Expression of vaccinia recombinant HPV 16 L1 and L2 ORF proteins in epithelial cells is sufficient for assembly of HPV virion-like particles

A recombinant vaccinia virus termed pLC201 VV was designed to coexpress the Li and L2 late genes of human papillomavirus type 16 (HPV16). Synthesis of the Li and L2 proteins occurred in cells infected with pLC201VV, and 40-nm virus-like particles with a density of 1.31 g/ml were produced in the nuclei of cells synthesizing both L1 and L2, but not in cells synthesizing either protein alone. Virus-like particles were partially purified from infected cells by sucrose gradient sedimentation and shown to consist of capsomeres similar to HPV and contain glycosylated L1 viral capsid protein. The production of HPV-like particles using recombinant vaccinia virus should be useful for biochemical studies and could provide a safe source of material for the development of a vaccine.