Negin Farzad, Archibald Enninful, Yao Lu, Fabio Parisi, Anthony A. Fung, Yumi Kwon, Yajuan Li, Margaux Labrosse, Mingyu Yang, Francesco Strino, Liang Chen, Junchen Yang, Mei Zhong, Fu Gao, Bo Tao, Joseph Cunningham, Zhiliang Bai, Haikuo Li, fang wang, Michael Stankewich, Dongjoo Kim, Mingze Dong, Lisa Bramer, Keyi Li, Meera Bhat, Evan J. Loe, Joseph Craft, Ljiljana Pasa-Tolic, Stephanie Halene, Lingyan Shi, Yuval Kluger, Mina L. Xu, Rong Fan
Immunosenescence is a hallmark of human aging and contributes to age-related immune decline, yet the development of senescence-associated phenotypes in the human lymphoid organs remains poorly understood. Here, we integrate single-cell and spatial multi-omics to systematically characterize age-related senescence in human lymph nodes (LNs) across the lifespan. Spatial proteomic profiling of 99 LN sections from 51 donors (18–86 years) using high-plex immunofluorescence (∼20 million cells) mapped senescence markers (p16, p21, HMGB1, and γ-H2AX) at single-cell resolution, revealing diverse senescent-like cell types ("senotypes") and a stepwise shift from extrafollicular to germinal-center localization with age. In aged LNs, germinal-center B cells exhibit focal accumulation of senescence-associated programs, accompanied by impaired functional signatures, metabolic remodeling, and altered regulatory networks. These findings define a spatially organized landscape of immunosenescence in human lymphoid tissue and highlight germinal-center B cells as a key locus of age-associated immune dysfunction.