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Native Human Papillomavirus Production, Quantification, and Infectivity Analysis

  • Jennifer Biryukov
  • Linda Cruz
  • Eric J. Ryndock
  • Craig Meyers
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1249)

Abstract

In a natural infection, human papillomavirus (HPV) replicates in a stratified and differentiated epithelium. We have developed an in vitro organotypic raft culture system that allows researchers to study HPV in its natural environment. Not only does this system reproduce the differentiation-dependent replication cycle of HPV, but it also allows for the production of high titers of native HPV virions. Currently, much of the HPV research has been done utilizing synthetic particles produced in transfection systems. However, by production of native virions, this research can now be continued using native particles. This chapter presents methods for producing, titering, and qualitating, via infectivity assay, native virus produced from organotypic raft culture.

Key words

HPV Native Keratinocyte Organotypic raft culture Virion Differentiation 

References

  1. 1.
    Meyers C, Frattini MG, Hudson JB, Laimins LA (1992) Biosynthesis of human papillomavirus from a continuous cell line upon epithelial differentiation. Science 257:971–973PubMedCrossRefGoogle Scholar
  2. 2.
    Meyers C, Mayer TJ, Ozbun MA (1997) Synthesis of infectious human papillomavirus type 18 in differentiating epithelium transfected with viral DNA. J Virol 71:7381–7386PubMedPubMedCentralGoogle Scholar
  3. 3.
    Conway MJ, Alam S, Ryndock EJ et al (2009) Tissue-spanning redox gradient-dependant assembly of native human papillomavirus type 16 virions. J Virol 83:10515–10526PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Jennifer Biryukov
    • 1
  • Linda Cruz
    • 1
  • Eric J. Ryndock
    • 1
  • Craig Meyers
    • 1
  1. 1.Department of Microbiology and Immunology, College of MedicineThe Pennsylvania State UniversityHersheyUSA

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