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Imbalance in their distribution could occur in pathological conditions, including solid or haematological cancers, autoimmune disorders and metabolic diseases; therefore we aimed to define age\u2010matched reference values of the NKT\u2010like population. NKT\u2010like cell count and their expression of CD4 and CD8 surface antigens were evaluated in 2208 subjects by flow cytometry, with a lyse\u2010no\u2010wash protocol in a CE\u2010IVD workup. Outpatients without alteration of any laboratory test and adult healthy volunteers were used to determine NKT\u2010like age\u2010matched reference values; a total of 490 subjects divided into six age ranges were finally enrolled. We defined reference values for NKT\u2010like cells in adult and in 5 age\u2010matched paediatric control groups. NKT\u2010like cell number increases with age both in percentage and absolute count. While NKT\u2010like\/CD8+ cells increased with age, NKT\u2010like\/CD4+ cells showed an opposite trend. Similarly, double\u2010positive cells (NKT\u2010like\/CD4+ CD8+) gradually decreased with age, while double\u2010negative cells (NKT\u2010like\/CD4\u2010CD8\u2010) increased up to 16\u2009years and then decreased in adults. Ultimately, we evaluate adult and paediatric patients with NKT\u2010like cell count outside normal ranges derived here to highlight the principal associated conditions that may be involved. Our paper is a first step toward the definition of robust reference values for NKT\u2010like cell subpopulations under the CE\u2010IVD conditions. Providing NKT\u2010like cell number and frequency in the diagnostic report appears helpful to better delineate the immune profile of patients.<\/jats:p>","DOI":"10.1111\/sji.70062","type":"journal-article","created":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T06:32:21Z","timestamp":1761546741000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Age\u2010Matched Reference Values for Circulating Natural Killer T (\n                    <scp>NKT<\/scp>\n                    )\u2010Like Cells"],"prefix":"10.1111","volume":"102","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1933-6415","authenticated-orcid":false,"given":"Elena","family":"Trombetta","sequence":"first","affiliation":[{"name":"Flow Cytometry Laboratory, Clinical Pathology, Fondazione IRCCS ca' Granda Ospedale Maggiore Policlinico  Milan 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