Support data for "Role of adiponectin expressing CD4+ T lymphocytes in regulating immunometabolism"

The immune system and energy metabolism are closely interdependent, with chronic inflammation driving insulin resistance and impaired thermogenesis, while metabolic dysfunction perturbs immune tolerance and promotes inflammation. Here, we investigated the role of adiponectin-expressing CD4 T cells in immunometabolism using mouse models of dietary obesity. Adiponectin-expressing thymocytes localized within thymic nurse cell complexes and were enriched in CD4⁺CD8⁺ double-positive (DP) and CD4 single-positive (SP) populations. Obesity disrupted adiponectin⁺ DP development, reducing CD4 SP frequency and per-cell Adipoq expression. These CD4 SP cells adopted a pro-inflammatory phenotype with diminished Il2 and Cd25 but elevated Il1b, Tnfa, Il6, Il17, and Tgfb, reflecting impaired tolerance and Th17 skewing. Treatment with glyACD, a glycopeptide adiponectin mimetic, expanded adiponectin⁺ thymocytes, restored adiponectin expression at the CD4 SP stage, reduced pro-inflammatory cytokines in metabolic organs, and altered CD100 (Sema4D) distribution, a key T–B interaction molecule. Using conditional CD4 T cell–specific Adipoq knockout and knock-in models, we showed that CD4 T cell–derived adiponectin is essential for glyACD’s metabolic effects. Selective Adipoq expression in CD4 T cells restored insulin sensitivity, lipid metabolism, and energy expenditure, whereas deletion of Adipoq in CD4 T cells abrogated glyACD-induced improvements. Single-cell RNA sequencing revealed that obesity reinforced Sema4D–CD72 interactions between T cells and thymic B cells, while glyACD promoted DP-to-SP maturation and redistributed CD100 interactions. Adiponectin also directly regulated CD100 expression in a cell-autonomous manner. Collectively, our findings identify adiponectin-expressing CD4 T cell as a critical regulator of thymocyte development, T–B interactions, and systemic energy homeostasis.

免疫系统与能量代谢紧密相互依存：慢性炎症可驱动胰岛素抵抗与产热功能受损，而代谢功能障碍则会扰乱免疫耐受并促进炎症发生。本研究利用膳食诱导肥胖的小鼠模型，探究了表达脂联素（adiponectin）的CD4 T细胞在免疫代谢中的作用。表达脂联素的胸腺细胞定位于胸腺抚育细胞复合体（thymic nurse cell complexes）中，并富集于CD4⁺CD8⁺双阳性（double-positive, DP）与CD4单阳性（single-positive, SP）细胞群中。肥胖会破坏脂联素阳性DP细胞的发育，降低CD4 SP细胞的比例以及单个细胞的Adipoq表达水平。这类CD4 SP细胞呈现促炎表型，表现为白细胞介素-2（Il2）与CD25（Cd25）表达降低，而白细胞介素-1β（Il1b）、肿瘤坏死因子-α（Tnfa）、白细胞介素-6（Il6）、白细胞介素-17（Il17）及转化生长因子-β（Tgfb）表达升高，这反映了免疫耐受受损与Th17细胞（T helper 17, Th17）偏态分化。糖肽类脂联素模拟物glyACD可扩增脂联素阳性胸腺细胞，恢复CD4 SP阶段的脂联素表达，降低代谢器官中的促炎细胞因子水平，并改变T-B细胞相互作用关键分子CD100（Sema4D）的分布。通过条件性CD4 T细胞特异性Adipoq敲除与敲入模型，本研究证实CD4 T细胞来源的脂联素是glyACD发挥代谢效应的必需条件。在CD4 T细胞中选择性表达Adipoq可恢复胰岛素敏感性、脂质代谢与能量消耗，而敲除CD4 T细胞中的Adipoq则会消除glyACD诱导的代谢改善效应。单细胞RNA测序（single-cell RNA sequencing）结果显示，肥胖会增强T细胞与胸腺B细胞间的Sema4D–CD72相互作用，而glyACD可促进DP向SP细胞的成熟，并重新分布CD100介导的相互作用。脂联素还能以细胞自主方式直接调控CD100的表达。综上，本研究证实表达脂联素的CD4 T细胞是胸腺细胞发育、T-B细胞相互作用与全身能量稳态的关键调控因子。