{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T12:29:56Z","timestamp":1790512196376,"version":"4.1.0"},"reference-count":44,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,12,27]],"date-time":"2021-12-27T00:00:00Z","timestamp":1640563200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006342","name":"Vaincre la Mucoviscidose","doi-asserted-by":"publisher","award":["RF20190502487"],"award-info":[{"award-number":["RF20190502487"]}],"id":[{"id":"10.13039\/501100006342","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Insttitut de Recherche en sant\u00e9 respiratoire des Pays de la Loire","award":["LSC 10280"],"award-info":[{"award-number":["LSC 10280"]}]},{"name":"Association Gr\u00e9gory Lemarchal","award":["RF20190502487"],"award-info":[{"award-number":["RF20190502487"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>HCMV drives complex and multiple cellular immune responses, which causes a persistent immune imprint in hosts. This study aimed to achieve both a quantitative determination of the frequency for various anti-HCMV immune cell subsets, including CD8 T, \u03b3\u03b4T, NK cells, and a qualitative analysis of their phenotype. To map the various anti-HCMV cellular responses, we used a combination of three HLApeptide tetramer complexes (HLA-EVMAPRTLIL, HLA-EVMAPRSLLL, and HLA-A2NLVPMVATV) and antibodies for 18 surface markers (CD3, CD4, CD8, CD16, CD19, CD45RA, CD56, CD57, CD158, NKG2A, NKG2C, CCR7, TCR\u03b3\u03b4, TCR\u03b3\u03b42, CX3CR1, KLRG1, 2B4, and PD-1) in a 20-color spectral flow cytometry analysis. This immunostaining protocol was applied to PBMCs isolated from HCMV\u2212 and HCMV+ individuals. Our workflow allows the efficient determination of events featuring HCMV infection such as CD4\/CD8 ratio, CD8 inflation and differentiation, HCMV peptide-specific HLA-EUL40 and HLA-A2pp65CD8 T cells, and expansion of \u03b3\u03b4T and NK subsets including \u03b42\u2212\u03b3T and memory-like NKG2C+CD57+ NK cells. Each subset can be further characterized by the expression of 2B4, PD-1, KLRG1, CD45RA, CCR7, CD158, and NKG2A to achieve a fine-tuned mapping of HCMV immune responses. This assay should be useful for the analysis and monitoring of T-and NK cell responses to HCMV infection or vaccines.<\/jats:p>","DOI":"10.3390\/ijms23010263","type":"journal-article","created":{"date-parts":[[2021,12,28]],"date-time":"2021-12-28T01:18:15Z","timestamp":1640654295000},"page":"263","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Mapping and Characterization of HCMV-Specific Unconventional HLA-E-Restricted CD8 T Cell Populations and Associated NK and T Cell Responses Using HLA\/Peptide Tetramers and Spectral Flow Cytometry"],"prefix":"10.3390","volume":"23","author":[{"given":"Am\u00e9lie","family":"Rousseli\u00e8re","sequence":"first","affiliation":[{"name":"INSERM, Center for Research in Transplantation and Translational Immunology, Nantes Universit\u00e9, UMR 1064, CHU Nantes, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laurence","family":"Delbos","sequence":"additional","affiliation":[{"name":"INSERM, Center for Research in Transplantation and Translational Immunology, Nantes Universit\u00e9, UMR 1064, CHU Nantes, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5175-0790","authenticated-orcid":false,"given":"C\u00e9line","family":"Bressollette","sequence":"additional","affiliation":[{"name":"INSERM, Center for Research in Transplantation and Translational Immunology, Nantes Universit\u00e9, UMR 1064, CHU Nantes, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ma\u00eflys","family":"Berthaume","sequence":"additional","affiliation":[{"name":"INSERM, Center for Research in Transplantation and Translational Immunology, Nantes Universit\u00e9, UMR 1064, CHU Nantes, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5870-4615","authenticated-orcid":false,"given":"B\u00e9atrice","family":"Charreau","sequence":"additional","affiliation":[{"name":"INSERM, Center for Research in Transplantation and Translational Immunology, Nantes Universit\u00e9, UMR 1064, CHU Nantes, F-44000 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1673","DOI":"10.1172\/JCI45449","article-title":"Cytomegalovirus: Pathogen, paradigm, and puzzle","volume":"121","author":"Boeckh","year":"2011","journal-title":"J. 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