title: HPV binding assay to Laminin-332/integrin α6β4 on human keratinocytes.
abstract: Human papillomaviruses (HPVs) have been shown to bind to Laminin-332 (Ln-332) on the extracellular matrix (ECM) secreted by human keratinocytes. The assay described here is an important tool to study HPV receptor binding to the ECM. The assay can also be modified to study the receptors required for HPV infection and for binding to tissues. We previously showed that Ln-332 is essential for the binding of HPV11 to human keratinocytes and that infectious entry of HPV11 requires α6β4 integrin for the transfer of HPV11 from ECM to host cells (Culp et al., J Virol 80:8940-8950, 2006). We also demonstrated that several of the high-risk HPV types (16, 18, 31 and 45) bind to Ln-332 and/or other components of the ECM in vitro (Broutian et al., J Gen Virol 91:531-540, 2010). The exact binding and internalization mechanism(s) for HPV are still under investigation. A better understanding of these mechanisms will aid in the design of therapeutics against HPVs and ultimately help prevent many cancers. In this chapter, we describe the HPV binding assay to Ln-332/integrin α6β4 on human keratinocytes (ECM). We also present data and suggestions for modifying the assay for testing the specificity of HPV for receptors (by blocking receptors) and binding to human tissues (basement membrane, BM) in order to study binding mechanisms.
authors.affiliation: Department of Pathology, The Jake Gittlen Laboratories for Cancer Research, Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Brendle
authors.forename: Neil D
authors.initials: ND
authors.lastname: Christensen
mesh_terms: D001681:Biological Assay / Q000379:methods*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002460:Cell Line; D002584:Cervix Uteri / Q000378:metabolism / Q000821:virology; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D039161:Integrin alpha6beta4 / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism*; D027383:Papillomaviridae / Q000378:metabolism*; D011485:Protein Binding; D011991:Receptors, Virus / Q000378:metabolism; D014771:Virion / Q000378:metabolism; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D039161:Integrin alpha6beta4; D011991:Receptors, Virus; D000097794:Kalinin