Spike is the most recognized antigen in the whole-blood platform in both acute and convalescent COVID-19 patients

•Whole-blood interferon-γ-release assay (IGRA) enabled the response to SARS-CoV-2 peptides to be evaluated.•Spike protein was the best stimulus to discriminate COVID-19 from NO-COVID-19.•A SARS-CoV-2-specific response is rarely observed in NO-COVID-19 individuals.•This SARS-CoV-2 IGRA has the potent...

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Published in:International journal of infectious diseases Vol. 106; pp. 338 - 347
Main Authors: Aiello, Alessandra, Najafi Fard, Saeid, Petruccioli, Elisa, Petrone, Linda, Vanini, Valentina, Farroni, Chiara, Cuzzi, Gilda, Navarra, Assunta, Gualano, Gina, Mosti, Silvia, Pierelli, Luca, Nicastri, Emanuele, Goletti, Delia
Format: Journal Article
Language:English
Published: Canada Elsevier Ltd 01.05.2021
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ISSN:1201-9712, 1878-3511, 1878-3511
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Abstract •Whole-blood interferon-γ-release assay (IGRA) enabled the response to SARS-CoV-2 peptides to be evaluated.•Spike protein was the best stimulus to discriminate COVID-19 from NO-COVID-19.•A SARS-CoV-2-specific response is rarely observed in NO-COVID-19 individuals.•This SARS-CoV-2 IGRA has the potential to be a powerful diagnostic tool. To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Whole-blood from 56 COVID-19 and 23 “NO-COVID-19” individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels. The IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14–2.17; and median: 1.18, IQR: 0.27–4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032–1.26; and median: 0.22, IQR: 0.01−0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4+ and CD8+ T cells, and independently detected of clinical parameters in both hospitalized and recovered patients. This easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.
AbstractList To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform.OBJECTIVESTo identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform.Whole-blood from 56 COVID-19 and 23 "NO-COVID-19" individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels.METHODSWhole-blood from 56 COVID-19 and 23 "NO-COVID-19" individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels.The IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14-2.17; and median: 1.18, IQR: 0.27-4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032-1.26; and median: 0.22, IQR: 0.01-0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4+ and CD8+ T cells, and independently detected of clinical parameters in both hospitalized and recovered patients.RESULTSThe IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14-2.17; and median: 1.18, IQR: 0.27-4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032-1.26; and median: 0.22, IQR: 0.01-0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4+ and CD8+ T cells, and independently detected of clinical parameters in both hospitalized and recovered patients.This easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.CONCLUSIONSThis easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.
•Whole-blood interferon-γ-release assay (IGRA) enabled the response to SARS-CoV-2 peptides to be evaluated.•Spike protein was the best stimulus to discriminate COVID-19 from NO-COVID-19.•A SARS-CoV-2-specific response is rarely observed in NO-COVID-19 individuals.•This SARS-CoV-2 IGRA has the potential to be a powerful diagnostic tool. To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Whole-blood from 56 COVID-19 and 23 “NO-COVID-19” individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels. The IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14–2.17; and median: 1.18, IQR: 0.27–4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032–1.26; and median: 0.22, IQR: 0.01−0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4+ and CD8+ T cells, and independently detected of clinical parameters in both hospitalized and recovered patients. This easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.
Objectives: To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Methods: Whole-blood from 56 COVID-19 and 23 “NO-COVID-19” individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels. Results: The IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14–2.17; and median: 1.18, IQR: 0.27–4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032–1.26; and median: 0.22, IQR: 0.01−0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4+ and CD8+ T cells, and independently detected of clinical parameters in both hospitalized and recovered patients. Conclusions: This easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.
To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Whole-blood from 56 COVID-19 and 23 "NO-COVID-19" individuals were stimulated overnight with different concentrations (0.1 or 1 μg/mL) of SARS-CoV-2 PepTivator® Peptide Pools, including spike (pool S), nucleocapsid (pool N), membrane (pool M), and a MegaPool (MP) of these three peptide pools. ELISA was used to analyse interferon (IFN)-γ levels. The IFN-γ-response to every SARS-CoV-2 peptide pool was significantly increased in COVID-19 patients compared with NO-COVID-19 individuals. Pool S and MegaPool were the most potent immunogenic stimuli (median: 0.51, IQR: 0.14-2.17; and median: 1.18, IQR: 0.27-4.72, respectively) compared with pools N and M (median: 0.22, IQR: 0.032-1.26; and median: 0.22, IQR: 0.01-0.71, respectively). The whole-blood test based on pool S and MegaPool showed a good sensitivity of 77% and a high specificity of 96%. The IFN-γ-response was mediated by both CD4 and CD8 T cells, and independently detected of clinical parameters in both hospitalized and recovered patients. This easy-to-use assay for detecting SARS-CoV-2-specific T cell responses may be implemented in clinical laboratories as a powerful diagnostic tool.
Author Nicastri, Emanuele
Petruccioli, Elisa
Farroni, Chiara
Mosti, Silvia
Aiello, Alessandra
Navarra, Assunta
Vanini, Valentina
Gualano, Gina
Goletti, Delia
Pierelli, Luca
Najafi Fard, Saeid
Petrone, Linda
Cuzzi, Gilda
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  givenname: Saeid
  surname: Najafi Fard
  fullname: Najafi Fard, Saeid
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Elisa
  surname: Petruccioli
  fullname: Petruccioli, Elisa
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Linda
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  fullname: Petrone, Linda
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Valentina
  surname: Vanini
  fullname: Vanini, Valentina
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Chiara
  surname: Farroni
  fullname: Farroni, Chiara
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Gilda
  surname: Cuzzi
  fullname: Cuzzi, Gilda
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
– sequence: 8
  givenname: Assunta
  surname: Navarra
  fullname: Navarra, Assunta
  organization: Clinical Epidemiology Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
– sequence: 9
  givenname: Gina
  surname: Gualano
  fullname: Gualano, Gina
  organization: Clinical Division of Respiratory Infectious Diseases, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
– sequence: 10
  givenname: Silvia
  surname: Mosti
  fullname: Mosti, Silvia
  organization: Clinical Division of Respiratory Infectious Diseases, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
– sequence: 11
  givenname: Luca
  surname: Pierelli
  fullname: Pierelli, Luca
  organization: UOC Transfusion Medicine and Stem Cell Unit, San Camillo Forlanini Hospital, Rome, Italy
– sequence: 12
  givenname: Emanuele
  surname: Nicastri
  fullname: Nicastri, Emanuele
  organization: Clinical Division of Infectious Diseases, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
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  givenname: Delia
  orcidid: 0000-0001-8360-4376
  surname: Goletti
  fullname: Goletti, Delia
  email: delia.goletti@inmi.it
  organization: Translational Research Unit, National Institute for Infectious Diseases Lazzaro Spallanzani-IRCCS, Rome, Italy
BackLink https://www.ncbi.nlm.nih.gov/pubmed/33864921$$D View this record in MEDLINE/PubMed
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Keywords COVID-19
IFN-γ-release assay (IGRA)
SARS-CoV-2
Membrane protein
Whole-blood
Spike protein
T cell response
Nucleocapsid protein
Language English
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Snippet •Whole-blood interferon-γ-release assay (IGRA) enabled the response to SARS-CoV-2 peptides to be evaluated.•Spike protein was the best stimulus to discriminate...
To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Whole-blood from 56 COVID-19 and 23...
To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform.OBJECTIVESTo identify the best...
Objectives: To identify the best experimental approach to detect a SARS-CoV-2-specific T cell response using a whole-blood platform. Methods: Whole-blood from...
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StartPage 338
SubjectTerms COVID-19
IFN-γ-release assay (IGRA)
Membrane protein
Nucleocapsid protein
SARS-CoV-2
Spike protein
T cell response
Whole-blood
Title Spike is the most recognized antigen in the whole-blood platform in both acute and convalescent COVID-19 patients
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https://dx.doi.org/10.1016/j.ijid.2021.04.034
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