A simple and safe antibody neutralization assay based on polio pseudoviruses

Hum Vaccin Immunother. 2019;15(2):349-357. doi: 10.1080/21645515.2018.1526553. Epub 2018 Oct 16.

Abstract

The evaluation of the immunogenicity of Sabin strain based Inactivated Poliovirus Vaccines (sIPV) necessitates the use of wild strains in neutralization assays to assess the potential cross-reactivity of antibodies. The live virus strains including wild and Sabin strains must be handled in level 3 biocontainment laboratories. To develop an alternative assay without the use of a live virus, we constructed Mahoney, MEF-1, and Saukett pseudovirions by inserting luciferase reporter genes into intact capsid proteins. Afterward, we developed a pseudovirus-based neutralization test (pNT) and evaluated for the specificity and reproducibility. We tested serum samples from a clinical trial on sIPV vaccines by pNT and compared the results with those obtained from conventional neutralization tests (cNT). A strong correlation was observed between two methods, with the correlation coefficients of all three types of IPV vaccines being greater than 0.82 (p < 0.0001). The Geometric Mean Titer (GMT) values obtained by pNT were approximately four times higher than that by cNT, revealing the better sensitivity of pNT. In conclusion, pNT is a safe, rapid and sensitive quantitative assay with the potential of being an alternative for the evaluation of the potency of polio vaccines.

Keywords: biosafety; immunogenicity evaluation; neutralization test; poliovirus vaccine; pseudovirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / blood*
  • Antibodies, Viral / blood*
  • Cell Line
  • Clinical Trials, Phase II as Topic
  • Humans
  • Neutralization Tests / methods*
  • Poliovirus / genetics
  • Poliovirus Vaccine, Inactivated / immunology*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Poliovirus Vaccine, Inactivated

Grants and funding

This work was supported by the National Science and Technology Major Projects [2012ZX10002006 and 2018ZX09737003-002].