{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T22:23:00Z","timestamp":1792621380957,"version":"4.5.0"},"reference-count":28,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T00:00:00Z","timestamp":1769385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>\n                      Chimeric antigen receptor (CAR) T-cell therapy has demonstrated remarkable efficacy in hematological malignancies. However, it can also cause severe systemic toxicity, known as cytokine release syndrome (CRS). Therefore, the potential of CAR-T cells to cause toxicity\n                      <jats:italic>in vivo<\/jats:italic>\n                      should be evaluated in preclinical models prior to first-in-human trials. Although murine models exist for this purpose, they are typically complex xenograft systems available only to a limited number of laboratories. Therefore, development of an\n                      <jats:italic>in vitro<\/jats:italic>\n                      assay to assess CRS elicited by CAR-T cells is warranted.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>CAR-T cells, macrophages, or immature dendritic cells (iDCs), along with tumor target cells, were co-cultured under different conditions. The release of CRS-related cytokines, IFN-\u03b3 and IL-6, was measured to simulate cytokine release during CAR-T-induced CRS. Additionally, the cellular source of the key CRS cytokine IL-6 was investigated.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      A co-culture system containing only CAR-T cells and tumor cells failed to recapitulate the key feature of CRS, specifically a significant elevation of IL-6. However, when CAR-T cells were co-cultured with antigen-presenting cells (macrophages or iDCs) and tumor cells, the core CRS cytokine IL-6 was significantly elevated in an\n                      <jats:italic>in vitro<\/jats:italic>\n                      cell culture model, indicating that this system effectively mimics cytokine release during CAR-T-induced CRS. Furthermore, macrophages and iDCs are the primary cellular sources of IL-6 during CRS, with macrophages playing a central role in the development of CRS. Additionally, a co-culture system involving CAR-T cells, tumor cells, and macrophages under these conditions can indicate the occurrence of clinically severe-grade CRS.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Macrophages and iDCs play a critical role in the development of CAR-T therapy-induced CRS. The triple-cell co-culture system, comprising CAR-T cells, macrophages or iDCs, and tumor cells, provides a viable\n                      <jats:italic>in vitro<\/jats:italic>\n                      model for assessing CAR-T cell-induced CRS.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1721114","type":"journal-article","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T07:20:06Z","timestamp":1769412006000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["An in vitro co-culture model with CAR-T cells, antigen-presenting cells, and tumor cells to evaluate CAR-T cell-induced cytokine release syndrome"],"prefix":"10.3389","volume":"16","author":[{"given":"Yuke","family":"Ren","sequence":"first","affiliation":[{"name":"National Institutes for Food and Drug Control, Chinese Academy of Medical Sciences and Peking Union Medical College","place":["Beijing, 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