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An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses

Numerous assays evaluating serological and cellular responses have been developed to characterize immune responses against SARS-CoV-2. Serological assays are both cost- and time-effective compared to cellular assays, but cellular immune responses may provide a diagnostic value to determine previous...

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Autores principales: Mangsbo, Sara M., Havervall, Sebastian, Laurén, Ida, Lindsay, Robin, Jernbom Falk, August, Marking, Ulrika, Lord, Martin, Buggert, Marcus, Dönnes, Pierre, Christoffersson, Gustaf, Nilsson, Peter, Hober, Sophia, Phillipson, Mia, Klingström, Jonas, Thålin, Charlotte
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8483319/
https://www.ncbi.nlm.nih.gov/pubmed/34591918
http://dx.doi.org/10.1371/journal.pone.0258041
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author Mangsbo, Sara M.
Havervall, Sebastian
Laurén, Ida
Lindsay, Robin
Jernbom Falk, August
Marking, Ulrika
Lord, Martin
Buggert, Marcus
Dönnes, Pierre
Christoffersson, Gustaf
Nilsson, Peter
Hober, Sophia
Phillipson, Mia
Klingström, Jonas
Thålin, Charlotte
author_facet Mangsbo, Sara M.
Havervall, Sebastian
Laurén, Ida
Lindsay, Robin
Jernbom Falk, August
Marking, Ulrika
Lord, Martin
Buggert, Marcus
Dönnes, Pierre
Christoffersson, Gustaf
Nilsson, Peter
Hober, Sophia
Phillipson, Mia
Klingström, Jonas
Thålin, Charlotte
author_sort Mangsbo, Sara M.
collection PubMed
description Numerous assays evaluating serological and cellular responses have been developed to characterize immune responses against SARS-CoV-2. Serological assays are both cost- and time-effective compared to cellular assays, but cellular immune responses may provide a diagnostic value to determine previous SARS-CoV-2 infection in seronegative individuals. However, potential cross-reactive T cell responses stemming from prior encounters with human coronaviruses (HCoVs) may affect assay specificity. In this study, we evaluated the specificity and sensitivity of a SARS-CoV-2 IFN-γ Release Assay (IGRA) based on the FluoroSpot method employing commercially available SARS-CoV-2-specific peptide pools, as well as an in-house designed SARS-CoV-2 peptide pool restricted to 5 amino acid stretches or less aligning with endemic HCoVs. Blood samples were obtained from healthcare workers (HCW) 5–6 months post SARS-CoV-2 spike (S) IgG and nucleocapsid (N) IgG dual seroconversion (n = 187) and HCW who had been S IgG and N IgG dual seronegative at repeated occasions, including the current sampling time point (n = 102). In addition, samples were obtained 4 to 5 months post infection from 55 polymerase chain reaction (PCR)-confirmed COVID-19 patients. Assay specificity and sensitivity were calculated with serology as a reference standard for HCW. The in-house generated peptide pool displayed a specificity of 96.1%, while the commercially available peptide pools displayed specificities of 80.4% and 85.3%, respectively. Sensitivity was higher in a cohort of previously hospitalized COVID-19 patients (96.4% and 84.0% for the commercially available peptide pools and 92.7% for the in-house generated peptide pool) compared to the HCW cohort (92.0% and 66.8% for the commercially available peptide pools and 76.0% for the in-house generated peptide pool). Based on these findings, the individual diagnostic value of T cell immune responses against SARS-CoV-2 currently appears to be limited but remain an important research tool ahead.
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spelling pubmed-84833192021-10-01 An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses Mangsbo, Sara M. Havervall, Sebastian Laurén, Ida Lindsay, Robin Jernbom Falk, August Marking, Ulrika Lord, Martin Buggert, Marcus Dönnes, Pierre Christoffersson, Gustaf Nilsson, Peter Hober, Sophia Phillipson, Mia Klingström, Jonas Thålin, Charlotte PLoS One Research Article Numerous assays evaluating serological and cellular responses have been developed to characterize immune responses against SARS-CoV-2. Serological assays are both cost- and time-effective compared to cellular assays, but cellular immune responses may provide a diagnostic value to determine previous SARS-CoV-2 infection in seronegative individuals. However, potential cross-reactive T cell responses stemming from prior encounters with human coronaviruses (HCoVs) may affect assay specificity. In this study, we evaluated the specificity and sensitivity of a SARS-CoV-2 IFN-γ Release Assay (IGRA) based on the FluoroSpot method employing commercially available SARS-CoV-2-specific peptide pools, as well as an in-house designed SARS-CoV-2 peptide pool restricted to 5 amino acid stretches or less aligning with endemic HCoVs. Blood samples were obtained from healthcare workers (HCW) 5–6 months post SARS-CoV-2 spike (S) IgG and nucleocapsid (N) IgG dual seroconversion (n = 187) and HCW who had been S IgG and N IgG dual seronegative at repeated occasions, including the current sampling time point (n = 102). In addition, samples were obtained 4 to 5 months post infection from 55 polymerase chain reaction (PCR)-confirmed COVID-19 patients. Assay specificity and sensitivity were calculated with serology as a reference standard for HCW. The in-house generated peptide pool displayed a specificity of 96.1%, while the commercially available peptide pools displayed specificities of 80.4% and 85.3%, respectively. Sensitivity was higher in a cohort of previously hospitalized COVID-19 patients (96.4% and 84.0% for the commercially available peptide pools and 92.7% for the in-house generated peptide pool) compared to the HCW cohort (92.0% and 66.8% for the commercially available peptide pools and 76.0% for the in-house generated peptide pool). Based on these findings, the individual diagnostic value of T cell immune responses against SARS-CoV-2 currently appears to be limited but remain an important research tool ahead. Public Library of Science 2021-09-30 /pmc/articles/PMC8483319/ /pubmed/34591918 http://dx.doi.org/10.1371/journal.pone.0258041 Text en © 2021 Mangsbo et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Mangsbo, Sara M.
Havervall, Sebastian
Laurén, Ida
Lindsay, Robin
Jernbom Falk, August
Marking, Ulrika
Lord, Martin
Buggert, Marcus
Dönnes, Pierre
Christoffersson, Gustaf
Nilsson, Peter
Hober, Sophia
Phillipson, Mia
Klingström, Jonas
Thålin, Charlotte
An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title_full An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title_fullStr An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title_full_unstemmed An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title_short An evaluation of a FluoroSpot assay as a diagnostic tool to determine SARS-CoV-2 specific T cell responses
title_sort evaluation of a fluorospot assay as a diagnostic tool to determine sars-cov-2 specific t cell responses
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8483319/
https://www.ncbi.nlm.nih.gov/pubmed/34591918
http://dx.doi.org/10.1371/journal.pone.0258041
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