2026-10-10T16:06:33Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33940402 2026-09-18T18:02:14Z category_4 category_12 category_13 category_14 category_46 category_61 category_64 category_132 portal_5 item_type_3 month_year_09_2026

Supporting dataset. Ravear Zhiqiang Wang (25073635) Fang Yang (123514) Jianhua Sui (53799) Biochemistry Cell Biology Genetics Molecular Biology Immunology Developmental Biology Cancer Infectious Diseases sup >+</ sup negative immune regulation functional assays revealed complex regulatory network three knockout models single component exerting additive phenotype compared various cancer cells knockout mice showed div >< p detailed molecular characterization cd28 downstream signaling cell signaling within uncovers additional gal h4 deficiency produced b7 family members 9 binding partners h4 binding partner h4 – gal h4 inhibits gal knockout mice binding partner detectable phenotype mice resulted cell responses cell death cell activity vivo </ vitro </ tumor responses tumor immunity study provides splenic cd4 offering insights mechanisms underlying levels comparable induced activation increased proportion igc domain glycosylation within findings indicate dominant effect dependent interaction broadly expressed b7 family also bind 9 single 9 deficiency <div>B7-H4, a member of the B7 family, is broadly expressed on various cancer cells and has been implicated in negative immune regulation, particularly in suppressing anti-tumor immunity. However, its receptor and the mechanisms underlying its immunosuppression remain poorly understood. Here, we identify Galectin-9 (Gal-9) as a binding partner of B7-H4 and investigate its role in modulating T cell responses. We show that glycosylation within the IgC domain of B7-H4 is required for Gal-9 binding, while the N-terminal carbohydrate recognition domain (N-CRD) of Gal-9—specifically residue R65—is essential for its binding with B7-H4. In addition, several other B7 family members (B7.1, B7.2, B7-H2, and B7-DC) and immune cell surface receptors (CD28, 2B4, CD226, and SLAMF1) also bind to Gal-9 at levels comparable to those observed with B7-H4 or TIM-3. In vitro functional assays revealed that B7-H4 inhibits Gal-9-induced activation of CD28 downstream signaling and reduces Gal-9-mediated T cell death. In vivo, Gal-9 deficiency in mice resulted in an increased proportion of splenic CD4+ T cells, whereas B7-H4 deficiency produced no detectable phenotype. Moreover, B7-H4 and Gal-9 double-knockout mice showed no additive phenotype compared with Gal-9 single-knockout mice, and tumor growth following tumor cell challenge was unaffected in all three knockout models. Collectively, these findings indicate that B7-H4, Gal-9, other B7 family members, and T cell surface immune receptors form a complex regulatory network that modulates T cell activity and anti-tumor responses, with no single component exerting a dominant effect. This study provides a detailed molecular characterization of the B7-H4–Gal-9 interaction and uncovers additional Gal-9 binding partners, offering insights into the finely tuned immune regulation mediated by the B7 family.</div> 2026-09-18T18:02:03Z Dataset Dataset 10.1371/journal.pone.0355964.s008 https://figshare.com/articles/dataset/_p_Supporting_dataset_p_/33940402 CC BY 4.0