{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T06:17:38Z","timestamp":1790835458203,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,5,23]],"date-time":"2022-05-23T00:00:00Z","timestamp":1653264000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In glioblastoma, non-classical human leucocyte antigen E (HLA-E) and HLA-G are frequently overexpressed. HLA-E loaded with peptides derived from HLA class I and from HLA-G contributes to inhibition of natural killer (NK) cells with expression of the inhibitory receptor CD94\/NKG2A. We investigated whether NK cells expressing the activating CD94\/NKG2C receptor counterpart were able to exert anti-glioma effects. NKG2C+ subsets were preferentially expanded by a feeder cell line engineered to express an artificial disulfide-stabilized trimeric HLA-E ligand (HLA-E*spG). NK cells expanded by a feeder cell line, which facilitates outgrowth of conventional NKG2A+, and fresh NK cells, were included for comparison. Expansion via the HLA-E*spG feeder cells selectively increased the fraction of NKG2C+ NK cells, which displayed a higher frequency of KIR2DL2\/L3\/S2 and CD16 when compared to expanded NKG2A+ NK cells. NKG2C+ NK cells exhibited increased cytotoxicity against K562 and KIR:HLA-matched and -mismatched primary glioblastoma multiforme (GBM) cells when compared to NKG2A+ NK cells and corresponding fresh NK cells. Cytotoxic responses of NKG2C+ NK cells were even more pronounced when utilizing target cells engineered with HLA-E*spG. These findings support the notion that NKG2C+ NK cells have potential therapeutic value for treating gliomas.<\/jats:p>","DOI":"10.3390\/ijms23105857","type":"journal-article","created":{"date-parts":[[2022,5,23]],"date-time":"2022-05-23T22:57:22Z","timestamp":1653346642000},"page":"5857","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["NKG2C+ NK Cells for Immunotherapy of Glioblastoma Multiforme"],"prefix":"10.3390","volume":"23","author":[{"given":"Shafiq","family":"Murad","sequence":"first","affiliation":[{"name":"Department of Neurosurgery, Section Experimental Neurosurgery\/Tumor Immunology, University Hospital Carl Gustav Carus, TU Dresden, Fetscherstr. 74, 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5726-386X","authenticated-orcid":false,"given":"Susanne","family":"Michen","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Section Experimental Neurosurgery\/Tumor Immunology, University Hospital Carl Gustav Carus, TU Dresden, Fetscherstr. 74, 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Becker","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Section Experimental Neurosurgery\/Tumor Immunology, University Hospital Carl Gustav Carus, TU Dresden, Fetscherstr. 74, 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monika","family":"F\u00fcssel","sequence":"additional","affiliation":[{"name":"DKMS Life Science Lab GmbH, St. Petersburger Str. 2, 01069 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriele","family":"Schackert","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Section Experimental Neurosurgery\/Tumor Immunology, University Hospital Carl Gustav Carus, TU Dresden, Fetscherstr. 74, 01307 Dresden, Germany"},{"name":"German Cancer Consortium (DKTK), Partner Site Dresden and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"National Center for Tumor Diseases (NCT\/UCC), 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Torsten","family":"Tonn","sequence":"additional","affiliation":[{"name":"Department of Transfusion Medicine, Medical Faculty Carl Gustav Carus, Technische Universit\u00e4t Dresden, Fetscherstr. 74, 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frank","family":"Momburg","sequence":"additional","affiliation":[{"name":"Antigen Presentation and T\/NK Cell Activation Group (D121), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany"},{"name":"Clinical Cooperation Unit \u201cApplied Tumor Immunity\u201d (D120), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5084-1180","authenticated-orcid":false,"given":"Achim","family":"Temme","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, Section Experimental Neurosurgery\/Tumor Immunology, University Hospital Carl Gustav Carus, TU Dresden, Fetscherstr. 74, 01307 Dresden, Germany"},{"name":"German Cancer Consortium (DKTK), Partner Site Dresden and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"National Center for Tumor Diseases (NCT\/UCC), 01307 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1007\/s00401-016-1545-1","article-title":"The 2016 World Health Organization Classification of Tumors of the Central Nervous System: A summary","volume":"131","author":"Louis","year":"2016","journal-title":"Acta Neuropathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/S1470-2045(09)70025-7","article-title":"Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial","volume":"10","author":"Stupp","year":"2009","journal-title":"Lancet Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1200\/JCO.2016.70.7562","article-title":"Radiation Therapy for Glioblastoma: American Society of Clinical Oncology Clinical Practice Guideline Endorsement of the American Society for Radiation Oncology Guideline","volume":"35","author":"Sulman","year":"2017","journal-title":"J. Clin. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1373","DOI":"10.1016\/S1470-2045(17)30517-X","article-title":"Rindopepimut with temozolomide for patients with newly diagnosed, EGFRvIII-expressing glioblastoma (ACT IV): A randomised, double-blind, international phase 3 trial","volume":"18","author":"Weller","year":"2017","journal-title":"Lancet Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1093\/neuonc\/nox208","article-title":"Nivolumab with or without ipilimumab in patients with recurrent glioblastoma: Results from exploratory phase I cohorts of CheckMate 143","volume":"20","author":"Omuro","year":"2018","journal-title":"Neuro Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1001\/jamaoncol.2020.1024","article-title":"Effect of Nivolumab vs Bevacizumab in Patients with Recurrent Glioblastoma the CheckMate 143 Phase 3 Randomized Clinical Trial","volume":"6","author":"Reardon","year":"2020","journal-title":"JAMA Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1038\/nature12477","article-title":"Signatures of mutational processes in human cancer","volume":"500","author":"Alexandrov","year":"2013","journal-title":"Nature"},{"key":"ref_8","first-page":"35","article-title":"Prospects of immune checkpoint blockade and vaccine-based immunotherapy for glioblastoma","volume":"6","author":"Tietze","year":"2021","journal-title":"Innov. Surg. Sci."},{"key":"ref_9","first-page":"918471","article-title":"Tumor evasion from T cell surveillance","volume":"2011","author":"Topfer","year":"2011","journal-title":"J. Biomed. Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"eaaa0984","DOI":"10.1126\/scitranslmed.aaa0984","article-title":"A single dose of peripherally infused EGFRvIII-directed CAR T cells mediates antigen loss and induces adaptive resistance in patients with recurrent glioblastoma","volume":"9","author":"Nasrallah","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2561","DOI":"10.1056\/NEJMoa1610497","article-title":"Regression of Glioblastoma after Chimeric Antigen Receptor T-Cell Therapy","volume":"375","author":"Brown","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/nrc.2015.5","article-title":"NK cells and cancer: You can teach innate cells new tricks","volume":"16","author":"Morvan","year":"2015","journal-title":"Nat. Rev. Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1615\/CritRevImmunol.2017019376","article-title":"Genetically Engineered Natural Killer Cells as a Means for Adoptive Tumor Immunotherapy","volume":"36","author":"Michen","year":"2016","journal-title":"Crit. Rev. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.smim.2014.02.007","article-title":"NK cell self tolerance, responsiveness and missing self recognition","volume":"26","author":"Shifrin","year":"2014","journal-title":"Semin. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1084\/jem.20072525","article-title":"CD94-NKG2A recognition of human leukocyte antigen (HLA)-E bound to an HLA class I leader sequence","volume":"205","author":"Petrie","year":"2008","journal-title":"J. Exp. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1016\/S1074-7613(00)80574-9","article-title":"Association of DAP12 with activating CD94\/NKG2C NK cell receptors","volume":"8","author":"Lanier","year":"1998","journal-title":"Immunity"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1084\/jem.20042558","article-title":"Natural selection drives recurrent formation of activating killer cell immunoglobulin-like receptor and Ly49 from inhibitory homologues","volume":"201","author":"Parham","year":"2005","journal-title":"J. Exp. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"13224","DOI":"10.1073\/pnas.0503594102","article-title":"Recognition of peptide-MHC class I complexes by activating killer immunoglobulin-like receptors","volume":"102","author":"Stewart","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1093\/jnen\/64.6.523","article-title":"HLA-E protects glioma cells from NKG2D-mediated immune responses in vitro: Implications for immune escape in vivo","volume":"64","author":"Wischhusen","year":"2005","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jneuroim.2012.05.010","article-title":"HLA-E contributes to an immune-inhibitory phenotype of glioblastoma stem-like cells","volume":"250","author":"Wolpert","year":"2012","journal-title":"J. Neuroimmunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1016\/j.ajpath.2012.10.021","article-title":"Human leukocyte antigen-G is frequently expressed in glioblastoma and may be induced in vitro by combined 5-aza-2\u2019-deoxycytidine and interferon-gamma treatments: Results from a multicentric study","volume":"182","author":"Wastowski","year":"2013","journal-title":"Am. J. Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1967","DOI":"10.1016\/j.celrep.2018.07.069","article-title":"Distinct HLA-E Peptide Complexes Modify Antibody-Driven Effector Functions of Adaptive NK Cells","volume":"24","author":"Rolle","year":"2018","journal-title":"Cell Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4250","DOI":"10.1093\/emboj\/18.15.4250","article-title":"Kinetics and peptide dependency of the binding of the inhibitory NK receptor CD94\/NKG2-A and the activating receptor CD94\/NKG2-C to HLA-E","volume":"18","author":"Reyburn","year":"1999","journal-title":"EMBO J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/35642","article-title":"Immunoreceptor DAP12 bearing a tyrosine-based activation motif is involved in activating NK cells","volume":"391","author":"Lanier","year":"1998","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3201","DOI":"10.4049\/jimmunol.1400330","article-title":"DAP12-Based Activating Chimeric Antigen Receptor for NK Cell Tumor Immunotherapy","volume":"194","author":"Topfer","year":"2015","journal-title":"J. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1097\/CJI.0000000000000082","article-title":"Engineering NK Cells Modified with an EGFRvIII-specific Chimeric Antigen Receptor to Overexpress CXCR4 Improves Immunotherapy of CXCL12\/SDF-1alpha-secreting Glioblastoma","volume":"38","author":"Muller","year":"2015","journal-title":"J. Immunother."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3664","DOI":"10.1182\/blood-2004-05-2058","article-title":"Imprint of human cytomegalovirus infection on the NK cell receptor repertoire","volume":"104","author":"Guma","year":"2004","journal-title":"Blood"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/s41590-018-0082-6","article-title":"Peptide-specific recognition of human cytomegalovirus strains controls adaptive natural killer cells","volume":"19","author":"Hammer","year":"2018","journal-title":"Nat. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3624","DOI":"10.1182\/blood-2005-09-3682","article-title":"Expansion of CD94\/NKG2C+ NK cells in response to human cytomegalovirus-infected fibroblasts","volume":"107","author":"Guma","year":"2006","journal-title":"Blood"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100870","DOI":"10.1016\/j.mam.2020.100870","article-title":"NK cells and ILCs in tumor immunotherapy","volume":"80","author":"Sivori","year":"2021","journal-title":"Mol. Asp. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.jcyt.2020.02.004","article-title":"Artificial feeder cells expressing ligands for killer cell immunoglobulin-like receptors and CD94\/NKG2A for expansion of functional primary natural killer cells with tolerance to self","volume":"22","author":"Michen","year":"2020","journal-title":"Cytotherapy"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1016\/j.humimm.2015.09.003","article-title":"HLA-E regulates NKG2C+natural killer cell function through presentation of a restricted peptide repertoire","volume":"76","author":"Lauterbach","year":"2015","journal-title":"Hum. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2557","DOI":"10.1084\/jem.20091010","article-title":"KIR2DS4 is a product of gene conversion with KIR3DL2 that introduced specificity for HLA-A*11 while diminishing avidity for HLA-C","volume":"206","author":"Graef","year":"2009","journal-title":"J. Exp. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2678","DOI":"10.1182\/blood-2012-10-459545","article-title":"NK cell responses to cytomegalovirus infection lead to stable imprints in the human KIR repertoire and involve activating KIRs","volume":"121","author":"Beziat","year":"2013","journal-title":"Blood"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.immuni.2015.02.008","article-title":"Cytomegalovirus Infection Drives Adaptive Epigenetic Diversification of NK Cells with Altered Signaling and Effector Function","volume":"42","author":"Schlums","year":"2015","journal-title":"Immunity"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1869","DOI":"10.3389\/fimmu.2018.01869","article-title":"Natural Killer Cells: Development, Maturation, and Clinical Utilization","volume":"9","author":"Abel","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_37","first-page":"331","article-title":"Natural-Killer-Cell Immunodeficiency in Hiv Disease Is Manifest by Profoundly Decreased Numbers of Cd16(+)Cd56(+) Cells and Expansion of a Population of Cd16(Dim)Cd56(-) Cells with Low Lytic Activity","volume":"10","author":"Hu","year":"1995","journal-title":"J. Acq. Immun. Def. Synd."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6612","DOI":"10.4049\/jimmunol.0901437","article-title":"Expansion of Functionally Skewed CD56-Negative NK Cells in Chronic Hepatitis C Virus Infection: Correlation with Outcome of Pegylated IFN-alpha and Ribavirin Treatment","volume":"183","author":"Gonzalez","year":"2009","journal-title":"J. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"724","DOI":"10.18632\/aging.101774","article-title":"CD56-negative NK cells with impaired effector function expand in CMV and EBV co-infected healthy donors with age","volume":"11","author":"Grahlert","year":"2019","journal-title":"Aging"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3770","DOI":"10.1172\/JCI125916","article-title":"Chronic stimulation drives human NK cell dysfunction and epigenetic reprograming","volume":"129","author":"Merino","year":"2019","journal-title":"J. Clin. Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1158\/2326-6066.CIR-17-0207","article-title":"Intrinsic Functional Potential of NK-Cell Subsets Constrains Retargeting Driven by Chimeric Antigen Receptors","volume":"6","author":"Oei","year":"2018","journal-title":"Cancer Immunol. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4951","DOI":"10.4049\/jimmunol.160.10.4951","article-title":"HLA-E surface expression depends on binding of TAP-dependent peptides derived from certain HLA class I signal sequences","volume":"160","author":"Lee","year":"1998","journal-title":"J. Immunol."},{"key":"ref_43","first-page":"8996","article-title":"MICA\/NKG2D-mediated immunogene therapy of experimental gliomas","volume":"63","author":"Friese","year":"2003","journal-title":"Cancer Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5326","DOI":"10.1158\/0008-5472.CAN-16-1162","article-title":"Identification of a Natural Killer Cell Receptor Allele That Prolongs Survival of Cytomegalovirus-Positive Glioblastoma Patients","volume":"76","author":"Gras","year":"2016","journal-title":"Cancer Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1074","DOI":"10.1093\/neuonc\/nor098","article-title":"RNA interference targeting survivin exerts antitumoral effects in vitro and in established glioma xenografts in vivo","volume":"13","author":"Hendruschk","year":"2011","journal-title":"Neuro Oncol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/10\/5857\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:17:20Z","timestamp":1760138240000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/10\/5857"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,23]]},"references-count":45,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,5]]}},"alternative-id":["ijms23105857"],"URL":"https:\/\/doi.org\/10.3390\/ijms23105857","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,23]]}}}