A rapid transfer of virions coated with heparan sulfate from the ECM to CD151 defines an early step in the human papillomavirus infection cascade.

Human Papillomaviruses (HPVs) are the underlying cause of several types of cancer; albeit, they are mostly known for their association with cervical carcinoma. The virions reach their target cells through a break in the epithelial barrier. After binding to heparan sulfate (HS) of the extracellular matrix (ECM), they are recruited via actin-dependent mechanisms to the cell surface, where they co-internalize with the entry factor CD151. The in vivo occurring active recruitment from the ECM to the cell surface may be bypassed in cell culture, where virions reach the cell surface simply by passive diffusion. To specifically investigate these early events of the infection cascade, we use HaCaT keratinocytes as they produce a robust ECM enabling abundant virion binding to ECM components such as HS before transfer to cell surface receptors and infection. Employing microscopy, we focus on the basal membrane that for virions is difficult to access by diffusion. We block the active recruitment from ECM attachment sites to the cell body, release the blocking, and monitor the association of virions with CD151 or HS. We observe quick virion recruitment from the ECM to the cell body within 15 min. During recruitment, virions associate with the tetraspanin CD151 present at the cell border or at filopodia. These virions are decorated with HS, which they lose in the next few hours, presumably prior to endocytosis. Our observations reveal a rapid step in the HPV infection cascade: the transfer of HS-coated virions from the ECM to CD151. This step is too fast to account for the asynchronous uptake of HPVs, which is likely driven by glycan and capsid processing.