Detection of human papillomavirus (HPV) L1 gene DNA possibly bound to particulate aluminum adjuvant in the HPV vaccine Gardasil

J Inorg Biochem. 2012 Dec:117:85-92. doi: 10.1016/j.jinorgbio.2012.08.015. Epub 2012 Aug 30.

Abstract

Medical practitioners in nine countries submitted samples of Gardasil (Merck & Co.) to be tested for the presence of human papillomavirus (HPV) DNA because they suspected that residual recombinant HPV DNA left in the vaccine might have been a contributing factor leading to some of the unexplained post-vaccination side effects. A total of 16 packages of Gardasil were received from Australia, Bulgaria, France, India, New Zealand, Poland, Russia, Spain and the United States. A nested polymerase chain reaction (PCR) method using the MY09/MY11 degenerate primers for initial amplification and the GP5/GP6-based nested PCR primers for the second amplification were used to prepare the template for direct automated cycle DNA sequencing of a hypervariable segment of the HPV L1 gene which is used for manufacturing of the HPV L1 capsid protein by a DNA recombinant technology in vaccine production. Detection of HPV DNA and HPV genotyping of all positive samples were finally validated by BLAST (Basic Local Alignment Search Tool) analysis of a 45-60 bases sequence of the computer-generated electropherogram. The results showed that all 16 Gardasil samples, each with a different lot number, contained fragments of HPV-11 DNA, or HPV-18 DNA, or a DNA fragment mixture from both genotypes. The detected HPV DNA was found to be firmly bound to the insoluble, proteinase-resistant fraction, presumably of amorphous aluminum hydroxyphosphate sulfate (AAHS) nanoparticles used as adjuvant. The clinical significance of these residual HPV DNA fragments bound to a particulate mineral-based adjuvant is uncertain after intramuscular injection, and requires further investigation for vaccination safety.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Pharmaceutic / chemistry*
  • Aluminum Hydroxide / chemistry
  • Base Sequence
  • Capsid Proteins / genetics*
  • DNA Primers / chemistry
  • DNA, Viral / chemistry*
  • DNA, Viral / genetics
  • Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18
  • Human papillomavirus 11 / genetics*
  • Human papillomavirus 18 / genetics*
  • Humans
  • Molecular Sequence Data
  • Oncogene Proteins, Viral / genetics*
  • Papillomavirus Vaccines / chemistry*
  • Papillomavirus Vaccines / genetics
  • Phosphates / chemistry
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA

Substances

  • Adjuvants, Pharmaceutic
  • Capsid Proteins
  • DNA Primers
  • DNA, Viral
  • HPV L1 protein, Human papillomavirus
  • Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18
  • Oncogene Proteins, Viral
  • Papillomavirus Vaccines
  • Phosphates
  • Aluminum Hydroxide
  • aluminum hydroxyphosphate sulfate