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Optimizing Microneutralization and IFN-γ ELISPOT Assays to Evaluate Mpox Immunity.

Yinyi Yu1, Krystal Meza1, Chase Colbert1

  • 1Division of Infectious Diseases, Allergy and Immunology, Saint Louis University, St. Louis, MO 63104, USA.

Vaccines
|January 24, 2025
PubMed
Summary

New assays for mpox (monkeypox virus) neutralizing antibodies and T cell responses were developed. These robust assays show promise for future mpox vaccine efficacy studies.

Keywords:
FRNTIFN-γ ELISPOTPRNTmicroneutralizationmpoxvaccinia

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Area of Science:

  • Immunology
  • Virology
  • Vaccinology

Background:

  • Current pox virus assays are time-consuming and may require specific antigens.
  • There is a need for efficient methods to measure immune responses to mpox (monkeypox virus).

Purpose of the Study:

  • To develop and validate robust assays for measuring mpox-specific neutralizing antibodies.
  • To develop and validate robust assays for measuring mpox-specific T cell responses.

Main Methods:

  • Focused Reduction Neutralization Test (FRNT) was optimized and compared to plaque reduction neutralization test (PRNT).
  • Interferon-gamma (IFN-γ) ELISPOT assay was optimized using UV-inactivated mpox antigen.
  • Assays were performed on samples from volunteers vaccinated with Modified Vaccinia Ankara-Bavarian Nordic (MVA-BN).

Main Results:

  • The FRNT showed good agreement with PRNT (R² = 0.787).
  • Post-vaccination sera showed significantly higher neutralizing antibody titers compared to pre-vaccination sera.
  • The IFN-γ ELISPOT assay detected significantly higher T cell responses in post-vaccination samples.

Conclusions:

  • A novel microneutralization test correlates well with PRNT for mpox antibodies.
  • UV-inactivated mpox is suitable for ELISPOT assays to detect cross-reactive T cells.
  • These developed assays are valuable tools for future mpox vaccine research.