Carboxy-fluorescein diacetate, succinimidyl ester labeled papillomavirus virus-like particles fluoresce after internalization and interact with heparan sulfate for binding and entry
2003 (English)In: Virology, ISSN 0042-6822, E-ISSN 1096-0341, Vol. 310, no 1, 163-172 p.Article in journal (Refereed) Published
Human papillomaviruses (HPVs) infect epithelial cells and are associated with genital carcinoma. Most epithelial cell lines express cell-surface glycosaminoglycans (GAGs) usually found attached to the protein core of proteoglycans. Our aim was to study how GAGs influenced HPV entry. Using a human keratinocyte cell line (HaCaT), preincubation of HPV virus-like particles (VLPs) with GAGs showed a dose-dependent inhibition of binding. The IC(50) (50% inhibition) was only 0.5 microg/ml for heparin, 1 microg/ml for dextran sulfate, and 5-10 microg/ml for heparan sulfate from mucosal origin. Mutated chinese hamster ovary (CHO) cell lines lacking heparan sulfate or all GAGs were unable to bind HPV VLPs. Here we also report a method to study internalization by using VLPs labeled with carboxy-fluorescein diacetate, succinimidyl ester, a fluorochrome that is only activated after cell entry. Pretreatment of labeled HPV VLPs with heparin inhibited uptake, suggesting a primary interaction between HPV and cell-surface heparan sulfate.
Place, publisher, year, edition, pages
2003. Vol. 310, no 1, 163-172 p.
Papillomavirus; CFDA SE; Heparan sulfate; Uptake, Receptor
IdentifiersURN: urn:nbn:se:umu:diva-2394DOI: 10.1016/S0042-6822(03)00114-4PubMedID: 12788640OAI: oai:DiVA.org:umu-2394DiVA: diva2:140371