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Freshly isolated NK cells exhibit low cytolytic activity. Therefore, efficient activation of NK cells is essential to enhance their anti-cancer properties. The aim of this study was to compare ex vivo NK cells activation with commonly used cytokines, including IL-2, IL-12, IL-15, IL-18, or STING agonist, and to determine the most effective activation method. Our results indicate higher viability and expression of activating receptors and granzyme B in IL-15-treated NK cells. The addition of IL-12 to IL-15 augmented the production of IFN-\u03b3 and CCL3 in NK cells. However, incubation of NK cells with a cocktail of IL-12, IL-15, and IL-18 cytokines generated massive production of pro-inflammatory cytokines and a significant increase in activation marker CD69. We have also observed reduced viability of NK cells after incubation with STING agonist. NK cells activated with IL-12\/IL-15, and IL-12\/IL-15\/IL-18 co-cultured with MHC\n                    <jats:sup>\u2212<\/jats:sup>\n                    tumor cells, and in each cytokine combination with MHC\n                    <jats:sup>+<\/jats:sup>\n                    tumor cells caused proliferation inhibition and lysis of tumor cells. Activation of NK cells, particularly with the combination of IL-12\/IL-15\/IL-18 cytokines, shows potential clinical relevance in adoptive cell therapy.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-42816-1","type":"journal-article","created":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T17:16:04Z","timestamp":1773681364000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Characteristics of NK cells activation following IL-12, IL-15, IL-18 cytokines stimulation for preclinical purposes"],"prefix":"10.1038","volume":"16","author":[{"given":"Justyna","family":"Czapla","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alina","family":"Drzyzga","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryszard","family":"Smolarczyk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,3,16]]},"reference":[{"key":"42816_CR1","doi-asserted-by":"publisher","first-page":"1319","DOI":"10.1002\/JLB.MR0718-269R","volume":"105","author":"I Prager","year":"2019","unstructured":"Prager, I. & Watzl, C. 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The animal experiment was approved by the Local Ethical Committee for Experiments on Animals at the Medical University of Silesia in Katowice (decision No. 6\/2024).","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics declarations"}}],"article-number":"13665"}}