title: Characterization of human papillomavirus type 16 pseudovirus containing histones.
abstract: Pseudoviruses (PsVs) that encapsidate a reporter plasmid DNA have been used as surrogates for native human papillomavirus (HPV), whose continuous production is technically difficult. HPV PsVs have been designed to form capsids made up of the major capsid protein L1 and the minor capsid proteins L2. HPV PsVs have been produced in 293TT cells transfected with plasmid expressing L1 and L2 protein and plasmid containing the reporter gene. Several studies have suggested that naturally occurring HPV virions contain cellular histones, and histones have also been identified in mature HPV PsVs. However, the effect of the histones on the properties of the PsVs has not been investigated. Using heparin chromatography, we separated mature HPV type 16 PsVs into three fractions (I, II, and III) according to their heparin-binding affinities.
  
  
  
  The amounts of cellular histone and cellular nucleotides per PsV were found to increase in the order fraction I, II and III. It appeared that PsVs in fraction I contains just small amount of cellular histone in Western blot analysis. The proportions of the three fractions in PsV preparations were 83.4, 7.5, and 9.1 % for fraction I, II, and III PsVs, respectively. In the electron microscope PsVs in fraction I appeared to have a more condensed structure than those in fractions II and III. Under the electron microscope fraction II and III PsVs appeared to be covered by substantial amounts of cellular histone while there was no visible histone covering PsVs of fraction I. Also the levels of reporter gene expression in infections of fraction II and III PsVs to 293TT cells were significantly lower than those in infections of fraction I PsV, and fraction II and III particles had significantly reduced immunogenicity.
  
  
  
  Our findings suggest that the involvement of large amounts of cellular histones during PsV formation interferes with the structural integrity of the PsVs and affects their immunogenicity. The fraction I particle therefore has the most suitable characteristics for use as an HPV PsV.
authors: Laboratory of Virology, College of Pharmacy, Chung-Ang University, 84 Heukseok-Ro, Dongjak-Gu, Seoul, 06974, South Korea.
authors: Hyoung Jin
authors: HJ
authors: Kim
authors: Laboratory of Virology, College of Pharmacy, Chung-Ang University, 84 Heukseok-Ro, Dongjak-Gu, Seoul, 06974, South Korea.
authors: Hye-Lim
authors: HL
authors: Kwag
authors: Laboratory of Virology, College of Pharmacy, Chung-Ang University, 84 Heukseok-Ro, Dongjak-Gu, Seoul, 06974, South Korea. hongjink@cau.ac.kr.
authors: Hong-Jin
authors: HJ
authors: Kim
mesh_terms: D017930:Genes, Reporter / Q000235:genetics*; D024861:Genetic Enhancement / Q000379:methods; D006657:Histones / Q000378:metabolism*; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D013045:Species Specificity; D058615:Synthetic Biology; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D006657:Histones; D014764:Viral Proteins
keywords: Heparin chromatography; Histone; Human papillomavirus; Pseudovirion