{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T00:23:36Z","timestamp":1783470216446,"version":"3.55.0"},"reference-count":54,"publisher":"BMJ","issue":"9","license":[{"start":{"date-parts":[[2025,9,15]],"date-time":"2025-09-15T00:00:00Z","timestamp":1757894400000},"content-version":"unspecified","delay-in-days":14,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100018537","name":"National Science and Technology Major Project of the Ministry of Science and Technology of China","doi-asserted-by":"crossref","award":["2023YFC2508501"],"award-info":[{"award-number":["2023YFC2508501"]}],"id":[{"id":"10.13039\/501100018537","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100018537","name":"National Science and Technology Major Project of the Ministry of Science and Technology of China","doi-asserted-by":"crossref","award":["2023YFC2508505"],"award-info":[{"award-number":["2023YFC2508505"]}],"id":[{"id":"10.13039\/501100018537","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82250710684"],"award-info":[{"award-number":["82250710684"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2025,8,29]]},"published-print":{"date-parts":[[2025,9]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>The strong regulatory T cell (Treg) inhibitory activity and dysfunctional cytotoxic T lymphocytes (CTLs) represent major barriers to effective antitumor immunity, particularly in late-stage cancer. Multiple anti-OX40 (aOX40) agonistic antibodies have been developed but exhibit limited antitumor efficacy. Interleukin-2 (IL-2) effectivity expands CTLs but has severe side effects.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We construct an aOX40-mIL2-Fc bispecific antibody through Fab physical blocking and attenuated IL-2 with R\u03b2 reducing N88D mutation. We also produced aOX40-Fc and IL-2\/aOX40-Fc as a comparison using the 293F expression system. Single-cell and flow cytometry were used to analyze the change of T-cell subsets in the tumor microenvironment (TME). Mouse tumor models were used to assess the antitumor efficacy of aOX40-mIL2-Fc by tumor growth and survival, and toxicity by body weight loss, inflammatory cytokine production, and natural killer (NK) cell proliferation in the blood. The tumor-bearing mice were randomly assigned, and the average size was similar among various groups.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      aOX40-mIL2-Fc bispecific antibody-cytokine exhibited a synergistic therapeutic effect with limited toxicity, outperforming IL-2-Fc or aOX40 alone treatment, and conferring resistance to tumor rechallenge. On cellular mechanisms, aOX40-mIL2-Fc treatment showed great Treg depletion and increased both stem-like and effector functional terminal CD8\n                      <jats:sup>+<\/jats:sup>\n                      T cells in the TME, while avoiding NK cells expansion in the periphery. Furthermore, this bispecific antibody remarkably improved the anti-programmed death-ligand 1 (PD-L1) therapeutic effect.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our study unveils a novel approach to IL-2 design that addresses several critical shortcomings of existing strategies and elucidates the cellular mechanisms underlying aOX40-mIL2-Fc therapy. Meanwhile, combining aOX40-mIL2-Fc with PD-L1 blockade represents a strategic approach to enhance tumor control and overcome resistance to immune checkpoint blockade therapies synergistically.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-011638","type":"journal-article","created":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T02:46:05Z","timestamp":1757990765000},"page":"e011638","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":5,"title":["Dual targeting OX40 and IL-2 receptor enhances antitumor activity through tumor-infiltrating Treg depletion and CD8\n                    <sup>+<\/sup>\n                    T-cell proliferation"],"prefix":"10.1136","volume":"13","author":[{"given":"Shuaishuai","family":"Cao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Molecular Oncology and School of Basic Medical Sciences, Tsinghua University, Beijing, 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