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.