Extensive, large-scale single-cell profiling of healthy human blood at different ages is one of the critical pending tasks required to establish a framework for the systematic understanding of human aging. Here, using single-cell RNA/T cell receptor (TCR)/BCR-seq with protein feature barcoding, we profiled 317 samples from 166 healthy individuals aged 25–85 years old. From this, we generated a dataset from ∼2 million cells that described 55 subpopulations of blood immune cells. Twelve subpopulations changed with age, including the accumulation of GZMK⁺CD8⁺ T cells and HLA-DR⁺CD4⁺ T cells. In contrast to other T cell memory subsets, transcriptionally distinct NKG2C⁺GZMB⁻CD8⁺ T cells counterintuitively decreased with age. Furthermore, we found a concerted age-associated increase in type 2/interleukin (IL)4-expressing memory subpopulations across CD4⁺ and CD8⁺ T cell compartments (CCR4⁺CD8⁺ Tcm and Th2 CD4⁺ Tmem), suggesting a systematic functional shift in immune homeostasis with age. Our work provides novel insights into healthy human aging and a comprehensive annotated resource.

对不同年龄的健康人类血液进行广泛、大规模的单细胞分析是建立系统理解人类衰老框架所需的关键悬而未决的任务之一。在这里，使用带有蛋白质特征条形码的单细胞 RNA/T 细胞受体 （TCR）/BCR-seq，我们分析了来自 166 名 25-85 岁健康个体的 317 个样本。由此，我们从大约 200 万个细胞中生成了一个数据集，描述了 55 个血液免疫细胞亚群。12 个亚群随年龄增长而变化，包括 GZMK⁺CD8⁺ T 细胞和 HLA-DR⁺CD4⁺ T 细胞的积累。与其他 T 细胞记忆亚群相比，转录不同的 NKG2C ⁺ GZMB⁻CD8 ⁺ T 细胞随着年龄的增长而反直觉地减少。此外，我们发现 CD4 ⁺ 和 CD8 ⁺ T 细胞区室 （CCR4 ⁺ CD8 ⁺ Tcm 和 Th2 CD4 ⁺ Tmem） 中表达 2 型/白细胞介素 （IL） 4 的记忆亚群与年龄相关的协同增加，表明免疫稳态随着年龄的增长而发生系统性功能转变。我们的工作为健康的人类老龄化提供了新颖的见解和全面的注释资源。