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J. Virol., 06 1995, 3258-3264, Vol 69, No. 6
C Volpers, F Unckell, P Schirmacher, RE Streeck and M Sapp
Infection of cells by human papillomaviruses (HPVs) associated with
malignant genital lesions has not been studied because of the lack of an in
vitro system and the unavailability of virions. We have now used virus-like
particles (VLPs) of HPV type 33 to analyze the initial events in the
interaction of the HPV capsid with cell lines. Binding of VLPs to HeLa
cells was observed in biochemical assays and by immunofluorescence. VLP
binding was inhibited by antisera raised against VLPs but not by monoclonal
antibodies recognizing either L1 or L2 epitopes accessible on VLPs. Under
saturating conditions, approximately 2 x 10(4) VLPs were bound per cell,
with a dissociation constant of about 100 pM. VLPs composed of L1 alone
bound as well as VLPs composed of both capsid proteins, indicating that L2
is not required for initial binding. VLPs dissociated into capsomers did
not bind, demonstrating that intercapsomer contacts are required. Neither
capsomers nor simian virus 40 virions competed with VLP binding. Uptake of
VLPs by small and smooth endocytic vesicles was demonstrated by
immunoelectron microscopy. Cellular binding of VLPs was sensitive to
trypsin but not to sialidase, N-glycosidase, or octyl-beta-D-
glycopyranoside treatment, suggesting that a cell surface protein is
involved in the VLP binding. Cell lines originating from a variety of
tissues and organisms as distantly related as insects and humans bound VLPs
with similar efficiency and specificity. Therefore, the putative receptor
mediating VLP attachment should be highly conserved and cannot be
responsible for the species and tissue specificity of HPVs.
Copyright © 1995, American Society for Microbiology
Binding and internalization of human papillomavirus type 33 virus-like particles by eukaryotic cells
Institut fur Medizinische Mikrobiologie und Hygiene, Universitat Mainz, Germany.
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