{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T08:28:21Z","timestamp":1787300901617,"version":"build-2736575974"},"reference-count":16,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T00:00:00Z","timestamp":1780531200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100002424","name":"FUJIFILM Corporation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002424","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005991","name":"Pharmaceutical and Medical Device Regulatory Science Society of Japan","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005991","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Pyrogen testing is essential for ensuring pharmaceutical product safety; however, current methods have limitations. The Bacterial Endotoxins Test cannot detect non-endotoxin pyrogens, whereas the Rabbit pyrogen test was phased out for animal welfare reasons. Monocyte activation tests (MAT), using human whole blood samples or peripheral blood mononuclear cells, are effective but suffer from donor variability, biosafety concerns and instability. We aimed to develop a sensitive, rapid and simple alternative pyrogen test by devising a luciferase-based MAT using the Mono-Mac-6 (MM6) cell line. MM6 cells showed higher cytokine responses to endotoxin than THP-1 cells and exhibited comparable or greater sensitivity than human peripheral blood. Introducing an NF-\u03baB-driven NanoLuc\u00ae reporter enabled endotoxin detection within 3\u00a0h; however, rapid degradation of intracellular luciferase was noted. To overcome this degradation issue, we generated MM6 cells expressing secreted NanoLuc (secNluc), allowing for stable detection between 3 and 12\u00a0h after stimulation. The MM6\/NF\u03baB-secNluc cells helped detect various pyrogens, including Pam3CSK4, peptidoglycan, polyinosinic-polycytidylic acid sodium salt, lipopolysaccharide, flagellin, macrophage-activating lipopeptide-2 and resiquimod. Adding recombinant human lipopolysaccharide-binding protein enabled establishing a serum-free MAT system, eliminating serum-derived endotoxin contamination. This reporter-based MAT provides a robust platform for rapid endotoxin and NEP detection, offering an improved alternative to conventional MATs.<\/jats:p>","DOI":"10.1093\/jb\/mvag038","type":"journal-article","created":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T11:33:15Z","timestamp":1780399995000},"page":"207-215","source":"Crossref","is-referenced-by-count":0,"title":["Development and validation of a novel monocyte activation test with secreted luciferase-expressing cells"],"prefix":"10.1093","volume":"180","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7360-6642","authenticated-orcid":false,"given":"Teruaki","family":"Oku","sequence":"first","affiliation":[{"name":"Department of Microbiology, Hoshi University School of Pharmacy and Pharmaceutical Sciences , 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusuke","family":"Ando","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hoshi University School of Pharmacy and Pharmaceutical Sciences , 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501 ,","place":["Japan"]},{"name":"Laboratory of Clinical Pathology, Faculty of Pharmacy and Pharmaceutical Sciences School of Pharmacy, Josai University , 1-1 Keyaki-dai, Sakado, Saitama 350-0295 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohisa","family":"Nanao","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hoshi University School of Pharmacy and Pharmaceutical Sciences , 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501 ,","place":["Japan"]},{"name":"Bio Science & Engineering Laboratories, FUJIFILM Corporation , 6-1 Takata-cho, Amagasaki, Hyogo 661-0963 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yutaka","family":"Kikuchi","sequence":"additional","affiliation":[{"name":"Department of Nutrition, Faculty of Healthcare Sciences, Chiba Prefectural University of Health Sciences , 2-10-1 Wakaba, Mihama-ku, Chiba, Chiba 261-0014 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukiko","family":"Hara-Kudo","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hoshi University School of Pharmacy and Pharmaceutical Sciences , 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,6,4]]},"reference":[{"key":"2026082104211678400_ref1","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/978-3-319-92336-9_3","volume-title":"Clinical Manual of Fever in Children","author":"El-Radhi","year":"2018"},{"key":"2026082104211678400_ref2","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1128\/CMR.8.2.268","article-title":"Endotoxemia: methods of detection and clinical correlates","volume":"8","author":"Hurley","year":"1995","journal-title":"Clin. 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