@article{Riou2020a,
abstract = {Rapid tests to evaluate SARS-CoV-2-specific T cell responses are urgently needed to decipher protective immunity and aid monitoring vaccine-induced immunity. Using a rapid whole blood assay requiring minimal amount of blood, we measured qualitatively and quantitatively SARS-CoV-2-specific CD4 T cell responses in 31 healthcare workers, using flow cytometry. 100{\%} of COVID-19 convalescent participants displayed a detectable SARS-CoV-2-specific CD4 T cell response. SARS-CoV-2-responding cells were also detected in 40.9{\%} of participants with no COVID-19-associated symptoms or who tested PCR negative. Phenotypic assessment indicated that, in COVID-19 convalescent participants, SARS-CoV-2 CD4 responses displayed an early differentiated memory phenotype with limited capacity to produce IFNɣ. Conversely, in participants with no reported symptoms, SARS-CoV-2 CD4 responses were enriched in late differentiated cells, co-expressing IFNɣ and TNF$\alpha$ and also Granzyme B. This proof of concept study presents a scalable alternative to PBMC-based assays to enumerate and phenotype SARS-CoV-2-responding T cells, thus representing a practical tool to monitor adaptive immunity in vaccine trials. {\#}{\#}{\#} Competing Interest Statement The authors have declared no competing interest. {\#}{\#}{\#} Funding Statement Funding sources: This work was supported by Wellcome [grant number: 104803 and 203135] and Crick idea to innovation (i2i) scheme in partnership with the Rosetrees Trust [grant number: 2020-0009]. CR is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme supported by the European Union (EU) Horizon 2020 programme [grant number: Training and Mobility Action TMA2017SF-1951-TB-SPEC to CR] and the National Institutes of Health (NIH) [grant number:R21AI148027 to CR]. GS is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme [grant number: Training and Mobility Action TMA2018SF-2446 to GS]. RJW and KAW receive support from the Francis Crick institute, which is funded by the UK Medical Research Council UKRI, Cancer Research UK and Wellcome [grant number: FC0010218]. {\#}{\#}{\#} Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The University of Cape Town Faculty of Health Sciences Human Research Ethics Committee approved the study (HREC: 207/2020) and written informed consent was obtained from all participants. All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes all data is available upon request},
author = {Riou, Catherine and Sch{\"{a}}fer, Georgia and {Du Bruyn}, Elsa and Goliath, Rene T and Stek, Cari and Hung, Deli and Wilkinson, Katalin A and Wilkinson, Robert J},
doi = {10.1101/2020.10.30.20223099},
file = {:C$\backslash$:/Users/01462563/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Riou et al. - 2020 - Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells.pdf:pdf},
journal = {medRxiv},
keywords = {OA,fund{\_}ack,original},
mendeley-tags = {OA,fund{\_}ack,original},
month = {nov},
pages = {2020.10.30.20223099},
pmid = {33173918},
publisher = {Cold Spring Harbor Laboratory Press},
title = {{Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells}},
url = {https://doi.org/10.1101/2020.10.30.20223099},
year = {2020}
}

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Phenotypic assessment indicated that, in COVID-19 convalescent participants, SARS-CoV-2 CD4 responses displayed an early differentiated memory phenotype with limited capacity to produce IFNɣ. Conversely, in participants with no reported symptoms, SARS-CoV-2 CD4 responses were enriched in late differentiated cells, co-expressing IFNɣ and TNF$α$ and also Granzyme B. This proof of concept study presents a scalable alternative to PBMC-based assays to enumerate and phenotype SARS-CoV-2-responding T cells, thus representing a practical tool to monitor adaptive immunity in vaccine trials. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement Funding sources: This work was supported by Wellcome [grant number: 104803 and 203135] and Crick idea to innovation (i2i) scheme in partnership with the Rosetrees Trust [grant number: 2020-0009]. CR is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme supported by the European Union (EU) Horizon 2020 programme [grant number: Training and Mobility Action TMA2017SF-1951-TB-SPEC to CR] and the National Institutes of Health (NIH) [grant number:R21AI148027 to CR]. GS is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme [grant number: Training and Mobility Action TMA2018SF-2446 to GS]. RJW and KAW receive support from the Francis Crick institute, which is funded by the UK Medical Research Council UKRI, Cancer Research UK and Wellcome [grant number: FC0010218]. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The University of Cape Town Faculty of Health Sciences Human Research Ethics Committee approved the study (HREC: 207/2020) and written informed consent was obtained from all participants. All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes all data is available upon request","author":[{"propositions":[],"lastnames":["Riou"],"firstnames":["Catherine"],"suffixes":[]},{"propositions":[],"lastnames":["Schäfer"],"firstnames":["Georgia"],"suffixes":[]},{"propositions":[],"lastnames":["Du Bruyn"],"firstnames":["Elsa"],"suffixes":[]},{"propositions":[],"lastnames":["Goliath"],"firstnames":["Rene","T"],"suffixes":[]},{"propositions":[],"lastnames":["Stek"],"firstnames":["Cari"],"suffixes":[]},{"propositions":[],"lastnames":["Hung"],"firstnames":["Deli"],"suffixes":[]},{"propositions":[],"lastnames":["Wilkinson"],"firstnames":["Katalin","A"],"suffixes":[]},{"propositions":[],"lastnames":["Wilkinson"],"firstnames":["Robert","J"],"suffixes":[]}],"doi":"10.1101/2020.10.30.20223099","file":":C$\\$:/Users/01462563/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Riou et al. - 2020 - Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells.pdf:pdf","journal":"medRxiv","keywords":"OA,fund_ack,original","mendeley-tags":"OA,fund_ack,original","month":"nov","pages":"2020.10.30.20223099","pmid":"33173918","publisher":"Cold Spring Harbor Laboratory Press","title":"Rapid, simplified whole blood-based multiparameter assay to quantify and phenotype SARS-CoV-2 specific T cells","url":"https://doi.org/10.1101/2020.10.30.20223099","year":"2020","bibtex":"@article{Riou2020a,\r\nabstract = {Rapid tests to evaluate SARS-CoV-2-specific T cell responses are urgently needed to decipher protective immunity and aid monitoring vaccine-induced immunity. Using a rapid whole blood assay requiring minimal amount of blood, we measured qualitatively and quantitatively SARS-CoV-2-specific CD4 T cell responses in 31 healthcare workers, using flow cytometry. 100{\\%} of COVID-19 convalescent participants displayed a detectable SARS-CoV-2-specific CD4 T cell response. SARS-CoV-2-responding cells were also detected in 40.9{\\%} of participants with no COVID-19-associated symptoms or who tested PCR negative. Phenotypic assessment indicated that, in COVID-19 convalescent participants, SARS-CoV-2 CD4 responses displayed an early differentiated memory phenotype with limited capacity to produce IFNɣ. Conversely, in participants with no reported symptoms, SARS-CoV-2 CD4 responses were enriched in late differentiated cells, co-expressing IFNɣ and TNF$\\alpha$ and also Granzyme B. This proof of concept study presents a scalable alternative to PBMC-based assays to enumerate and phenotype SARS-CoV-2-responding T cells, thus representing a practical tool to monitor adaptive immunity in vaccine trials. {\\#}{\\#}{\\#} Competing Interest Statement The authors have declared no competing interest. {\\#}{\\#}{\\#} Funding Statement Funding sources: This work was supported by Wellcome [grant number: 104803 and 203135] and Crick idea to innovation (i2i) scheme in partnership with the Rosetrees Trust [grant number: 2020-0009]. CR is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme supported by the European Union (EU) Horizon 2020 programme [grant number: Training and Mobility Action TMA2017SF-1951-TB-SPEC to CR] and the National Institutes of Health (NIH) [grant number:R21AI148027 to CR]. GS is supported by the European and Developing Countries Clinical Trials Partnership EDCTP2 programme [grant number: Training and Mobility Action TMA2018SF-2446 to GS]. RJW and KAW receive support from the Francis Crick institute, which is funded by the UK Medical Research Council UKRI, Cancer Research UK and Wellcome [grant number: FC0010218]. {\\#}{\\#}{\\#} Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The University of Cape Town Faculty of Health Sciences Human Research Ethics Committee approved the study (HREC: 207/2020) and written informed consent was obtained from all participants. All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. 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