ArticleFrontiers in immunology2022
Harmonization and qualification of an IFN-γ Enzyme-Linked ImmunoSpot assay (ELISPOT) to measure influenza-specific cell-mediated immunity within the FLUCOP consortium.
Article in Frontiers in immunology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed.
- Immune responses in healthy adults elicited by a bivalent norovirus vaccine candidate composed of GI.4 and GII.4 VLPs without adjuvant.Frontiers in immunology · 2023Trial
- T cell immunomonitoring: a comparative analysis of traditional and novel methods to quantify and characterize human antigen-specific T cells.Nature protocols · 2026Review
- Cross-reactive antibody and T-cell responses after influenza virus infection in community-dwelling older adults.Journal of virology · 2026Article
- The nanoemulsion adjuvant provides antigen dose-sparing effects and enhances maternal passive immune protection for the cell-cultured quadrivalent influenza virus subunit vaccine.Virology journal · 2026Article
- Harnessing alternative splicing for off-the-shelf mRNA neoantigen vaccines in hepatocellular carcinoma.Cell research · 2025Article
- RBD-displaying OMV nanovaccine boosts immunity against SARS-CoV-2.Journal of nanobiotechnology · 2025Article
- Editorial: Community series in development and harmonization of assays and models to assess immunogenicity and correlates of protection of vaccines against pathogens causing respiratory infections, volume 2.Frontiers in immunology · 2025Article
- T-Cell Interferon Gamma Responses to SARS-CoV-2 Following Infection with/Without Vaccination in a Tanzanian Population.Infection and drug resistance · 2025Article
- Validation of an interferon-gamma enzyme-linked immunosorbent spot assay to evaluate cell-mediated immunity against severe acute respiratory syndrome coronavirus 2.Human vaccines & immunotherapeutics · 2024Article
- Inno4Vac Workshop Report Part 1: Controlled Human Influenza Virus Infection Model (CHIVIM) Strain Selection and Immune Assays for CHIVIM Studies, November 2021, MHRA, UK.Influenza and other respiratory viruses · 2024Review
- Article
- Meeting Report From "Correlates of Protection for Next Generation Influenza Vaccines: Lessons Learned From the COVID-19 Pandemic".Influenza and other respiratory viruses · 2024Article
- Meeting Report: Controlled Human Influenza Virus Infection Model Studies: Current Status and Future Directions for Innovation.Influenza and other respiratory viruses · 2024Article
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- SARS-CoV-2 epitope-specific T cells: Immunity response feature, TCR repertoire characteristics and cross-reactivity.Frontiers in immunology · 2023Review
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13 authors.
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Abstract
Influenza continues to be the most important cause of viral respiratory disease, despite the availability of vaccines. Today's evaluation of influenza vaccines mainly focuses on the quantitative and functional analyses of antibodies to the surface proteins haemagglutinin (HA) and neuraminidase (NA). However, there is an increasing interest in measuring cellular immune responses targeting not only mutation-prone surface HA and NA but also conserved internal proteins as these are less explored yet potential correlates of protection. To date, laboratories that monitor cellular immune responses use a variety of in-house procedures. This generates diverging results, complicates interlaboratory comparisons, and hampers influenza vaccine evaluation. The European FLUCOP project aims to develop and standardize assays for the assessment of influenza vaccine correlates of protection. This report describes the harmonization and qualification of the influenza-specific interferon-gamma (IFN-γ) Enzyme-Linked ImmunoSpot (ELISpot) assay. Initially, two pilot studies were conducted to identify sources of variability during sample analysis and spot enumeration in order to develop a harmonized Standard Operating Procedure (SOP). Subsequently, an assay qualification study was performed to investigate the linearity, intermediate precision (reproducibility), repeatability, specificity, Lower and Upper Limits of Quantification (LLOQ-ULOQ), Limit of Detection (LOD) and the stability of signal over time. We were able to demonstrate that the FLUCOP harmonized IFN-γ ELISpot assay procedure can accurately enumerate IFN-γ secreting cells in the analytical range of 34.4 Spot Forming Units (SFU) per million cells up to the technical limit of the used reader and in the linear range from 120 000 to 360 000 cells per well, in plates stored up to 6 weeks after development. This IFN-γ ELISpot procedure will hopefully become a useful and reliable tool to investigate influenza-specific cellular immune responses induced by natural infection or vaccination and can be an additional instrument in the search for novel correlates of protection.
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