tetano
Editor, Senior Moderator
Clin Transl Immunology
. 2021 Apr 29;10(4):e1271.
doi: 10.1002/cti2.1271. eCollection 2021.
Kinetics of peripheral blood neutrophils in severe coronavirus disease 2019
Mieke Metzemaekers[SUP] 1 [/SUP], Seppe Cambier[SUP] 1 [/SUP], Marfa Blanter[SUP] 1 [/SUP], Jennifer Vandooren[SUP] 2 [/SUP], Ana Carolina de Carvalho[SUP] 1 3 4 [/SUP], Bert Malengier-Devlies[SUP] 2 [/SUP], Lore Vanderbeke[SUP] 5 [/SUP], Cato Jacobs[SUP] 6 [/SUP], Sofie Coenen[SUP] 7 [/SUP], Erik Martens[SUP] 2 [/SUP], No?mie P?rtner[SUP] 1 [/SUP], Lotte Vanbrabant[SUP] 1 [/SUP], Pierre Van Mol[SUP] 8 [/SUP], Yannick Van Herck[SUP] 9 [/SUP], Nathalie Van Aerde[SUP] 10 [/SUP], Greet Hermans[SUP] 10 [/SUP], Jan Gunst[SUP] 10 [/SUP], Alexandre Borin[SUP] 3 [/SUP], Bruna Toledo N Pereira[SUP] 4 [/SUP], Arilson Bernardo Dos Sp Gomes[SUP] 4 [/SUP], St?fanie Primon Muraro[SUP] 11 [/SUP], Gabriela Fabiano de Souza[SUP] 11 [/SUP], Alessandro S Farias[SUP] 12 [/SUP], Jos? Luiz Proenca-Modena[SUP] 11 12 [/SUP], Marco Aur?lio R Vinolo[SUP] 4 12 [/SUP], CONTAGIOUS Consortium; Pedro Elias Marques[SUP] 1 [/SUP], Carine Wouters[SUP] 2 13 14 [/SUP], Els Wauters[SUP] 15 [/SUP], Sofie Struyf[SUP] 1 [/SUP], Patrick Matthys[SUP] 2 [/SUP], Ghislain Opdenakker[SUP] 2 [/SUP], Rafael Elias Marques[SUP] 3 [/SUP], Joost Wauters[SUP] 6 [/SUP], Mieke Gouwy[SUP] 1 [/SUP], Paul Proost[SUP] 1 [/SUP]
Collaborators, Affiliations
Abstract
Objectives: Emerging evidence of dysregulation of the myeloid cell compartment urges investigations on neutrophil characteristics in coronavirus disease 2019 (COVID-19). We isolated neutrophils from the blood of COVID-19 patients receiving general ward care and from patients hospitalised at intensive care units (ICUs) to explore the kinetics of circulating neutrophils and factors important for neutrophil migration and activation.
Methods: Multicolour flow cytometry was exploited for the analysis of neutrophil differentiation and activation markers. Multiplex and ELISA technologies were used for the quantification of protease, protease inhibitor, chemokine and cytokine concentrations in plasma. Neutrophil polarisation responses were evaluated microscopically. Gelatinolytic and metalloproteinase activity in plasma was determined using a fluorogenic substrate. Co-culturing healthy donor neutrophils with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) allowed us to investigate viral replication in neutrophils.
Results: Upon ICU admission, patients displayed high plasma concentrations of granulocyte-colony-stimulating factor (G-CSF) and the chemokine CXCL8, accompanied by emergency myelopoiesis as illustrated by high levels of circulating CD10[SUP]-[/SUP], immature neutrophils with reduced CXCR2 and C5aR expression. Neutrophil elastase and non-metalloproteinase-derived gelatinolytic activity were increased in plasma from ICU patients. Significantly higher levels of circulating tissue inhibitor of metalloproteinase 1 (TIMP-1) in patients at ICU admission yielded decreased total MMP proteolytic activity in blood. COVID-19 neutrophils were hyper-responsive to CXCL8 and CXCL12 in shape change assays. Finally, SARS-CoV-2 failed to replicate inside human neutrophils.
Conclusion: Our study provides detailed insights into the kinetics of neutrophil phenotype and function in severe COVID-19 patients, and supports the concept of an increased neutrophil activation state in the circulation.
Keywords: COVID?19; chemokine; cytokine; emergency myelopoiesis; neutrophil; protease.
. 2021 Apr 29;10(4):e1271.
doi: 10.1002/cti2.1271. eCollection 2021.
Kinetics of peripheral blood neutrophils in severe coronavirus disease 2019
Mieke Metzemaekers[SUP] 1 [/SUP], Seppe Cambier[SUP] 1 [/SUP], Marfa Blanter[SUP] 1 [/SUP], Jennifer Vandooren[SUP] 2 [/SUP], Ana Carolina de Carvalho[SUP] 1 3 4 [/SUP], Bert Malengier-Devlies[SUP] 2 [/SUP], Lore Vanderbeke[SUP] 5 [/SUP], Cato Jacobs[SUP] 6 [/SUP], Sofie Coenen[SUP] 7 [/SUP], Erik Martens[SUP] 2 [/SUP], No?mie P?rtner[SUP] 1 [/SUP], Lotte Vanbrabant[SUP] 1 [/SUP], Pierre Van Mol[SUP] 8 [/SUP], Yannick Van Herck[SUP] 9 [/SUP], Nathalie Van Aerde[SUP] 10 [/SUP], Greet Hermans[SUP] 10 [/SUP], Jan Gunst[SUP] 10 [/SUP], Alexandre Borin[SUP] 3 [/SUP], Bruna Toledo N Pereira[SUP] 4 [/SUP], Arilson Bernardo Dos Sp Gomes[SUP] 4 [/SUP], St?fanie Primon Muraro[SUP] 11 [/SUP], Gabriela Fabiano de Souza[SUP] 11 [/SUP], Alessandro S Farias[SUP] 12 [/SUP], Jos? Luiz Proenca-Modena[SUP] 11 12 [/SUP], Marco Aur?lio R Vinolo[SUP] 4 12 [/SUP], CONTAGIOUS Consortium; Pedro Elias Marques[SUP] 1 [/SUP], Carine Wouters[SUP] 2 13 14 [/SUP], Els Wauters[SUP] 15 [/SUP], Sofie Struyf[SUP] 1 [/SUP], Patrick Matthys[SUP] 2 [/SUP], Ghislain Opdenakker[SUP] 2 [/SUP], Rafael Elias Marques[SUP] 3 [/SUP], Joost Wauters[SUP] 6 [/SUP], Mieke Gouwy[SUP] 1 [/SUP], Paul Proost[SUP] 1 [/SUP]
Collaborators, Affiliations
- PMID: 33968405
- PMCID: PMC8082714
- DOI: 10.1002/cti2.1271
Abstract
Objectives: Emerging evidence of dysregulation of the myeloid cell compartment urges investigations on neutrophil characteristics in coronavirus disease 2019 (COVID-19). We isolated neutrophils from the blood of COVID-19 patients receiving general ward care and from patients hospitalised at intensive care units (ICUs) to explore the kinetics of circulating neutrophils and factors important for neutrophil migration and activation.
Methods: Multicolour flow cytometry was exploited for the analysis of neutrophil differentiation and activation markers. Multiplex and ELISA technologies were used for the quantification of protease, protease inhibitor, chemokine and cytokine concentrations in plasma. Neutrophil polarisation responses were evaluated microscopically. Gelatinolytic and metalloproteinase activity in plasma was determined using a fluorogenic substrate. Co-culturing healthy donor neutrophils with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) allowed us to investigate viral replication in neutrophils.
Results: Upon ICU admission, patients displayed high plasma concentrations of granulocyte-colony-stimulating factor (G-CSF) and the chemokine CXCL8, accompanied by emergency myelopoiesis as illustrated by high levels of circulating CD10[SUP]-[/SUP], immature neutrophils with reduced CXCR2 and C5aR expression. Neutrophil elastase and non-metalloproteinase-derived gelatinolytic activity were increased in plasma from ICU patients. Significantly higher levels of circulating tissue inhibitor of metalloproteinase 1 (TIMP-1) in patients at ICU admission yielded decreased total MMP proteolytic activity in blood. COVID-19 neutrophils were hyper-responsive to CXCL8 and CXCL12 in shape change assays. Finally, SARS-CoV-2 failed to replicate inside human neutrophils.
Conclusion: Our study provides detailed insights into the kinetics of neutrophil phenotype and function in severe COVID-19 patients, and supports the concept of an increased neutrophil activation state in the circulation.
Keywords: COVID?19; chemokine; cytokine; emergency myelopoiesis; neutrophil; protease.