{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T05:23:15Z","timestamp":1773984195736,"version":"3.50.1"},"reference-count":43,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,12]],"date-time":"2022-09-12T00:00:00Z","timestamp":1662940800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001349","name":"National Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001349","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001349","name":"National Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001349","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Increased neutrophils and elevated level of circulating calprotectin are hallmarks of severe COVID-19 and they contribute to the dysregulated immune responses and cytokine storm in susceptible patients. However, the precise mechanism controlling calprotectin production during SARS-CoV-2 infection remains elusive. In this study, we showed that Dok3 adaptor restrains calprotectin production by neutrophils in response to SARS-CoV-2 spike (S) protein engagement of TLR4. Dok3 recruits SHP-2 to mediate the de-phosphorylation of MyD88 at Y257, thereby attenuating downstream JAK2-STAT3 signaling and calprotectin production. Blocking of TLR4, JAK2 and STAT3 signaling could prevent excessive production of calprotectin by Dok3<jats:sup>-\/-<\/jats:sup> neutrophils, revealing new targets for potential COVID-19 therapy. As S protein from SARS-CoV-2 Delta and Omicron variants can activate TLR4-driven calprotectin production in Dok3<jats:sup>-\/-<\/jats:sup> neutrophils, our study suggests that targeting calprotectin production may be an effective strategy to combat severe COVID-19 manifestations associated with these emerging variants.<\/jats:p>","DOI":"10.3389\/fimmu.2022.996637","type":"journal-article","created":{"date-parts":[[2022,9,12]],"date-time":"2022-09-12T06:18:02Z","timestamp":1662963482000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Dok3 restrains neutrophil production of calprotectin during TLR4 sensing of SARS-CoV-2 spike protein"],"prefix":"10.3389","volume":"13","author":[{"given":"Jia Tong","family":"Loh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joey Kay Hui","family":"Teo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kong-Peng","family":"Lam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,12]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.652470","article-title":"Neutrophils in COVID-19","volume":"12","author":"Reusch","year":"2021","journal-title":"Front Immunol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2116597","article-title":"Effect of covid-19 vaccination on transmission of alpha and delta variants","volume":"386","author":"Eyre","year":"2022","journal-title":"New Engl J Med"},{"key":"B3","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1038\/s41577-021-00662-4","article-title":"SARS-CoV-2 breakthrough infections in vaccinated individuals: measurement, causes and impact","volume":"22","author":"Lipsitch","year":"2022","journal-title":"Nat Rev Immunol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.1c01451","article-title":"Omicron variant (B.1.1.529): Infectivity, vaccine breakthrough, and antibody resistance","volume":"62","author":"Chen","year":"2022","journal-title":"J Chem Inf Model"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-04594-4","article-title":"Antibody evasion properties of SARS-CoV-2 omicron sublineages","volume":"604","author":"Iketani","year":"2022","journal-title":"Nature"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-021-01678-y","article-title":"An infectious SARS-CoV-2 B.1.1.529 omicron virus escapes neutralization by therapeutic monoclonal antibodies","volume":"28","author":"VanBlargan","year":"2022","journal-title":"Nat Med"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cytogfr.2021.10.004","article-title":"S100A8\/A9 in COVID-19 pathogenesis: Impact on clinical outcomes","volume":"63","author":"Mellett","year":"2022","journal-title":"Cytokine Growth Factor Rev"},{"key":"B8","doi-asserted-by":"publisher","first-page":"1401","DOI":"10.1016\/j.cell.2020.08.002","article-title":"Elevated calprotectin and abnormal myeloid cell subsets discriminate severe from mild COVID-19","volume":"182","author":"Silvin","year":"2020","journal-title":"Cell"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1080\/1744666X.2021.1905526","article-title":"Circulating calprotectin as a biomarker of COVID-19 severity","volume":"17","author":"Mahler","year":"2021","journal-title":"Expert Rev Clin Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-020-0492-x","article-title":"Elevated serum levels of S100A8\/A9 and HMGB1 at hospital admission are correlated with inferior clinical outcomes in COVID-19 patients","volume":"17","author":"Chen","year":"2020","journal-title":"Cell Mol Immunol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.751232","article-title":"Elevated levels of neutrophil activated proteins, alpha-defensins (DEFA1), calprotectin (S100A8\/A9) and myeloperoxidase (MPO) are associated with disease severity in COVID-19 patients","volume":"11","author":"Shrivastava","year":"2021","journal-title":"Front Cell Infect Microbiol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.01298","article-title":"S100A8\/A9 in inflammation","volume":"9","author":"Wang","year":"2018","journal-title":"Front Immunol"},{"key":"B13","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1016\/j.chom.2020.12.016","article-title":"Induction of alarmin S100A8\/A9 mediates activation of aberrant neutrophils in the pathogenesis of COVID-19","volume":"29","author":"Guo","year":"2021","journal-title":"Cell Host Microbe"},{"key":"B14","doi-asserted-by":"publisher","first-page":"e21798","DOI":"10.1096\/fj.202101013","article-title":"S100A8 may govern hyper-inflammation in severe COVID-19","volume":"35","author":"Deguchi","year":"2021","journal-title":"FASEB J"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.566192","article-title":"Emerging roles of downstream of kinase 3 in cell signaling","volume":"11","author":"Loh","year":"2020","journal-title":"Front Immunol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1172\/JCI126341","article-title":"Dok3-protein phosphatase 1 interaction attenuates Card9 signaling and neutrophil-dependent antifungal immunity","volume":"129","author":"Loh","year":"2019","journal-title":"J Clin Invest"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1186\/s13073-020-00823-5","article-title":"Disease severity-specific neutrophil signatures in blood transcriptomes stratify COVID-19 patients","volume":"13","author":"Aschenbrenner","year":"2021","journal-title":"Genome Med"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1038\/s41422-021-00495-9","article-title":"SARS-CoV-2 spike protein interacts with and activates TLR41","volume":"31","author":"Zhao","year":"2021","journal-title":"Cell Res"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.68563","article-title":"SARS-CoV-2 spike protein induces inflammation via TLR2-dependent activation of the NF-\u03baB pathway","volume":"10","author":"Khan","year":"2021","journal-title":"Elife"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2021.e06187","article-title":"SARS-CoV-2 spike protein S1 subunit induces pro-inflammatory responses via toll-like receptor 4 signaling in murine and human macrophages","volume":"7","author":"Shirato","year":"2021","journal-title":"Heliyon"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2180-5","article-title":"Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor","volume":"581","author":"Lan","year":"2020","journal-title":"Nature"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0039967","article-title":"DOK3 negatively regulates LPS responses and endotoxin tolerance","volume":"7","author":"Peng","year":"2012","journal-title":"PloS One"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1038\/s41366-021-00768-8","article-title":"Increased TLR\/MyD88 signaling in patients with obesity: is there a link to COVID-19 disease severity","volume":"45","author":"Cuevas","year":"2021","journal-title":"Int J Obes"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-00937-x","article-title":"TLR2 senses the SARS-CoV-2 envelope protein to produce inflammatory cytokines","volume":"22","author":"Zheng","year":"2021","journal-title":"Nat Immunol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2017.03.014","article-title":"Tyrosine kinase SYK licenses MyD88 adaptor protein to instigate IL-1\u03b1-Mediated inflammatory disease","volume":"46","author":"Gurung","year":"2017","journal-title":"Immunity"},{"key":"B26","doi-asserted-by":"publisher","first-page":"1054","DOI":"10.1038\/s41419-021-04357-5","article-title":"DOK3 maintains intestinal homeostasis by suppressing JAK2\/STAT3 signaling and S100a8\/9 production in neutrophils","volume":"12","author":"Loh","year":"2021","journal-title":"Cell Death Dis"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0508517102","article-title":"Suppressor of cytokine signaling-1 selectively inhibits LPS-induced IL-6 production by regulating JAK\u2013STAT","volume":"102","author":"Akihiro","year":"2005","journal-title":"Proc Natl Acad Sci"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2006.08.023","article-title":"The adaptor molecule MyD88 activates PI-3 kinase signaling in CD4+ T cells and enables CpG oligodeoxynucleotide-mediated costimulation","volume":"25","author":"Gelman","year":"2006","journal-title":"Immunity"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1038\/s41375-021-01266-6","article-title":"JAK-inhibitors for coronavirus disease-2019 (COVID-19): a meta-analysis","volume":"35","author":"Chen","year":"2021","journal-title":"Leukemia"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3389\/fmed.2021.800492","article-title":"The efficacy and safety of janus kinase inhibitors for patients with COVID-19: A living systematic review and meta-analysis","volume":"8","author":"Zhang","year":"2022","journal-title":"Front Med"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1245","DOI":"10.1136\/annrheumdis-2021-220049","article-title":"Use of janus kinase inhibitors in COVID-19: a prospective observational series in 522 individuals","volume":"80","author":"Melikhov","year":"2021","journal-title":"Ann Rheum Dis"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-021-00805-z","article-title":"Patients with COVID-19: in the dark-NETs of neutrophils","volume":"28","author":"Ackermann","year":"2021","journal-title":"Cell Death Differ"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2021.708302","article-title":"NETosis and the immune system in COVID-19: Mechanisms and potential treatments","volume":"12","author":"Gillot","year":"2021","journal-title":"Front Pharmacol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.138999","article-title":"Neutrophil extracellular traps (NETs) as markers of disease severity in COVID-19","volume":"5","author":"Zuo","year":"2020","journal-title":"JCI Insight"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20201129","article-title":"SARS-CoV-2\u2013triggered neutrophil extracellular traps mediate COVID-19 pathologySARS-CoV-2 directly triggers ACE-dependent NETs","volume":"217","author":"Veras","year":"2020","journal-title":"J Exp Med"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2021.103406","article-title":"A lethal mouse model using a mouse-adapted SARS-CoV-2 strain with enhanced binding to mouse ACE2 as an important platform for COVID-19 research","volume":"68","author":"Chu","year":"2021","journal-title":"EBioMedicine"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-020-0778-2","article-title":"SARS-CoV-2 infection of human ACE2-transgenic mice causes severe lung inflammation and impaired function","volume":"21","author":"Winkler","year":"2020","journal-title":"Nat Immunol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2708-8","article-title":"A mouse-adapted model of SARS-CoV-2 to test COVID-19 countermeasures","volume":"586","author":"Dinnon","year":"2020","journal-title":"Nature"},{"key":"B39","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1001\/jamainternmed.2020.6820","article-title":"Group c-19 c. effect of tocilizumab vs usual care in adults hospitalized with COVID-19 and moderate or severe pneumonia: A randomized clinical trial","volume":"181","author":"Hermine","year":"2021","journal-title":"JAMA Internal Med"},{"key":"B40","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1001\/jamainternmed.2020.6615","article-title":"Effect of tocilizumab vs standard care on clinical worsening in patients hospitalized with COVID-19 pneumonia: A randomized clinical trial","volume":"181","author":"Salvarani","year":"2021","journal-title":"JAMA Internal Med"},{"key":"B41","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1056\/NEJMoa2030340","article-title":"Tocilizumab in patients hospitalized with covid-19 pneumonia","volume":"384","author":"Salama","year":"2020","journal-title":"New Engl J Med"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2028836","article-title":"Efficacy of tocilizumab in patients hospitalized with covid-19","volume":"383","author":"Stone","year":"2020","journal-title":"New Engl J Med"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-10-055194","article-title":"Dok-3 plays a nonredundant role in negative regulation of b-cell activation","volume":"110","author":"Ng","year":"2007","journal-title":"Blood"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.996637\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,9,12]],"date-time":"2022-09-12T06:18:05Z","timestamp":1662963485000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.996637\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,12]]},"references-count":43,"alternative-id":["10.3389\/fimmu.2022.996637"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.996637","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,12]]},"article-number":"996637"}}