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Activated iNKT cells are known to exert anti-tumor effects through the activation of other immune cells and have attracted attention as promising T cells for eliciting anti-tumor immunity. However, securing a sufficient number of iNKT cells is an obstacle to treatment because iNKT cells are a very small cell population, less than 0.1% of the peripheral blood lymphocytes. Although previous studies have demonstrated redifferentiation of a large number of CD4\n                    <jats:sup>\u2013<\/jats:sup>\n                    CD8\n                    <jats:sup>\u2013<\/jats:sup>\n                    double-negative iNKT cells from induced pluripotent stem (iPS) cells in two-dimensional monolayer cultures, CD4\n                    <jats:sup>+<\/jats:sup>\n                    single-positive (CD4SP) iNKT cells could not be induced. Here we show CD4SP iNKT cells can be obtained by three-dimensional (3D) organoid culture (3D-CD4+ iNKT cell). We additionally describe 3D-CD4+ iNKT cells show antigen-specific helper functions, as they proliferate, produce interferon-\u03b3\/interleukin-4 (IFN-\u03b3\/IL-4), and induce dendritic cell maturation in response to \u03b1-galactosylceramide. Furthermore, they reverse the inhibition of T cell proliferation induced by immunosuppressive macrophages in an antigen-specific manner. Collectively, 3D-CD4+ iNKT cells may become an adjuvant T-cell source to enhance current T-cell immunotherapy against solid tumors.\n                  <\/jats:p>","DOI":"10.1038\/s42003-025-09462-1","type":"journal-article","created":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T12:35:14Z","timestamp":1767962114000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Artificial thymic organoid culture generates functional iPSC-derived CD4+ invariant natural killer T cells"],"prefix":"10.1038","volume":"9","author":[{"given":"Sara","family":"Shiina","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6705-5626","authenticated-orcid":false,"given":"Tatsuki","family":"Ueda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-9228-2352","authenticated-orcid":false,"given":"Shoichi","family":"Iriguchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasushi","family":"Uemura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2291-4586","authenticated-orcid":false,"given":"Shin","family":"Kaneko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,1,9]]},"reference":[{"key":"9462_CR1","doi-asserted-by":"publisher","first-page":"715","DOI":"10.1038\/nri1936","volume":"6","author":"L Zitvogel","year":"2006","unstructured":"Zitvogel, L., Tesniere, A. & Kroemer, G. 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