{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T13:45:18Z","timestamp":1791639918576,"version":"4.3.4"},"reference-count":48,"publisher":"BMJ","issue":"7","license":[{"start":{"date-parts":[[2025,7,31]],"date-time":"2025-07-31T00:00:00Z","timestamp":1753920000000},"content-version":"unspecified","delay-in-days":30,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22494694"],"award-info":[{"award-number":["22494694"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82404500"],"award-info":[{"award-number":["82404500"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["W2412118"],"award-info":[{"award-number":["W2412118"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Science & Technology Commission of Shanghai Municipality","award":["23ZR1431800"],"award-info":[{"award-number":["23ZR1431800"]}]},{"name":"Shanghai Institute of Pharmaceutical Industry Co. Ltd.","award":["NKLKF2023001"],"award-info":[{"award-number":["NKLKF2023001"]}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2025,7,1]]},"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Immunocytokines targeting immune checkpoints have shown great potential in overcoming resistance to immune checkpoint blockade (ICB), making them a major focus of development in recent years. However, severe dose-limiting toxicity hindered their clinical application. Therefore, it is vital to develop versatile strategies to improve safety and elucidate the underlying mechanisms of resistance reversal for advancing immunocytokine therapy.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A general prodrug platform was established to construct interleukin (IL)-15 and IL-12-based immunocytokine prodrugs (P-T, P-Y, and P-Y-IL12). The efficacy of the masking strategy was validated by in vitro activity assays and in vivo safety evaluations. Antitumor efficacy of P-T was assessed in two murine cold tumor models. A comprehensive immune correlate analysis was conducted in the tumor and tumor-draining lymph node (TDLN) to identify key effector cells responsible for overcoming resistance, followed by further confirmation with egression of T cell blockade, surgical excision of TDLN, and adoptive transfer experiments. Finally, the synergistic antitumor effects of P-T with other ICB or HPK1 inhibitors were investigated.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      P-T or P-Y using steric hindrance from antibody moiety Fab and Fc shields IL-15 activity in circulation, and reactivates it on cleavage by tumor-specific proteases. The universality of this masking strategy is also applicable to IL-12. Compared with prior constructs, P-T and P-Y exhibit prolonged half-life and tumor retention, facilitating sustained intratumoral immune response. P-T demonstrates reduced systemic toxicity but better control of established tumors over the unmasked counterpart. CD44\n                      <jats:sup>+<\/jats:sup>\n                      CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells in TDLNs are identified as critical mediators of P-T\u2019s efficacy: blockade of CD44\n                      <jats:sup>+<\/jats:sup>\n                      CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cell trafficking into the tumor microenvironment (TME) markedly diminishes its antitumor effects. On P-T treatment, CD44\n                      <jats:sup>+<\/jats:sup>\n                      CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells exhibit enhanced proliferation in TDLNs and improved antitumor activity. Furthermore, P-T combined with other immunotherapies enhances antitumor effects by increasing CD44\n                      <jats:sup>+<\/jats:sup>\n                      CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells in TDLNs or promoting their infiltration into the TME.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The Fab and Fc-masked prodrug serves as a universal strategy for next-generation immunocytokines design, effectively addressing their dose-limiting toxicity. Additionally, leveraging immunocytokines to mobilize T cells in TDLNs offers a promising therapy option to overcome resistance to ICB and HPK1 inhibitors.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-011944","type":"journal-article","created":{"date-parts":[[2025,7,31]],"date-time":"2025-07-31T08:45:29Z","timestamp":1753951529000},"page":"e011944","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":8,"title":["Universal Fab-Fc masked cytokine prodrug platform: \u03b1PD-L1\/IL-15 prodrug activates CD44\n                    <sup>+<\/sup>\n                    CD8\n                    <sup>+<\/sup>\n                    T cells in tumor-draining lymph nodes to enhance antitumor immunity"],"prefix":"10.1136","volume":"13","clinical-trial-number":[{"clinical-trial-number":"nct03567889","registry":"10.18810\/clinical-trials-gov"},{"clinical-trial-number":"nct02938299","registry":"10.18810\/clinical-trials-gov"},{"clinical-trial-number":"nct05052268","registry":"10.18810\/clinical-trials-gov"}],"author":[{"given":"Luyao","family":"Song","sequence":"first","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gen","family":"Qi","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenqiang","family":"Shi","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"},{"name":"Department of Cancer Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiongya","family":"Zeng","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Qin","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zejian","family":"Wang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education; School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Li","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Innovative Immunotherapy; 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