{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T11:09:03Z","timestamp":1790420943493,"version":"4.1.0"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,26]],"date-time":"2020-08-26T00:00:00Z","timestamp":1598400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007109","name":"University of Ferrara","doi-asserted-by":"publisher","award":["COVID19 grant"],"award-info":[{"award-number":["COVID19 grant"]}],"id":[{"id":"10.13039\/501100007109","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007109","name":"University of Ferrara","doi-asserted-by":"publisher","award":["FAR 2018"],"award-info":[{"award-number":["FAR 2018"]}],"id":[{"id":"10.13039\/501100007109","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007109","name":"University of Ferrara","doi-asserted-by":"publisher","award":["FAR 2019"],"award-info":[{"award-number":["FAR 2019"]}],"id":[{"id":"10.13039\/501100007109","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Natural killer cells are important in the control of viral infections. However, the role of NK cells during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has previously not been identified. Peripheral blood NK cells from SARS-CoV and SARS-CoV-2 na\u00efve subjects were evaluated for their activation, degranulation, and interferon-gamma expression in the presence of SARS-CoV and SARS-CoV-2 spike proteins. K562 and lung epithelial cells were transfected with spike proteins and co-cultured with NK cells. The analysis was performed by flow cytometry and immune fluorescence. SARS-CoV and SARS-CoV-2 spike proteins did not alter NK cell activation in a K562 in vitro model. On the contrary, SARS-CoV-2 spike 1 protein (SP1) intracellular expression by lung epithelial cells resulted in NK cell-reduced degranulation. Further experiments revealed a concomitant induction of HLA-E expression on the surface of lung epithelial cells and the recognition of an SP1-derived HLA-E-binding peptide. Simultaneously, there was increased modulation of the inhibitory receptor NKG2A\/CD94 on NK cells when SP1 was expressed in lung epithelial cells. We ruled out the GATA3 transcription factor as being responsible for HLA-E increased levels and HLA-E\/NKG2A interaction as implicated in NK cell exhaustion. We show for the first time that NK cells are affected by SP1 expression in lung epithelial cells via HLA-E\/NKG2A interaction. The resulting NK cells\u2019 exhaustion might contribute to immunopathogenesis in SARS-CoV-2 infection.<\/jats:p>","DOI":"10.3390\/cells9091975","type":"journal-article","created":{"date-parts":[[2020,8,27]],"date-time":"2020-08-27T08:05:18Z","timestamp":1598515518000},"page":"1975","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":73,"title":["SARS-CoV-2 Spike 1 Protein Controls Natural Killer Cell Activation via the HLA-E\/NKG2A Pathway"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4496-9561","authenticated-orcid":false,"given":"Daria","family":"Bortolotti","sequence":"first","affiliation":[{"name":"Department of Chemical and Pharmaceutical Science, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4572-5497","authenticated-orcid":false,"given":"Valentina","family":"Gentili","sequence":"additional","affiliation":[{"name":"Department of Chemical and Pharmaceutical Science, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Rizzo","sequence":"additional","affiliation":[{"name":"Department of Chemical and Pharmaceutical Science, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonella","family":"Rotola","sequence":"additional","affiliation":[{"name":"Department of Chemical and Pharmaceutical Science, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9507-9126","authenticated-orcid":false,"given":"Roberta","family":"Rizzo","sequence":"additional","affiliation":[{"name":"Department of Chemical and Pharmaceutical Science, University of Ferrara, 44121 Ferrara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1056\/NEJMoa2001017","article-title":"A Novel Coronavirus from Patients with Pneumonia in China, 2019","volume":"382","author":"Zhu","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1056\/NEJMoa2001316","article-title":"Early Transmission Dynamics in Wuhan, China, of Novel Coronavirus-Infected Pneumonia","volume":"382","author":"Li","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1038\/s41586-020-2012-7","article-title":"A pneumonia outbreak associated with a new coronavirus of probable bat origin","volume":"579","author":"Zhou","year":"2020","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1708","DOI":"10.1056\/NEJMoa2002032","article-title":"Clinical Characteristics of Coronavirus Disease 2019 in China","volume":"382","author":"Guan","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Qin, C., Zhou, L., Hu, Z., Zhang, S., Yang, S., Tao, Y., Xie, C., Ma, K., Shang, K., and Wang, W. (2020). Dysregulation of immune response in patients with COVID-19 in Wuhan, China. Clin. Infect. Dis.","DOI":"10.2139\/ssrn.3541136"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2202","DOI":"10.1172\/JCI137647","article-title":"SARS-CoV-2: A storm is raging","volume":"130","author":"Pedersen","year":"2020","journal-title":"J. Clin. Investig."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1038\/nbt.3860","article-title":"HLA-E-expressing pluripotent stem cells escape allogeneic responses and lysis by NK cells","volume":"35","author":"Gornalusse","year":"2017","journal-title":"Nat. Biotechnol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Tan, L., Wang, Q., Zhang, D., Ding, J., Huang, Q., Tang, Y.Q., Wang, Q., and Miao, H. (2020). Lymphopenia predicts disease severity of COVID-19: A descriptive and predictive study. Signal. Transduct. Target. Ther., 5.","DOI":"10.1038\/s41392-020-0148-4"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1038\/ni1582","article-title":"Functions of natural killer cells","volume":"9","author":"Vivier","year":"2008","journal-title":"Nat. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1146\/annurev-immunol-020711-075005","article-title":"Controlling natural killer cell responses: Integration of signals for activation and inhibition","volume":"31","author":"Long","year":"2013","journal-title":"Annu. Rev. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3603","DOI":"10.4049\/jimmunol.158.8.3603","article-title":"Natural killer cell cytolytic activity is inhibited by NKG2-A and activated by NKG2-C","volume":"158","author":"Houchins","year":"1997","journal-title":"J. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1038\/bmt.2008.380","article-title":"HLA-E upregulation on IFN-gamma-activated AML blasts impairs CD94\/NKG2A-dependent NK cytolysis after haplo-mismatched hematopoietic SCT","volume":"43","author":"Nguyen","year":"2009","journal-title":"Bone Marrow Transplant."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.4049\/jimmunol.171.3.1369","article-title":"Analysis of HLA-E peptide-binding specificity and contact residues in bound peptide required for recognition by CD94\/NKG2","volume":"171","author":"Miller","year":"2003","journal-title":"J. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1002\/eji.1830270517","article-title":"The human major histocompatibility complex class Ib molecule HLA-E binds signal sequence-derived peptides with primary anchor residues at positions 2 and 9","volume":"27","author":"Braud","year":"1997","journal-title":"Eur. J. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5199","DOI":"10.1073\/pnas.95.9.5199","article-title":"HLA-E is a major ligand for the natural killer inhibitory receptor CD94\/NKG2A","volume":"95","author":"Lee","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4741","DOI":"10.4049\/jimmunol.157.11.4741","article-title":"Human natural killer cell receptors involved in MHC class I recognition are disulfide-linked heterodimers of CD94 and NKG2 subunits","volume":"157","author":"Lazetic","year":"1996","journal-title":"J. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Marci, R., Gentili, V., Bortolotti, D., Lo Monte, G., Caselli, E., Bolzani, S., Rotola, A., Di Luca, D., and Rizzo, R. (2016). Presence of HHV-6A in Endometrial Epithelial Cells from Women with Primary Unexplained Infertility. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0158304"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Mirjacic Martinovic, K., Babovic, N., Dzodic, R., Jurisic, V., Matkovic, S., and Konjevic, G. (2015). Favorable in vitro effects of combined IL-12 and IL-18 treatment on NK cell cytotoxicity and CD25 receptor expression in metastatic melanoma patients. J. Transl. Med., 13.","DOI":"10.1186\/s12967-015-0479-z"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Nomura, S., Takahashi, H., Suzuki, J., Kuwahara, M., Yamashita, M., and Sawasaki, T. (2019). Pyrrothiogatain acts as an inhibitor of GATA family proteins and inhibits Th2 cell differentiation in vitro. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-53856-1"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Bortolotti, D., Gentili, V., Rotola, A., Cultrera, R., Marci, R., Di Luca, D., and Rizzo, R. (2019). HHV-6A infection of endometrial epithelial cells affects immune profile and trophoblast invasion. Am. J. Reprod. Immunol., 82.","DOI":"10.1111\/aji.13174"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Bortolotti, D., Gentili, V., Caselli, E., Sicolo, M., Soffritti, I., D\u2019Accolti, M., Barao, I., Rotola, A., Di Luca, D., and Rizzo, R. (2020). DNA Sensors\u2019 Signaling in NK Cells During HHV-6A, HHV-6B and HHV-7 Infection. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.00226"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2889","DOI":"10.4049\/jimmunol.171.6.2889","article-title":"Activating and inhibitory Ly49 receptors modulate NK cell chemotaxis to CXC chemokine ligand (CXCL) 10 and CXCL12","volume":"171","author":"Inngjerdingen","year":"2003","journal-title":"J. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.jneuroim.2012.07.004","article-title":"Altered natural killer cells\u2019 response to herpes virus infection in multiple sclerosis involves KIR2DL2 expression","volume":"251","author":"Rizzo","year":"2012","journal-title":"J. Neuroimmunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"F9","DOI":"10.1084\/jem.20012040","article-title":"Dendritic and natural killer cells cooperate in the control\/switch of innate immunity","volume":"195","author":"Zitvogel","year":"2002","journal-title":"J. Exp. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.3390\/v4091711","article-title":"Human leukocyte antigen (HLA) class I down-regulation by human immunodeficiency virus type 1 negative factor (HIV-1 Nef): What might we learn from natural sequence variants?","volume":"4","author":"Mwimanzi","year":"2012","journal-title":"Viruses"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Stevens, J., Joly, E., Trowsdale, J., and Butcher, G.W. (2001). Peptide binding characteristics of the non-classical class Ib MHC molecule HLA-E assessed by a recombinant random peptide approach. BMC Immunol., 2.","DOI":"10.1186\/1471-2172-2-5"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1183\/09031936.01.00040701","article-title":"Expression of GATA family of transcription factors in T-cells, monocytes and bronchial biopsies","volume":"18","author":"Caramori","year":"2001","journal-title":"Eur. Respir. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/S0092-8674(00)80240-8","article-title":"The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells","volume":"89","author":"Zheng","year":"1997","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3506","DOI":"10.4049\/jimmunol.1102635","article-title":"Upregulated protein arginine methyltransferase 1 by IL-4 increases eotaxin-1 expression in airway epithelial cells and participates in antigen-induced pulmonary inflammation in rats","volume":"188","author":"Sun","year":"2012","journal-title":"J. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1038\/s41423-020-0402-2","article-title":"Functional exhaustion of antiviral lymphocytes in COVID-19 patients","volume":"17","author":"Zheng","year":"2020","journal-title":"Cell Mol. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Antonioli, L., Fornai, M., Pellegrini, C., and Blandizzi, C. (2020). NKG2A and COVID-19: Another brick in the wall. Cell Mol. Immunol.","DOI":"10.1038\/s41423-020-0450-7"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Yaqinuddin, A., and Kashir, J. (2020). Innate immunity in COVID-19 patients mediated by NKG2A receptors, and potential treatment using Monalizumab, Cholroquine, and antiviral agents. Med. Hypotheses, 140.","DOI":"10.1016\/j.mehy.2020.109777"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Jewett, A. (2020). The Potential Effect of Novel Coronavirus SARS-CoV-2 on NK Cells; A Perspective on Potential Therapeutic Interventions. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01692"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Market, M.A.L., Martel, A.B., Bastin, D., Olanubi, O., Tennakoon, G., Boucher, D.M., Ng, J., Ardolino, M., and Auer, R.C. (2020). Flattening the COVID-19 Curve With Natural Killer Cell Based Immunotherapies. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01512"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.cell.2020.02.052","article-title":"SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor","volume":"181","author":"Hoffmann","year":"2020","journal-title":"Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/j.virol.2006.06.011","article-title":"Interferon-gamma and interleukin-4 downregulate expression of the SARS coronavirus receptor ACE2 in Vero E6 cells","volume":"353","author":"Osterhaus","year":"2006","journal-title":"Virology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.virusres.2007.03.015","article-title":"SARS coronavirus and innate immunity","volume":"133","author":"Frieman","year":"2008","journal-title":"Virus Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.immuni.2006.06.013","article-title":"Human NK cell education by inhibitory receptors for MHC class I","volume":"25","author":"Anfossi","year":"2006","journal-title":"Immunity"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Grote, D., Boualia, S.K., Souabni, A., Merkel, C., Chi, X., Costantini, F., Carroll, T., and Bouchard, M. (2008). Gata3 acts downstream of beta-catenin signaling to prevent ectopic metanephric kidney induction. PLoS Genet., 4.","DOI":"10.1371\/journal.pgen.1000316"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1111\/cea.12022","article-title":"Novel cooperation between CX3CL1 and CCL26 inducing NK cell chemotaxis via CX3CR1: A possible mechanism for NK cell infiltration of the allergic nasal tissue","volume":"43","author":"Doloriert","year":"2013","journal-title":"Clin. Exp. Allergy"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1528","DOI":"10.1182\/blood-2005-05-2073","article-title":"Engagement of CD83 ligand induces prolonged expansion of CD8+ T cells and preferential enrichment for antigen specificity","volume":"107","author":"Hirano","year":"2006","journal-title":"Blood"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.bbadis.2010.12.013","article-title":"ER stress impairs MHC Class I surface expression and increases susceptibility of thyroid cells to NK-mediated cytotoxicity","volume":"1812","author":"Ulianich","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"van Hall, T., Andre, P., Horowitz, A., Ruan, D.F., Borst, L., Zerbib, R., Narni-Mancinelli, E., van der Burg, S.H., and Vivier, E. (2019). Monalizumab: Inhibiting the novel immune checkpoint NKG2A. J. Immunother. Cancer, 7.","DOI":"10.1186\/s40425-019-0761-3"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Masselli, E.V.M., Carubbi, C., Pozzi, G., Presta, V., Mirandola, P., and Vitale, M. (2020). NK cells: A double edge sword against SARS-CoV-2. Adv. Biol. Regul., 77.","DOI":"10.1016\/j.jbior.2020.100737"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/1975\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:07:18Z","timestamp":1760177238000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/1975"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,26]]},"references-count":44,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["cells9091975"],"URL":"https:\/\/doi.org\/10.3390\/cells9091975","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-31860\/v1","asserted-by":"object"},{"id-type":"doi","id":"10.21203\/rs.3.pex-959\/v1","asserted-by":"object"}]},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,26]]}}}