Despite the prevalence of HPV16 in invasive cervical cancers, an in vitro system capable of producing infectious HPV16 is lacking. The organotypic (raft) culture system has allowed for the study of the entire differentiation-dependent life cycle of human papillomaviruses (HPVs). However, the use of this system with the prototype HPV16-containing cell line, W12, has failed to yield infectious virus. Our laboratory has introduced clinically derived HPV16(114/B) genomic DNA into primary keratinocytes, where it subsequently recircularized and maintained episomally at 50-100 copies per cell. Virion morphogenesis occurred after epithelial stratification and differentiation in raft culture. HPV16 virions were isolated that were able to infect keratinocytes in vitro. Infection was neutralized by monoclonal antibodies raised against HPV16 but not by monoclonal antibodies known to neutralize other HPV types.
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