Transcriptomic profiling of human macrophages following adjuvant stimulation. Transcriptomic profiling of human macrophages following adjuvant stimulation

Trained immunity is the phenomenon whereby innate immune cells such as monocytes or macrophages undergo functional reprogramming after exposure to certain microbial components, altering their responses to future exposures. Since trained immunity was first discovered, and has been commonly studied with the BCG vaccine, studies confirming trained immunity have often focused on human PBMCs. Thus, to study the mechanism of the vaccine in a well-established trained immunity model, we used human macrophages differentiated from monocytes isolated from PBMCs. The macrophages were expose to A + M + MA for 24 h, and rested two or five days prior to exposure to bacteria, as an ex vivo model with a five-day rest period has been used in numerous studies to confirm trained immunity. Overall design: PBMCs were obtained from healthy donors, and stimulated with the vaccine for 24 hours and rested for 2 or 5 days. Expeirment was repeated 3 times to obtain three biological replicates.

训练免疫（trained immunity）是指单核细胞、巨噬细胞等固有免疫细胞在接触特定微生物组分后发生功能重编程，进而改变其对后续暴露的应答反应的现象。自训练免疫被首次发现以来，卡介苗（BCG vaccine）是其最常用的研究模型，因此相关验证研究多以人外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）为研究对象。为了在成熟的训练免疫模型中探究该疫苗的作用机制，我们采用了从PBMCs中分离的单核细胞分化得到的人源巨噬细胞。将巨噬细胞用A+M+MA处理24小时，随后静置2或5天再接触细菌；鉴于静置5天的离体模型已在多项验证训练免疫的研究中被广泛应用，本研究采用该实验设置。整体实验设计如下：从健康供者体内分离PBMCs，用该疫苗刺激24小时后静置2或5天；本实验重复3次，以获得3次生物学重复。