{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T16:04:44Z","timestamp":1779465884168,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,19]]},"group-title":"Bioengineering","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T00:00:00Z","timestamp":1779148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>The limiting step in immune-active drug development is increasingly not candidate generation but testing whether a candidate therapy is effective in a system that preserves tissue architecture, vascular exposure, multicellular interaction, and repeated pharmacodynamic sampling without patient exposure. We developed an acellular normothermic machine-perfusion platform for intact porcine spleen designed as a translational immune-organ assay. Across independent acellular perfusions, the circuit maintained physiologic parameters, preserved red- and white-pulp histology, and yielded viable effluent cells suitable for serial flow cytometry and multiomics. High-dose methylprednisolone was used as a clinically familiar perturbation to determine whether the platform could resolve steroid immunosuppression at mechanistically distinct levels. Effluent RNA-seq identified canonical glucocorticoid-responsive transcriptional programs, including DUSP1, FKBP5, PER1, DDIT4, SGK1, KLF9, ANXA1, NF-\u03baB feedback regulators, and JAK\/STAT suppressor pathways. SOCS3 was a prominent early signal in the perfusion transcriptome and was validated orthogonally at the protein level in prednisone-treated, CD3\/CD28-activated primary murine splenocytes, strengthening its role as a candidate pharmacodynamic marker. In parallel, data independent acquisition (DIA) proteomics of effluent cell pellets nominated a non-transcriptional protein-level response: a Sus scrofa LGALS13-annotated, CLC\/Galectin-10-like galectin detected despite absence of the corresponding effluent-cell transcript. Because this porcine LGALS13-annotated protein group is treated here as an orthologous CLC\/Galectin-10-like signal rather than as canonical human placental Galectin-13\/PP13, we tested recombinant human Galectin-10 in vitro. Human Galectin-10 induced marked apoptosis of CD3\/CD28-stimulated Jurkat cells, prioritizing this axis for future mechanistic testing without proving causality in the perfused spleen. These data establish acellular spleen perfusion as a serial multiomic platform for translational immunopharmacology and motivate deployment with otherwise-discarded human donor spleens.<\/jats:p>\n                <jats:sec>\n                  <jats:title>One sentence summary<\/jats:title>\n                  <jats:p>An acellular intact-spleen perfusion platform enables serial cellular, transcriptomic, proteomic, and functional pharmacodynamic sampling that identifies steroid-responsive transcriptional programs, validates SOCS3 protein induction, and nominates a CLC\/Galectin-10-like non-transcriptional immunosuppressive axis for translation to discarded human donor spleens.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.16.725632","type":"posted-content","created":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T19:40:16Z","timestamp":1779219616000},"source":"Crossref","is-referenced-by-count":0,"title":["Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression"],"prefix":"10.64898","author":[{"given":"Nicholas A.","family":"Callais","sequence":"first","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shawn","family":"Welch","sequence":"additional","affiliation":[{"name":"Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Randall R.","family":"Rainwater","sequence":"additional","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Schaller","sequence":"additional","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuoqiu","family":"Deng","sequence":"additional","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Azevedo-Pouly","sequence":"additional","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of America"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie Schluterman","family":"Burdine","sequence":"additional","affiliation":[{"name":"Division of Surgical Research, Department of Surgery, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas, United States of America"},{"name":"Arkansas Children\u2019s Research Institute, 13 Children\u2019s Way, Little Rock, Arkansas, United States of 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