Granzyme B ELISPOT assay to measure influenza-specific cellular immunity

Hannah M. Salk, Iana H. Haralambieva, Inna G. Ovsyannikova, Krista M. Goergen, Gregory A. Poland

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


The immunogenicity and efficacy of influenza vaccination are markedly lower in the elderly. Granzyme B (GrzB), quantified in fresh cell lysates, has been suggested to be a marker of cytotoxic T lymphocyte (CTL) response and a predictor of influenza illness among vaccinated older individuals. We have developed an influenza-specific GrzB ELISPOT assay using cryopreserved PBMCs. This method was tested on 106 healthy older subjects (ages 50-74) at baseline (Day 0) and three additional time points post-vaccination (Day 3, Day 28, Day 75) with influenza A/H1N1-containing vaccine. No significant difference was seen in GrzB response between any of the time points, although influenza-specific GrzB response appears to be elevated at all post-vaccination time points. There was no correlation between GrzB response and hemagglutination inhibition (HAI) titers, indicating no relationship between the cytolytic activity and humoral antibody levels in this cohort. Additionally, a significant negative correlation between GrzB response and age was observed. These results reveal a reduction in influenza-specific GrzB response as one ages. In conclusion, we have developed and optimized an influenza-specific ELISPOT assay for use with frozen cells to quantify the CTL-specific serine protease GrzB, as a measure of cellular immunity after influenza vaccination.

Original languageEnglish (US)
Pages (from-to)44-50
Number of pages7
JournalJournal of Immunological Methods
Issue number1
StatePublished - 2013


  • Cellular
  • Cellular immunity
  • Cytotoxic
  • Cytotoxic T lymphocytes (CTL)
  • Enzyme-linked immunospot assay
  • Granzyme B
  • Granzymes
  • Human
  • Immunity
  • Influenza
  • Influenza virus
  • Killer cells
  • Natural
  • Perforin
  • T-lymphocytes
  • Viruses

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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