopy of HeLa cells revealed that HRV14 was internalized via clathrin-coated pits and -coated vesicles. A minority of virus particles also used uncoated vesicles for entry. The internalization showed the characteristics of receptor-mediated endocytosis. Presence of the carboxylic ionophore monensin inhibited viral uncoating, indicating a pH-dependent entry mechanism. The expression of ICAM-1 on the surface of the ICAM-1 transfected baby hamster kidney cells (BHK-ICAM cells) allowed extensive virus adsorption and internalization through membrane channels. Virus particles were lined up in these channels like pearls on a string, but did not induce a productive infection. Although ICAM-1 was expressed to the same degree on BHK-ICAM and HeLa cells, HRV14 induced neither viral protein and RNA syntheses nor infectious virus progeny in BHK-ICAM cells. ICAM-1 on the transfected BHK cells was a functional active receptor as it rendered these cells permissive to coxsackievirus A21. These results suggest that HRV14 uptake into BHK-ICAM cells is blocked directly in or shortly after its final step of internalization, the uncoating. Our findings underline that the receptor ICAM-1 determines virus uptake into cells, however, is not sufficient to confer susceptibility of BHK cells to HRV14 infection.
Grunert
H P
HP
Institut für Klinische und Experimentelle Virologie, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Germany.
Wolf
K U
KU
Langner
K D
KD
Sawitzky
D
D
Habermehl
K O
KO
Zeichhardt
H
H
D000818:Animals; D006224:Cricetinae; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000502:physiology*; D007668:Kidney / Q000821:virology; D012229:Rhinovirus / Q000502:physiology*; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis
D014764:Viral Proteins; D018799:Intercellular Adhesion Molecule-1